CN117769433A

CN117769433A - 冠状病毒疫苗制剂

Info

Publication number: CN117769433A
Application number: CN202280036535.7A
Authority: CN
Inventors: G·史密斯; M·J·马萨尔; 田京慧
Original assignee: Novavax Inc
Current assignee: Novavax Inc
Priority date: 2021-03-22
Filing date: 2022-03-18
Publication date: 2024-03-26

Abstract

本申请公开了适合于在疫苗中使用的冠状病毒刺突(S)蛋白和包含所述冠状病毒刺突蛋白的纳米颗粒。所述纳米颗粒呈递来自包围洗涤剂核心并与之缔合的病原体的抗原，从而产生增强的稳定性和良好的免疫原性。还公开了所述疫苗和纳米颗粒的剂量、制剂和用于制备所述疫苗和纳米颗粒的方法。

Description

冠状病毒疫苗制剂

相关申请的交叉引用

本申请要求以下申请的优先权：2021年3月22日提交的美国临时申请号63/164,487；2021年4月19日提交的美国临时申请号63/176,825；2021年4月20日提交的美国临时申请号63/177,059；2021年6月2日提交的美国临时申请号63/195,986；2021年11月17日提交的美国临时申请号63/280,395；2021年12月16日提交的美国临时申请号63/290,439；2021年12月21日提交的美国临时申请号63/292,196；以及2021年12月23日提交的美国临时申请号63/293,468。将每一个上述申请的内容通过引用以其整体并入本申请申请。本申请还将国际公开号2021/0154812和美国专利号10,953,089通过引用以其整体并入本申请。

电子提交的文本申请件的说明

将与本申请一起以电子方式提交的文本申请件的内容通过引用以其整体并入本申请申请：序列表的计算机可读格式拷贝(文件名：NOVV_092_01WO_SeqList_ST25.txt，记录日期：2022年3月9日，文件大小：约974千字节)。

技术领域

本申请公开文本总体上涉及非天然存在的冠状病毒(CoV)刺突(S)多肽以及包含所述多肽的纳米颗粒和疫苗，其可用于刺激免疫反应。所述纳米颗粒提供了任选地与洗涤剂核心缔合的抗原(例如，糖蛋白抗原)，并且通常使用重组方法产生。所述纳米颗粒具有改善的稳定性和增强的表位呈递。本申请公开文本还提供了含有所述纳米颗粒的组合物、用于产生所述组合物的方法以及刺激免疫反应的方法。

背景技术

感染性疾病仍然是遍及全世界的问题。尽管在开发针对一些病原体的疫苗方面已取得进展，但许多病原体仍对人类健康构成威胁。仅在美国，突发急性呼吸系统综合征冠状病2(SARS-CoV-2)的爆发就感染了超过7900万人，且至少有960,000人死亡。在全球范围内，死亡人数已超过600万。SARS-CoV-2冠状病毒与在过去17年中已导致数百人死亡的严重急性呼吸系统综合征冠状病毒(SARS-CoV)和中东呼吸系统综合征冠状病毒(MERS-CoV)属于同一病毒家族。SARS-CoV-2导致疾病COVID-19。世界卫生组织于2020年3月将COVID-19分类为全球大流行病。这种大流行病仍在持续。SARS-CoV-2的变体的出现阻碍了控制它所做的努力。

期望开发出预防由SARS-CoV-2及其变体引起的危及生命的感染性疾病或减轻所述疾病的严重程度的疫苗。然而，由于病原体的逃避机制非常复杂并且难以使疫苗稳定，因此人类疫苗的开发仍然具有挑战性。最佳地，疫苗必须同时诱导阻断或中和感染原的抗体，并且在各种环境(包括不能制冷的环境)中保持稳定。

发明内容

本申请公开文本提供了适用于诱导针对SARS-CoV-2和SARS-CoV-2变体的免疫反应的非天然存在的CoV S多肽。本申请公开文本还提供了含有所述糖蛋白的纳米颗粒以及刺激免疫反应的方法。

本申请公开文本还提供了适用于诱导针对多种冠状病毒的免疫反应的CoV S多肽，所述冠状病毒包括SARS-CoV-2及其变体、中东呼吸系统综合征(MERS)和严重急性呼吸系统综合征(SARS)。

本申请提供了CoV S多肽，所述多肽包含：

(i)具有灭活的弗林蛋白酶切割位点的S1亚基，其中所述S1亚基包含N末端结构域(NTD)、受体结合结构域(RBD)、亚结构域1和2(SD1/2)，其中所述灭活的弗林蛋白酶切割位点具有QQAQ的氨基酸序列；

其中所述NTD任选地包含选自以下的一个或多个修饰：

(a)选自氨基酸56、57、131、132、144、145、228、229、230、231、234、235、236、237、238、239、240及其组合的一个或多个氨基酸的缺失；

(b)在氨基酸132之后1、2、3或4个氨基酸的插入；以及

(c)选自氨基酸5、6、7、13、39、51、53、54、56、57、62、63、67、82、125、129、131、132、133、139、143、144、145、177、200、201、202、209、229、233、240、245及其组合的一个或多个氨基酸的突变；

其中所述RBD任选地包含选自氨基酸333、404、419、426、439、440、464、465、471、477、481、488及其组合的一个或多个氨基酸的突变；

其中所述SD1/2结构域任选地包含选自557、600、601、642、664、668及其组合的一个或多个氨基酸的突变；以及

(ii)S2亚基，其中氨基酸973和974是脯氨酸，其中所述S2亚基任选地包含选自以下的一个或多个修饰：

(a)从676-685、676-702、702-711、775-793、806-815及其组合中的一个或多个氨基酸的缺失；

(b)选自688、703、846、875、937、969、973、974、1014、1058、1105和1163及其组合的一个或多个氨基酸的突变；以及

(c)从TMCT中的一个或多个氨基酸的缺失；其中CoV S糖蛋白的氨基酸关于具有SEQ ID NO:2的序列的多肽进行编号。

在实施方案中，所述冠状病毒S糖蛋白包含氨基酸676-685的缺失。在实施方案中，所述冠状病毒S糖蛋白包含氨基酸702-711的缺失。在实施方案中，所述冠状病毒S糖蛋白包含氨基酸806-815的缺失。在实施方案中，所述冠状病毒S糖蛋白包含氨基酸775-793的缺失。在实施方案中，所述冠状病毒S糖蛋白包含NTD的氨基酸1-292的缺失。在实施方案中，所述冠状病毒S糖蛋白包含TMCT的氨基酸1201-1260的缺失。在实施方案中，所述冠状病毒S糖蛋白包含以下氨基酸序列或由以下氨基酸序列组成：选自SEQ ID NO:85-89、105、106和112-115、164-168的氨基酸序列或者与SEQ ID NO:85-89、105、106和112-115、164-168中的任一个具有至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、至少99％或100％同一性的氨基酸序列。在实施方案中，所述冠状病毒S糖蛋白包含信号肽，任选地其中所述信号肽包含SEQ ID NO:5或SEQ ID NO:117的氨基酸序列。在实施方案中，所述冠状病毒S糖蛋白包含C末端融合蛋白。在一些实施方案中，所述C末端融合蛋白是六聚组氨酸标签。在一些实施方案中，所述C末端融合蛋白是折叠子。在一些实施方案中，所述折叠子具有对应于SEQ ID NO:68的氨基酸序列。在一些实施方案中，所述冠状病毒S糖蛋白具有如下ΔHcal，其是野生型CoV S糖蛋白(SEQ ID NO:2)的ΔHcal的至少2倍。在实施方案中，本申请提供了具有S2亚基、NTD、RBD和SD1/2的冠状病毒S糖蛋白，其与具有SEQ ID NO:2的氨基酸序列的CoV S糖蛋白的相应亚基或结构域至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、至少99％或至少99.5％相同。

本申请提供了编码本申请所述的CoV S糖蛋白的分离核酸。

本申请提供了包含编码本申请所述的CoV S糖蛋白的分离核酸的载体。

本申请提供了包含本申请所述的CoV S糖蛋白的纳米颗粒。在一些实施方案中，所述纳米颗粒的Zavg直径在约20nm与约35nm之间。在一些实施方案中，所述纳米颗粒的多分散性指数为约0.2至约0.45。本申请提供了一种表达本申请所述的CoV S糖蛋白的细胞。

本申请提供了一种包含含有本申请所述的CoV S糖蛋白的纳米颗粒的疫苗组合物。在一些实施方案中，所述疫苗组合物包含佐剂。在一些实施方案中，所述佐剂包含至少两种iscom颗粒，其中：第一iscom颗粒包含皂树(Quillaja Saponaria Molina)的级分A而不包含皂树的级分C；并且第二iscom颗粒包含皂树的级分C而不包含皂树的级分A。在一些实施方案中，皂树的级分A和皂树的级分C分别占在所述佐剂中皂树的级分A和皂树的级分C的重量之和的按重量计约85％和按重量计约15％。在实施方案中，皂树的级分A和皂树的级分C分别占在所述佐剂中皂树的级分A和皂树的级分C的重量之和的按重量计约92％和按重量计约8％。在一些实施方案中，所述疫苗组合物包含约50μg佐剂。

本申请提供了一种在受试者中刺激针对SARS-CoV-2的免疫反应的方法，所述方法包括施用本申请所述的疫苗组合物。在实施方案中，向所述受试者在第0天施用第一个剂量，并且在第21天施用加强剂量。在实施方案中，向所述受试者施用约3μg至约25μg的冠状病毒S糖蛋白。在实施方案中，向所述受试者施用约5μg的冠状病毒S糖蛋白。在实施方案中，将所述疫苗组合物肌内施用。在实施方案中，施用单剂量的所述疫苗组合物。在实施方案中，施用多个剂量的所述疫苗组合物。在实施方案中，将所述疫苗组合物与流感糖蛋白共同施用。

本申请提供了一种免疫原性组合物，所述免疫原性组合物包含：(i)含有本申请所述的CoV S糖蛋白和非离子型洗涤剂核心的纳米颗粒；(ii)医药上可接受的缓冲液；以及(iii)皂苷佐剂。在实施方案中，所述免疫原性组合物包含约3μg至约25μg的CoV S糖蛋白。在实施方案中，所述免疫原性组合物包含约5μg的CoV S糖蛋白。在实施方案中，所述皂苷佐剂包含至少两种iscom颗粒，其中：第一iscom颗粒包含皂树的级分A而不包含皂树的级分C；并且第二iscom颗粒包含皂树的级分C而不包含皂树的级分A。在实施方案中，分别地，皂树的级分A占在所述佐剂中皂树的级分A和皂树的级分C的重量之和的按重量计50％-96％，并且皂树的级分C占剩余部分。在实施方案中，皂树的级分A和皂树的级分C分别占在所述佐剂中皂树的级分A和皂树的级分C的重量之和的按重量计约85％和按重量计约15％。在实施方案中，所述免疫原性组合物包含约50μg的皂苷佐剂。在实施方案中，所述非离子型洗涤剂选自聚山梨醇酯20(PS20)、聚山梨醇酯40(PS40)、聚山梨醇酯60(PS60)、聚山梨醇酯65(PS65)和聚山梨醇酯80(PS80)。

在实施方案中，本申请提供了一种在受试者中刺激针对SARS-CoV-2或异源SARS-CoV-2毒株的免疫反应的方法，所述方法包括施用本申请提供的免疫原性组合物或疫苗组合物。在实施方案中，所述方法包括施用约3μg至约25μg的CoV S糖蛋白。在实施方案中，所述方法包括施用约5μg的CoV S糖蛋白。在实施方案中，所述皂苷佐剂包含至少两种iscom颗粒，其中：第一iscom颗粒包含皂树的级分A而不包含皂树的级分C；并且第二iscom颗粒包含皂树的级分C而不包含皂树的级分A。在实施方案中，分别地，皂树的级分A占在所述佐剂中皂树的级分A和皂树的级分C的重量之和的按重量计50％-96％，并且皂树的级分C占剩余部分。在实施方案中，皂树的级分A和皂树的级分C分别占皂树的级分A和皂树的级分C的重量之和的按重量计约85％和按重量计约15％。在实施方案中，所述方法包括施用约50μg的皂苷佐剂。在实施方案中，所述非离子型洗涤剂选自聚山梨醇酯20(PS20)、聚山梨醇酯40(PS40)、聚山梨醇酯60(PS60)、聚山梨醇酯65(PS65)和聚山梨醇酯80(PS80)。在实施方案中，向所述受试者在第0天施用第一个剂量，并且在第21天施用加强剂量。在实施方案中，施用单剂量的所述免疫原性组合物。在实施方案中，所述方法包括施用第二免疫原性组合物。在实施方案中，所述第二免疫原性组合物包含编码SARS-CoV-2刺突糖蛋白的mRNA、编码SARS-CoV-2刺突糖蛋白的质粒DNA、编码SARS-CoV-2刺突糖蛋白的病毒载体或灭活的SARS-CoV-2病毒。在实施方案中，所述异源SARS-CoV-2毒株选自B.1.1.7SARS-CoV-2毒株、B.1.351SARS-CoV-2毒株、P.1SARS-CoV-2毒株、B.1.617.2SARS-CoV-2毒株、B.1.525SARS-CoV-2毒株、B.1.526SARS-CoV-2毒株、B.1.617.1SARS-CoV-2毒株、C.37SARS-CoV-2毒株、B.1.621SARS-CoV-2毒株和Cal.20C SARS-CoV-2毒株。在实施方案中，所述免疫原性组合物用于预防冠状病毒病-19(COVID-19)的功效为至少约50％、至少约55％、至少约60％、至少约65％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％、至少约91％、至少约92％、至少约93％、至少约94％、至少约95％、至少约96％、至少约97％、至少约98％、至少约99％或约100％，所述功效在施用所述免疫原性组合物后持续长达约2个月、长达约2.5个月、长达约3个月、长达约3.5个月、长达约4个月、长达约4.5个月、长达约5个月、长达约5.5个月、长达约6个月、长达约6.5个月、长达约7个月、长达约7.5个月、长达约8个月、长达约8.5个月、长达约9个月、长达约9.5个月、长达约10个月、长达约10.5个月、长达约11个月、长达约11.5个月或长达约12个月。在实施方案中，所述免疫原性组合物用于预防冠状病毒病-19(COVID-19)的功效为从约50％至约99％、从约50％至约95％、从约50％至约90％、从约50％至约85％、从约50％至约80％、从约60％至约99％、从约60％至约95％、从约60％至约90％、从约60％至约85％、从约60％至约80％、从约40％至约99％、从约40％至约95％、从约40％至约90％、从约40％至约85％、从约40％至约80％、从约40％至约75％、从约40％至约70％、从约40％至约65％、从约40％至约55％或从约40％至约50％，所述功效在施用所述免疫原性组合物后持续长达约2个月、长达约2.5个月、长达约3个月、长达约3.5个月、长达约4个月、长达约4.5个月、长达约5个月、长达约5.5个月、长达约6个月、长达约6.5个月、长达约7个月、长达约7.5个月、长达约8个月、长达约8.5个月、长达约9个月、长达约9.5个月、长达约10个月、长达约10.5个月、长达约11个月、长达约11.5个月或长达约12个月。在实施方案中，所述COVID-19是轻度COVID-19。在实施方案中，所述COVID-19是中度COVID-19。在实施方案中，所述COVID-19是重度COVID-19。在实施方案中，本申请提供了一种诱导针对异源SARS-CoV-2毒株的保护性免疫反应的方法，所述方法包括向受试者施用包含具有SEQ IDNO:87的氨基酸序列的冠状病毒S(CoV S)糖蛋白和非离子型洗涤剂核心的纳米颗粒、医药上可接受的缓冲液以及(iii)皂苷佐剂，其中所述异源SARS-CoV-2毒株具有与SEQ ID NO:1的SARS-CoV-2糖蛋白相比含有约1至约60个修饰的SARS-CoV-2S糖蛋白。在实施方案中，所述异源SARS-CoV-2毒株具有与SEQ ID NO:1的SARS-CoV-2糖蛋白相比含有约1至约20个修饰、约1至约10个修饰、约10至约20个修饰、10至约30个修饰、约10至约40个修饰、10至约50个修饰、10至约60个修饰、20至约60个修饰、20至约50个修饰、约20至约40个修饰、约5至约15个修饰或约5至约10个修饰的SARS-CoV-2S糖蛋白。

附图说明

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图1显示了SARS-CoV-2刺突(S)蛋白的野生型氨基酸序列(SEQ ID NO:1)的示意图。弗林蛋白酶切割位点RRAR(SEQ ID NO:6)以粗体突出显示，并且信号肽加下划线。

图2显示了SARS-CoV-2S多肽的一级结构，其具有无活性弗林蛋白酶切割位点、融合肽缺失以及K986P和V987P突变。结构域位置关于来自含有信号肽的SARS-CoV-2的野生型CoV S多肽的氨基酸序列(SEQ ID NO:1)进行编号。

图3显示了BV2378 CoV S多肽的一级结构，其具有无活性弗林蛋白酶切割位点、氨基酸819-828的融合肽缺失以及K986P和V987P突变。结构域位置关于来自含有信号肽的SARS-CoV-2的野生型CoV S多肽的氨基酸序列(SEQ ID NO:1)进行编号。

图4显示了CoV S多肽BV2364、BV2365、BV2366、BV2367、BV2368、BV2369、BV2373、BV2374和BV2375的纯化。数据揭示具有含有QQAQ的氨基酸序列(SEQ ID NO:7)的无活性弗林蛋白酶切割位点的BV2365(SEQ ID NO:4)和BV2373(SEQ ID NO:87)被表达为单链(S0)。相比之下，含有完整弗林蛋白酶切割位点的CoV S多肽(例如BV2364、BV2366和BV2374)被切割，如通过切割产物S2的存在所证实的。

图5通过生物膜层干涉测量术显示，CoV S多肽BV2361、BV2365、BV2369、BV2365、BV2373和BV2374与人血管紧张素转化酶2前体(hACE2)结合。

图6通过生物膜层干涉测量术显示，来自SARS-CoV-2的BV2361不结合MERS-CoV受体二肽基肽酶IV(DPP4)，并且MERS S蛋白不与人血管紧张素转化酶2前体(hACE2)结合。

图7通过酶联免疫吸附测定(ELISA)显示，BV2361与hACE2结合。

图8显示了BV2373 CoV S多肽的一级结构和对弗林蛋白酶切割位点的修饰K986P和V987P。

图9显示了野生型CoV S多肽以及CoV S多肽BV2365和BV2373的纯化。

图10显示了覆盖在SARS-CoV-2刺突蛋白(EMB ID：21374)的冷冻电子显微术(cryoEM)结构上的BV2373 CoV S多肽的cryoEM结构。

图11A-图11F显示CoV S刺突多肽BV2365和BV2373与hACE2结合。生物膜层干涉测量术揭示，BV2365(图11B)和BV2373(图11C)以与野生型CoV S多肽(图11A)类似的解离动力学与hACE2结合。ELISA表明，野生型CoV S多肽(图11D)和BV2365(图11E)以类似的亲和力与hACE2结合，而BV2373以更高的亲和力与hACE2结合(图11F)。

图12A-图12B显示了胁迫条件(例如温度、两个冷冻/解冻循环、氧化、搅拌和pH极端值)对CoV S多肽BV2373(图12A)和BV2365(图12B)与hACE2的结合的影响。

图13A-图13B显示了在用两个剂量(图13A)和一个剂量(图13B)的0.1μg至10μg的BV2373、加或不加级分A和级分C iscom基质(即，MATRIX-M^TM)免疫小鼠后13天、21天和28天的抗CoV S多肽IgG滴度。

图14显示了在用一个剂量或两个剂量的0.1μg至10μg的BV2373、加或不加MATRIX-M^TM免疫的小鼠中阻断hACE2相互作用的抗体的诱导。

图15显示了在用一个剂量或两个剂量的0.1μg至10μg的BV2373、加或不加MATRIX-M^TM免疫的小鼠中检测到的病毒中和抗体。

图16显示了用单剂量的BV2373或两个剂量的BV2373间隔14天、加或不加MATRIX-M^TM免疫的Ad/CMV/hACE2小鼠的肺中的病毒载量(SARS-CoV-2)。

图17A-图17C显示了用BV2373免疫后小鼠展现出的体重减轻。图17A显示了用单一0.01μg、0.1μg、1μg或10μg的BV2373+MATRIX-M^TM免疫对体重减轻的影响。图17B显示了用两个剂量的BV2373(0.01μg、0.1μg、1μg)+MATRIX-M^TM免疫对体重减轻的影响。图17C显示了在存在或不存在MATRIX-M^TM的情况下用两个剂量的BV2373(10μg)免疫对体重减轻的影响。

图18A-图18B显示了BV2373在感染SARS-CoV-2后四天(图18A)或七天(图18B)对小鼠肺组织病理学的影响。

图19显示了与在存在MATRIX-M^TM的情况下用BV2373免疫的小鼠相比，在不存在佐剂的情况下用BV2373免疫的小鼠的脾脏中在体外刺激后IFN-γ分泌性细胞的数量。

图20A-图20E显示了在存在或不存在MATRIX-M^TM的情况下用BV2373免疫的小鼠的脾脏中细胞因子分泌性CD4+T细胞的频率。图20A显示了IFN-γ分泌性CD4+T细胞的频率。图20B显示了TNF-α分泌性CD4+T细胞的频率。图20C显示了IL-2分泌性CD4+T细胞的频率。图20D显示了分泌选自IFN-γ、TNF-α和IL-2的两种细胞因子的CD4+T细胞的频率。图20E显示了表达IFN-γ、TNF-α和IL-2的CD4+T细胞的频率。

图21A-图21E显示了在存在或不存在MATRIX-M^TM的情况下用BV2373免疫的小鼠的脾脏中细胞因子分泌性CD8⁺T细胞的频率。图21A显示了IFN-γ分泌性CD8⁺T细胞的频率。图21B显示了TNF-α分泌性CD8⁺T细胞的频率。图21C显示了IL-2分泌性CD8⁺T细胞的频率。图20D显示了分泌选自IFN-γ、TNF-α和IL-2的两种细胞因子的CD8⁺T细胞的频率。图21E显示了表达IFN-γ、TNF-α和IL-2的CD8⁺T细胞的频率。

图22展示了在存在或不存在MATRIX-M^TM的情况下用BV2373免疫的小鼠的脾脏中表达选自IFN-γ、TNF-α和IL-2的一种(单一)、两种(双重)或三种(三重)细胞因子的CD4⁺或CD8⁺细胞的频率。

图23A-图23C展示了在存在或不存在MATRIX-M^TM的情况下用BV2373免疫对CD4⁺T细胞分泌2型细胞因子的影响。图23A显示了IL-4分泌性细胞的频率。图23B显示了IL-5分泌性CD4⁺细胞的频率。图23C显示了IFN-γ分泌性与IL-4分泌性CD4⁺T细胞的比率。

图24A-图24B通过评估CD4⁺滤泡辅助性T细胞(TFH)的存在展示了在存在或不存在MATRIX-M^TM的情况下用BV2373免疫小鼠对生发中心形成的影响。图24A显示了脾脏中CD4⁺滤泡辅助性T细胞的频率，并且图24B显示了CD4⁺滤泡辅助性T细胞的表型(例如CD4⁺CXCR5⁺PD-1⁺)。

图25A-图25B通过评估生发中心(GC)B细胞的存在展示了在存在或不存在MATRIX-M^TM的情况下用BV2373免疫小鼠对生发中心形成的影响。图25A显示了脾脏中GC B细胞的频率，并且图25B揭示了CD4⁺滤泡辅助性T细胞的表型(例如CD19⁺GL7⁺CD-95⁺)。

图26A-图26C显示了在存在或不存在MATRIX-M^TM的情况下用BV2373免疫对橄榄狒狒的抗体反应的影响。图26A显示了用BV2373免疫后狒狒中的抗SARS-CoV-2S多肽IgG滴度。图26B显示了在存在MATRIX-M^TM的情况下用5μg或25μg的BV2373单次免疫后在狒狒中hACE2受体阻断抗体的存在。图26C显示了用BV2373和MATRIX-M^TM单次免疫后病毒中和抗体的滴度。

图27显示了用BV2373免疫后橄榄狒狒中抗SARS-CoV-2S多肽IgG与中和抗体滴度之间的显著相关性。

图28显示了在存在或不存在MATRIX-M^TM的情况下用BV2373免疫的橄榄狒狒的外周血单核细胞(PBMC)中IFN-γ分泌性细胞的频率。

图29A-图29E显示了在存在或不存在MATRIX-M^TM的情况下用BV2373免疫的橄榄狒狒的PBMC中细胞因子分泌性CD4+T细胞的频率。图29A显示了IFN-γ分泌性CD4+T细胞的频率。图29B显示了IL-2分泌性CD4+T细胞的频率。图29C显示了TNF-α分泌性CD4+T细胞的频率。图29D显示了分泌选自IFN-γ、TNF-α和IL-2的两种细胞因子的CD4+T细胞的频率。图29E显示了表达IFN-γ、TNF-α和IL-2的CD4+T细胞的频率。

图30显示了冠状病毒刺突(S)蛋白(SEQ ID NO:109)(BV2384)的示意图。弗林蛋白酶切割位点GSAS(SEQ ID NO:97)加单下划线，并且K986P和V987P突变加双下划线。

图31显示了冠状病毒刺突(S)蛋白(SEQ ID NO:86)(BV2373)的示意图。弗林蛋白酶切割位点QQAQ(SEQ ID NO:7)加单下划线，并且K986P和V987P突变加双下划线。

图32显示了CoV S多肽BV2373(SEQ ID NO:87)和BV2384(SEQ ID NO:109)的纯化。

图33显示了纯化后BV2384(SEQ ID NO:109)纯度的扫描光密度测定图。

图34显示了纯化后BV2373(SEQ ID NO:87)纯度的扫描光密度测定图。

图35A-图35B展示了响应于BV2373和MATRIX-M^TM的施用抗S抗体(图35A)和中和抗体(图35B)的诱导。向食蟹猴施用一个或两个剂量(第0天和第21天)的2.5μg、5μg或25μg的BV2373加25μg或50μg MATRIX-M^TM佐剂。对照既未接受BV2373又未接受MATRIX-M^TM。在第21天和第33天测量抗体。

图36A-图36B展示了本申请所公开的疫苗制剂对SARS-CoV-2病毒复制的减少，如在食蟹猴的支气管肺泡灌洗液(BAL)中所评估的。如图所示，向食蟹猴施用BV2373和MATRIX-M^TM。受试者在第0天进行免疫，并且在用两个剂量的组中在第0天和第21天进行免疫。第37天用1x10⁴ pfu SARS-CoV-2病毒攻击受试动物。在用感染性病毒攻击后2天和4天(d2pi和d4pi)，在支气管灌洗液(BAL)中评估病毒RNA(图36A，对应于存在的总RNA)和病毒亚基因组RNA(图36B，对应于复制病毒)水平。大多数受试者未显示出病毒RNA。在第2天，在一些受试者中测量到少量RNA。到第4天，除了接受2.5μg最低剂量的两名受试者以外，都未测量到RNA。除了同样接受最低剂量的1名受试者以外，在2天或4天均未检测到亚基因组RNA。

图37A-图37B展示了本申请所公开的疫苗制剂对SARS-CoV-2病毒复制的减少，如在食蟹猴的鼻拭子中所评估的。如图所示，向食蟹猴施用BV2373加MATRIX-M^TM。受试者在第0天进行免疫，并且在用两个剂量的组中在第0天和第21天进行免疫。第37天用1x10⁴SARS-CoV-2病毒攻击受试动物。在感染后2天和4天(d2pi和d4pi)通过鼻拭子评估病毒RNA(图37A)和病毒亚基因组(sg)RNA(图37B)。大多数受试者未显示出病毒RNA。在第2天和第4天，在一些受试者中测量到少量RNA。在2天或4天均未检测到亚基因组RNA。受试者在第0天进行免疫，并且在用两个剂量的组中在第0天和第21天进行免疫。这些数据表明，疫苗将鼻总病毒RNA降低100-1000倍，并将sgRNA降低至不可检测的水平，并且确认对疫苗的免疫反应将阻断病毒复制并防止病毒传播。

图38A-图38B显示了用一个剂量(图38A)或两个剂量(图38B)的BV2373和25μg或50μg的MATRIX-M^TM免疫食蟹猴后21天和35天的抗CoV S多肽IgG滴度。

图38C-图38D显示了用一个剂量(图38C)或两个剂量(图38D)的BV2373(5μg)和MATRIX-M^TM(25μg或50μg)免疫食蟹猴后21天和35天食蟹猴的hACE2抑制滴度。

图38E显示了在施用BV2373和MATRIX-M^TM后在食蟹猴中抗CoV S多肽IgG滴度与hACE2抑制滴度之间的显著相关性。显示了表4的第2-6组的数据。

图39显示了在用两个剂量的BV2373和MATRIX-M^TM免疫后或在用表4的恢复期人血清(第2、4和6组)免疫后35天食蟹猴的抗CoV S多肽滴度和hACE2抑制滴度。这些数据表明，用BV2373和MATRIX-M^TM免疫的食蟹猴的抗CoV S多肽和hACE2抑制滴度优于用恢复期血清免疫的食蟹猴。

图40A-图40B显示了用BV2373和MATRIX-M^TM免疫的食蟹猴的SARS-CoV-2中和滴度，如通过致细胞病变效应(CPE)(图40A)和噬斑减少中和测试(PRNT)(图40B)所测定的。

图41显示了评价包含BV2373和任选的MATRIX-M^TM的疫苗的安全性和功效的临床试验的施用时间。AESI表示特别关注的不良事件。MAEE表示医疗护理不良事件，并且SAE表示严重不良事件。

图42A-图42B显示了在评价包含BV2373和MATRIX-M^TM的疫苗的临床试验中患者经历的局部(图42A)和全身性(图42B)不良事件。在表5中标识了组A-E。数据表明，疫苗具有良好的耐受性和安全性。

图43A-图43B显示了在评价包含BV2373和MATRIX-M^TM的疫苗的临床试验中免疫参与者后21天和35天的抗CoV S多肽IgG(图43A)和中和(图43B)滴度。水平条分别表示四分位距(IRQ)和中值曲线下面积。须端点等于低于或高于中值±1.5倍IQR的最大值和最小值。恢复期血清图包括来自美国贝勒医学院(Baylor College of Medicine)的PCR确认的COVID-19参与者的样本(29个样本用于ELISA且32个样本用于微量中和(MN IC_>99))。COVID-19的严重性表示为住院治疗患者的红色标记(包括重症监护环境)、门诊治疗患者的蓝色标记(在急诊科收集的样品)和无症状(暴露)患者的绿色标记(从接触/暴露评估中收集的样品)。

图44A-图44C显示了在施用恢复期血清(图44A)、两个25μg剂量的BV2373(图44B)和两个剂量(5μg和25μg)的BV2373加MATRIX-M^TM(图44C)的患者中抗CoV S多肽IgG与中和抗体滴度之间的相关性。在用恢复期血清或佐剂化BV2373治疗的患者中在中和抗体滴度与抗CoV-S IgG滴度之间观察到很强的相关性，而在不存在佐剂的情况下用BV2373治疗的患者中并未观察到。

图45A-图45D显示了在A组(安慰剂，图45A)、B组(25μg BV2373，图45B)、C组(5μgBV2373和50μg MATRIX-M^TM，图45C)和D组(25μg BV2373和50μg MATRIX-M^TM，图45D)中在用BV2373刺激后参与者的产生T辅助1(Th1)细胞因子干扰素-γ(IFN-γ)、肿瘤坏死因子-α(TNF-α)以及白介素(IL)-2和T辅助2(Th2)细胞因子IL-5和IL-13指示的细胞因子的抗原特异性CD4+T细胞的频率。Th1细胞因子图中的“任何2”意指可以同时产生两种类型的Th1细胞因子的CD4⁺T细胞。“全部3”指示同时产生IFN-γ、TNF-α和IL-2的CD4⁺T细胞。Th2图中的“两者”意指可以同时产生Th2细胞因子IL-5和IL-13的CD4⁺T细胞。

图46A显示了含有信号肽的野生型SARS-CoV-2S多肽的一级结构，其关于SEQ IDNO:1进行编号。图46B显示了没有信号肽的野生型SARS-CoV-2S多肽的一级结构，其关于SEQID NO:2进行编号。

图47显示了3期临床试验中受试者的随机化，所述临床试验评价了与级分A和级分C iscom基质(MATRIX-M^TM)佐剂组合的BV2373的功效、免疫原性和安全性。

图48是Kaplan-Meyer图，其显示了受试者在接种与级分A和级分C iscom基质(MATRIX-M^TM)组合的BV2373或安慰剂后经历的症状性COVID-19的发生率(累积事件率(％))。

图49显示了与级分A和级分C iscom基质(MATRIX-M^TM)组合的BV2373针对包含具有SEQ ID NO:1的氨基酸序列的CoV S多肽的SARS-CoV-2或者包含具有氨基酸69、70和144的缺失以及N501Y、A570D、D614G、P681H、T716I、S982A和D1118H的突变的CoV S多肽的异源B.1.1.7SARS-CoV-2毒株的疫苗功效。

图50是显示了在使用与级分A和级分C iscom基质(MATRIX-M^TM)组合的BV2373(标记为“A”)或安慰剂(标记为“B”)的第一疫苗接种剂量(标记为“疫苗接种1”)和第二疫苗接种剂量(标记为“疫苗接种2”)之后受试者所经历的不良事件的图。

图51显示了BV2438 CoV S多肽的示意图。结构组件包括可切割信号肽(SP)、N末端结构域(NTD)、受体结合结构域(RBD)、亚结构域1和2(SD1和SD2)、S2切割位点(S2’)、融合肽(FP)、七肽重复区1(HR1)、中央螺旋(CH)、七肽重复区2(HR2)、跨膜结构域(TM)和胞质尾(CT)。与具有SEQ ID NO:1的氨基酸序列的CoV S多肽相比的氨基酸变化在线性图下方以黑色文本显示。

图52A显示了用考马斯蓝染色纯化的全长BV2438的还原SDS-PAGE凝胶，其显示预期分子量约170kD的主要蛋白质产物。图52B显示了扫描光密度测定法的图。图52C显示了BV2438的负染色透射电子显微照片。BV2438形成长度为15nm且宽度为11nm的界限清楚的灯泡形颗粒(左图)。三聚体展现出与PS-80胶束连接的8nm柔性接头(左图)。类平均图像显示rS-B.1.351三聚体与在2D图像上叠加的融合前SARS-CoV-2三聚体刺突蛋白胞外域(PDB ID6VXX)的低温EM解析结构的良好拟合(中间图)。右图显示了两个锚定至PS-80胶束中的BV2438三聚体。

图53显示了实例10的小鼠研究设计。在初次免疫/加强免疫方案中的第0天和第14天，使用重组S(rS)BV2438(SA)或BV2373(WU)蛋白的各种组合对小鼠组(n＝20只/组)进行免疫。小鼠用BV2438进行初次免疫和加强免疫，用BV2373进行初次免疫和加强免疫，用BV2373进行初次免疫并且用BV2438进行加强免疫，或用二价BV2373+BV2438进行初次免疫和加强免疫。对于每次单价免疫的抗原剂量为1μg rS，或对于二价免疫后的每个构建体为1μg rS(总共2μg rS)。所有抗原剂量均与5μg皂苷佐剂一起施用。对照组接受制剂缓冲液(安慰剂)。在图中列出的时间点收集血清和组织。

图54A-图54B显示了在实例10的小鼠研究的第21天收集的血清中的抗SARS-CoV-2S IgG血清滴度。ELISA用于测量针对Wuhan-Hu-1刺突蛋白(图54A)或B.1.351刺突蛋白(图54B)的抗体滴度。条形指示几何平均滴度(GMT)，并且误差条表示每组的95％置信区间(CI)。单独动物滴度用有色符号指示。图54C-图54D显示了在第21天收集的血清中的能够破坏SARS-CoV-2受体hACE2与Wuhan-Hu-1刺突蛋白(图54C)或B.1.351刺突蛋白(图54D)之间的结合的功能性抗体滴度(通过ELISA测量)。条形指示几何平均滴度(GMT)，并且误差条表示每组的95％置信区间(CI)。单独动物滴度用有色符号指示。图54E显示了使用PRNT测定来测定的在第32天从n＝5只动物/组收集的血清中的SARS-CoV-2中和抗体滴度。评价了血清中和SARS-CoV-2USA-WA1、B.1.351变体或B.1.1.7变体的能力。条形指示几何平均滴度(GMT)，并且误差条表示每组的95％置信区间(CI)。单独动物滴度用符号指示。通过对log₁₀转化的数据进行单因素方差分析与Tukey事后检验来计算统计显著性。

图55A-图55F显示了用基于Wuhan-Hu-1或B.1.351的SARS-CoV-2rS免疫针对用活的SARS-CoV-2B.1.351或B.1.1.7病毒攻击的保护性功效。研究设计描述于图53中。用活的SARS-CoV-2B.1.351(左图)或B.1.1.7(右图)攻击免疫的小鼠(n＝10只/组)。在攻击之后四天内每天对小鼠称重，并计算它们相对于攻击日的体重的体重减轻百分比。图55A和图55B用符号显示了平均体重减轻百分比。误差条表示平均值的标准误差。攻击后2天处死一半小鼠，并对肺组织进行噬斑形成测定以测定肺病毒滴度(图55C、图55D)。在攻击后4天处死其余小鼠。图55E和图55F显示了肺组织中SARS-CoV-2亚基因组RNA的水平，并表示为攻击后第2天RNA相对于相应安慰剂组中平均值的倍数变化。水平条表示每个时间点n＝5只小鼠的组平均倍数变化，并且误差条表示标准偏差。

图56A-图56H显示在小鼠中用BV2373或BV2438方案免疫后诱导的细胞介导的免疫。图56A显示了小鼠研究设计。在初次免疫/加强免疫方案中第0天和第21天，用基于BV2373或BV2438的SARS-CoV-2rS的各种组合对小鼠组(n＝8只/组)进行免疫。小鼠用BV2438进行初次免疫和加强免疫，用BV2373进行初次免疫和加强免疫，用BV2373进行初次免疫并且用BV2438进行加强免疫，或用二价BV2373和BV2438进行初次免疫和加强免疫。对于每次单价免疫的抗原剂量为1μg rS，或对于二价免疫后的每个构建体为1μg rS(总共2μgrS)。所有免疫均与5μg Matrix-M1佐剂一起施用。对照组接受制剂缓冲液(安慰剂，n＝5)。在第28天收获脾脏以用于细胞收集。用BV2373或BV2438刺激脾细胞，然后进行ELISA以确定IFN-γ阳性细胞为代表性Th1细胞因子(图56B)且IL-5阳性细胞为代表性Th2细胞因子(图56C)。来自图56B和图56C的数据用于计算对免疫的反应的Th1/Th2平衡(图56D)。图56E显示量化了使用细胞内细胞因子染色时对三种Th1细胞因子(IFN-γ、IL-2和TNF-α)染色呈阳性的多功能CD4+T细胞的数量并将其表示为每10⁶个CD4+T细胞中三重细胞因子阳性的细胞的数量。图56F显示了滤泡辅助性T细胞的量化。通过测定所有CD4+T细胞中PD-1+CXCR5+细胞的百分比来量化滤泡辅助性T细胞。图56G显示了通过使用流式细胞术确定CD19+B细胞中GL7+CD95+细胞的百分比来评价生发中心形成。灰色条表示平均值并且误差条表示标准偏差。单独动物数据用有色符号显示。门控策略的例子示于图56H。实验组之间的差异通过单因素方差分析与Tukey事后检验进行评价(图56B中的数据在分析前经过log₁₀转化)。P值<0.05被认为是统计上显著的；****＝p<0.0001。

图57A-图57E显示了来自用BV2373或BV2438免疫的CD4+和CD8+T细胞反应。在初次免疫/加强免疫方案中第0天和第21天，用BV2373或BV2438的各种组合对小鼠组(n＝8只/组)进行免疫。小鼠用BV2438进行初次免疫和加强免疫，用BV2373进行初次免疫和加强免疫，用BV2373进行初次免疫并且用BV2438进行加强免疫，或用二价BV2373和BV2438进行初次免疫和加强免疫。对于每次单价免疫的抗原剂量为1μg rS，或对于二价免疫后的每个构建体为1μg rS(总共2μg rS)。所有抗原剂量均与5μg皂苷佐剂一起施用。对照组接受制剂缓冲液(安慰剂，n＝5)。在第28天收获脾脏以用于细胞收集。用rS-WU1或rS-B.1.351刺激分离的脾细胞，然后进行细胞内细胞因子染色以确定CD4+T细胞是否对IFN-γ(图57A)、IL-2(图57B)、TNF-α(图57C)或IL-4(图57D)呈阳性。为了检查CD8+T细胞反应，用对应于整个Wuhan-Hu-1刺突蛋白序列的肽池刺激细胞，然后对IFN-γ、IL-2和TNF-α进行ICS(图57E)。

图58A-图58G显示了在狒狒中用BV2373免疫后大约一年一个或两个加强BV2438剂量的免疫原性。图58A显示了研究设计。小的狒狒群组(n＝2-3只/组)在第0天和第21天(分别为第0周和第3周)用1μg、5μg或25μg BV2373与皂苷佐剂一起或25μg未佐剂化BV2373进行初始免疫。大约1年后，所有动物在第318天和第339天(分别为第45周和第48周)用一个或两个剂量的3μg BV2438与50μg皂苷佐剂一起进行加强免疫。图58B显示了研究过程中的抗CoVS IgG滴度。单独动物的滴度随时间显示，不同颜色的符号和线条表示初始rS-WU1免疫系列的不同剂量组。分析BV2438加强免疫前(第303天)以及加强免疫后7、21、35和81天收集的血清以通过ELISA测定抗rS-WU1(图58C)和抗rS-B.1.351(图58D)IgG滴度(水平线表示平均值)，通过ELISA测定能够破坏rS-WU1或rS-B.1.351与hACE2受体之间相互作用的抗体滴度(图58E，水平线表示平均值)，以及使用PRNT测定来测定能够中和SARS-CoV-2毒株USA-WA1、B.1.351和B.1.1.7的抗体滴度(图58F，灰色条表示几何平均值，并且误差条表示95％置信区间)。用BV2373或BV2438刺激后，用细胞内细胞因子染色评价对3种Th1细胞因子(IFN-γ、IL-2和TNF-α)呈阳性的多功能CD4+T细胞的存在(图58G)。灰色条表示平均值，并且有色符号表示单独动物数据。

图59A-图59G显示了狒狒中对BV2438加强免疫的单独细胞因子反应。小的狒狒群组(n＝2-3只/组)在第0天和第21天(分别为第0周和第3周)用1μg、5μg或25μg BV2373与50μg皂苷佐剂一起或25μg未佐剂化BV2373进行免疫。大约1年后，所有动物在第318天和第339天(分别为第45周和第48周)用一个或两个剂量的3μg BV2438与50μg皂苷佐剂一起进行加强免疫。在加强免疫前(第303天；第43周)、第一次rS-B.1.351加强免疫后7天(第325天；第46周)和第一次rS-B.1.351加强免疫后35天(第353天；第50周)收集PBMC。用BV2373或BV2438刺激PBMC并对其进行ELISA以测量作为Th1细胞因子的IFN-γ产生性细胞(图59A)和作为Th2细胞因子的IL-4产生性细胞(图59B)。也用BV2373或BV2438刺激CD4+T细胞，然后对其进行ICS以测量产生IFN-γ(图59C)、IL-2(图59D)、TNF-α(图59E)、IL-5(图59F)和IL-13(图59G)的细胞。

图60A-图60B显示了来自用BV2373免疫的人受试者的SARS-CoV-2变体中和滴度。与B.1.1.7(图60A)和B.1.351(图60B)相比，对来自临床研究参与者(n＝30)的血清样品进行PRNT测定以确定针对USA-WA1的中和抗体的存在。单独受试者的滴度用黑色圆圈显示，线条将个体针对USA-WA1的滴度与他们针对相应变体的滴度连接在一起。

图61A-图61B显示了对于以下SARS-CoV-2变体在用BV2373和皂苷佐剂加强免疫之前和之后的抗S蛋白IgG滴度：(i)与具有SEQ ID NO:1的氨基酸序列的蛋白质相比，具有含有D614G突变的CoV S多肽的SARS-CoV-2病毒；(ii)SARS-CoV-2α毒株、SARS-CoV-2β毒株和SARS-CoV-2δ毒株。图61A显示了从第35天到第217天的倍数增加。图61B显示了从第189天到第217天的倍数增加。

图62A-图62B显示了对于以下SARS-CoV-2变体在用BV2373和皂苷佐剂加强免疫之前和之后的功能性hACE2抑制：(i)与具有SEQ ID NO:1的氨基酸序列的蛋白质相比，具有含有D614G突变的CoV S多肽的SARS-CoV-2病毒；(ii)SARS-CoV-2α毒株、SARS-CoV-2β毒株和SARS-CoV-2δ毒株。图62A显示了从第35天到第217天的倍数增加。图62B显示了从第189天到第217天的倍数增加。

图63显示了实例11中描述的试验参与者的加强免疫给药的图。

图64A-图64B显示了实例11中所述试验的B2组中患者的局部(图64A)和全身性(图64B)反应原性。

图65显示了在实例11中描述的患者截至研究日对原始SARS-CoV-2毒株的血清IgG滴度。

图66显示了在实例11中描述的患者截至研究日对于原始SARS-CoV-2毒株的中和抗体活性。

图67显示了实例11的包含BV2373和皂苷佐剂的免疫原性组合物针对抗含有D614G突变的SARS-CoV-2毒株和B.1.617.2(δ变体)的中和抗体99(neut99)值。

发明详述

定义

如本申请和所附权利要求所用，单数形式“一个/一种(a、an)”和“所述(the)”包括复数指代物，除非上下文另外明确地规定。因此，例如，提及“一种蛋白质”可以指一种蛋白质或这种蛋白质的混合物，并且提及“所述方法”包括提及本领域技术人员已知的等同步骤和/或方法，等等。

如本申请所用，术语“佐剂”是指当与免疫原组合使用时增加或以其他方式改变或修饰针对所述免疫原诱导的免疫反应的化合物。免疫反应的修饰可以包括强化或扩大抗体免疫反应和细胞免疫反应中任一者或两者的特异性。

如本申请所用，当在数值之前时，术语“约”或“大约”表示所述值加或减10％的范围。例如，“约100”包括90和110。

如本申请所用，术语“免疫原”、“抗原”和“表位”是指能够引发免疫反应的物质，例如蛋白质(包括糖蛋白)和肽。

如本申请所用，“免疫原性组合物”是包含抗原的组合物，其中向受试者施用所述组合物导致在所述受试者中产生针对所述抗原的体液免疫反应和/或细胞免疫反应。

如本申请所用，“亚基”组合物(例如疫苗)包括一种或多种来自病原体的选定抗原，但不是全部抗原。这样的组合物基本上不含完整的病毒或此类细胞或颗粒的裂解物，并且通常由至少部分纯化的(通常基本上纯化的)来自所述病原体的免疫原性多肽制备。通常使用杆状病毒系统，通常以重组方式制备本申请所公开的亚基组合物中的抗原。

如本申请所用，“基本上”是指这样分离物质(例如化合物、多核苷酸或多肽)，使得所述物质形成包含其的样品的大多数百分比。例如，在样品中，基本上纯化的组分包含样品的85％，优选85％-90％，更优选至少95％-99.5％，以及最优选至少99％。如果组分基本上被替代，则在样品中剩余的量为小于或等于约0.5％至约10％，优选小于约0.5％至约1.0％。

如本申请所用，术语“治疗”(“treat”、“treatment”和“treating”)是指用于获得有益或希望的结果(例如，临床结果)的方法。出于本申请公开文本的目的，有益或希望的结果可以包括抑制或压制感染或疾病的开始或进展；改善感染或疾病的症状或减轻其发展；或其组合。

如本申请所用，“预防”(“prevention”)与“防预”(“prophylaxis”)可互换使用，并且可以意指完全预防感染或疾病，或预防这种感染或疾病的症状的发展；延迟感染或疾病或其症状的发作；或降低随后发展的感染或疾病或其症状的严重性。

如本申请所用，“有效剂量”或“有效量”是指免疫原的这样的量，所述量足以诱导减轻病原体感染的至少一种症状的免疫反应。可以例如通过测量中和分泌抗体和/或血清抗体的量，例如通过噬斑中和、补体结合、酶联免疫吸附(ELISA)或微量中和测定来确定有效剂量或有效量。

如本申请所用，术语“疫苗”是指免疫原性组合物，如源自病原体的免疫原，其用于诱导针对病原体的免疫反应。免疫反应可以包括抗体和/或细胞介导的反应的形成。取决于上下文，术语“疫苗”也可以是指施用至受试者以产生免疫反应的免疫原悬浮液或溶液。优选地，疫苗诱导有效预防SARS-CoV-2或其变体感染的免疫反应。

如本申请所用，术语“受试者”包括人和其他动物。通常，受试者是人。例如，受试者可以是成人、青少年、儿童(2岁至14岁)、婴儿(出生至2岁)或新生儿(最大2个月)。在特定方面，受试者最大4个月大或最大6个月大。在各个方面，成人是约65岁以上、或约60岁以上的年长者。在各个方面，受试者是孕妇或打算怀孕的妇女。在其他方面，受试者不是人；例如为非人灵长类动物；例如，狒狒、黑猩猩、大猩猩或猕猴。在某些方面，受试者可以是宠物，诸如狗或猫。

在各个方面，受试者是免疫功能受损的。在实施方案中，向免疫功能受损的受试者施用引起免疫抑制的药物。引起免疫抑制的药物的非限制性例子包括皮质类固醇(例如，泼尼松)、烷基化剂(例如，环磷酰胺)、抗代谢物(例如，硫唑嘌呤或6-巯基嘌呤)、移植相关免疫抑制药物(例如，环孢菌素、他克莫司、西罗莫司或吗替麦考酚酯)、米托蒽醌、化疗剂、甲氨蝶呤、肿瘤坏死因子(TNF)阻断剂(例如，依那西普、阿达木单抗、英夫利昔单抗)。在实施方案中，免疫功能受损的受试者感染了病毒(例如，人类免疫缺陷病毒或爱泼斯坦-巴尔病毒)。在实施方案中，所述病毒是呼吸道病毒，如呼吸道合胞病毒、流感病毒、副流感病毒、腺病毒或小核糖核酸病毒。在实施方案中，免疫功能受损的受试者患有获得性免疫缺陷综合征(AIDS)。在实施方案中，免疫功能受损的受试者是患有人类免疫缺陷病毒(HIV)的人。在实施方案中，免疫功能受损的受试者是由于被设计用于预防炎症或防止移植排斥的治疗方案而免疫功能受损。在实施方案中，免疫功能受损的受试者是已接受移植的受试者。在实施方案中，免疫功能受损的受试者已进行放射疗法或脾切除术。在实施方案中，免疫功能受损的受试者已被诊断为患有癌症、自身免疫性疾病、肺结核、物质使用障碍(例如，酒精、阿片类药物或可卡因使用障碍)、中风或脑血管疾病、实体器官或血液干细胞移植、镰状细胞病、地中海贫血、自身免疫性淋巴组织增生综合征(ALPS)、自身免疫性多腺体综合征1型(APS-1)、B细胞扩增伴NF-κB和T细胞无能(BENTA)疾病、胱天蛋白酶-8缺乏状态(CEDS)、慢性肉芽肿病(CGD)、常见变异型免疫缺陷(CVID)、先天性中性粒细胞减少综合征、细胞毒性T淋巴细胞相关抗原4(CTLA-4)缺陷、DOCK8缺陷、GATA2缺陷、糖基化障碍伴免疫缺陷、高免疫球蛋白E综合征(HIES)、高免疫球蛋白M综合征、糖尿病、1型糖尿病、2型糖尿病、干扰素γ缺陷、白介素12缺陷、白介素23缺陷、白细胞粘附缺陷、脂多糖反应性米色锚样蛋白(LRBA)缺陷、PI3激酶疾病、PLCG2相关抗体缺陷和免疫失调(PLAID)、严重综合性免疫缺陷(SCID)、STAT3显性负性疾病、STAT3功能获得疾病、疣、低丙种球蛋白血症、感染和无效生成性慢性粒细胞缺乏(WHIM)综合征、Wisckott-Aldrich综合征(WAS)、X连锁无丙种球蛋白血症(XLA)、X连锁淋巴组织增生性疾病(XLP)、尿毒症、营养不良或XMEN疾病。在实施方案中，免疫功能受损的受试者是当前吸烟者或曾吸烟者。在实施方案中，免疫功能受损的受试者患有B细胞缺陷、T细胞缺陷、巨噬细胞缺陷、细胞因子缺陷、吞噬细胞缺陷、吞噬细胞功能障碍、补体缺陷或其组合。

在实施方案中，受试者超重或肥胖。在实施方案中，超重受试者的体重指数(BMI)≥25kg/m²且<30kg/m²。在实施方案中，肥胖受试者的BMI≥30kg/m²。在实施方案中，受试者患有精神健康病症。在实施方案中，精神健康病症是抑郁症、精神分裂症或焦虑。

如本申请所用，术语“医药上可接受的”意指由美国联邦政府或州政府的监管机构批准或在美国药典、欧洲药典或其他公认药典中列出用于哺乳动物，更特别地用于人。这些组合物可以用作用于在脊椎动物中诱导保护性免疫反应的疫苗和/或抗原组合物。

如本申请所用，术语“约”意指所指数值加或减10％。

如本申请所用，术语“NVX-CoV2373”是指包含BV2373刺突糖蛋白(SEQ ID NO:87)以及级分A和级分C iscom基质(例如，MATRIX-M^TM)的疫苗组合物。

如本申请所用，术语“修饰”当提及CoV S多肽时是指CoV S多肽的一个或多个氨基酸的突变、缺失或添加。CoV S多肽内修饰的位置可以基于将多肽的序列与SEQ ID NO:1(含有信号肽的CoV S多肽)或SEQ ID NO:2(缺少信号肽的成熟CoV S多肽)比对来确定。

在本申请中与“异源SARS-CoV-2毒株”可互换使用的术语SARS-CoV-2的变体是包含CoV S多肽的SARS-CoV-2病毒，与具有SEQ ID NO:1或SEQ ID NO:2的氨基酸序列的CoV S多肽相比，所述多肽含有至少约2、至少约3、至少约4、至少约5、至少约6、至少约7、至少约8、至少约9、至少约10、至少约11、至少约12、至少约13、至少约14、至少约15、至少约16、至少约17、至少约18、至少约19、至少约20、至少约21、至少约22、至少约23、至少约24、至少约25、至少约26、至少约27、至少约28、至少约29、至少约30、至少约31、至少约32、至少约33、至少约34或至少约35个修饰、在约2与约35个之间的修饰、在约5与约10个之间的修饰、在约5与约20个之间的修饰、在约10与约20个之间的修饰、在约15与约25个之间的修饰、在约20与30个之间的修饰、在约20与约40个之间的修饰、在约25与约45个之间的修饰。在实施方案中，所述异源SARS-CoV-2毒株是包含CoV S多肽的SARS-CoV-2病毒，所述多肽与具有SEQ ID NO:1或SEQ ID NO:2的氨基酸序列的CoV S多肽具有至少约70％、至少约75％、至少约80％、至少约85％、至少约90％、至少约95％、至少约96％、至少约97％、至少约98％或至少约99％同一性。在实施方案中，所述异源SARS-CoV-2毒株是包含CoV S多肽的SARS-CoV-2病毒，所述多肽与具有SEQ ID NO:1或SEQ ID NO:2的氨基酸序列的CoV S多肽具有在约70％与约99.9％之间的同一性。在实施方案中，所述异源SARS-CoV-2毒株是包含CoV S多肽的SARS-CoV-2病毒，所述多肽与具有SEQ ID NO:1或SEQ ID NO:2的氨基酸序列的CoV S多肽具有在约70％与约99.5％之间的同一性。在实施方案中，所述异源SARS-CoV-2毒株是包含CoV S多肽的SARS-CoV-2病毒，所述多肽与具有SEQ ID NO:1或SEQ ID NO:2的氨基酸序列的CoV S多肽具有在约90％与约99.9％之间的同一性。在实施方案中，所述异源SARS-CoV-2毒株是包含CoV S多肽的SARS-CoV-2病毒，所述多肽与具有SEQ ID NO:1或SEQ ID NO:2的氨基酸序列的CoV S多肽具有在约90％与约99.8％之间的同一性。在实施方案中，所述异源SARS-CoV-2毒株是包含CoV S多肽的SARS-CoV-2病毒，所述多肽与具有SEQ ID NO:1或SEQ ID NO:2的氨基酸序列的CoV S多肽具有在约95％与约99.9％之间的同一性。在实施方案中，所述异源SARS-CoV-2毒株是包含CoV S多肽的SARS-CoV-2病毒，所述多肽与具有SEQ ID NO:1或SEQID NO:2的氨基酸序列的CoV S多肽具有在约95％与约99.8％之间的同一性。在实施方案中，所述异源SARS-CoV-2毒株是包含CoV S多肽的SARS-CoV-2病毒，所述多肽与具有SEQ IDNO:1或SEQ ID NO:2的氨基酸序列的CoV S多肽具有在约95％与约99％之间的同一性。

术语“B.1.1.7SARS-CoV-2毒株”(也称为“α”毒株)是指具有包含氨基酸69、70和144的缺失以及N501Y、A570D、D614G、P681H或P681R、T716I、S982A和D1118H的突变的CoV S多肽的异源SARS-CoV-2毒株，其中所述CoV S多肽关于具有SEQ ID NO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。B.1.1.7SARS-CoV-2毒株的CoV S多肽可以任选地含有氨基酸145的缺失、E484K、L432R或S494P的突变或其组合。

术语“B.1.351SARS-CoV-2毒株”(也称为“β”毒株)是指具有包含D80A、K417N、E484K、N501Y、D614G和A701V的突变的CoV S多肽的异源SARS-CoV-2毒株，其中所述CoV S多肽关于具有SEQ ID NO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。B.1.617.2SARS-CoV-2毒株的CoV S多肽可以任选地含有以下突变中的一个或多个：D215G；L242H；R246I；或241-243的1、2或3个氨基酸的缺失，其中所述CoV S多肽关于具有SEQ IDNO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。在实施方案中，所述β毒株的CoV S多肽包含D80A、D215G、L242H、K417N、E484K、N501Y、D614G和A701V的突变，其中所述CoV S多肽关于具有SEQ ID NO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。在实施方案中，所述β毒株的CoV S多肽包含D80A、D215G、氨基酸241-243的1、2或3个氨基酸的缺失、K417N、E484K、N501Y、D614G和A701V的突变，其中所述CoV S多肽关于具有SEQ ID NO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。在实施方案中，所述β毒株包含D80A、L242H、R246I、N501Y、K417N、E484K、D614G和A701V的突变，其中所述CoV S多肽关于具有SEQ IDNO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。

术语“P.1SARS-CoV-2毒株”(也称为“γ”毒株)是指具有含有突变L18F、T20N、P26S、D138Y、R190S、K417T、E484K、N501Y、D614G、H655Y、T1027I和V1176F的CoV S多肽的异源SARS-CoV-2毒株，其中所述CoV S多肽关于具有SEQ ID NO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。

术语“Cal.20C SARS-CoV-2毒株”是指具有含有突变S13I、W152C和L452R的CoV S多肽的异源SARS-CoV-2毒株，其中所述CoV S多肽关于具有SEQ ID NO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。

术语“B.1.617.2毒株”(也称为“δ”毒株)是指具有包含氨基酸157和158的缺失以及T19R、E156G、L452R、T478K、D614G、P681R和D950N的突变的CoV S多肽的异源SARS-CoV-2毒株，其中所述CoV S多肽关于具有SEQ ID NO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。B.1.617.2SARS-CoV-2毒株的CoV S多肽可以任选地含有以下突变中的一个或多个：G142D；W64H；H66W；V70F；T95I；Y145H；D213V；L214R；A222V；W258I或W258L；K417N；N439K；E484K或E484Q；N501Y；和Q613H，其中所述CoV S多肽关于具有SEQ ID NO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。在实施方案中，所述δ毒株包含含有氨基酸157和158的缺失以及T19R、G142D、E156G、L452R、T478K、D614G、P681R和D950N的突变的CoV S多肽。在实施方案中，所述δ毒株包含含有氨基酸157和158的缺失以及T19R、T95I、G142D、Y145H、E156G、A222V、K417N L452R、T478K、D614G、P681R和D950N的突变的CoV S多肽，其中所述CoV S多肽关于具有SEQ ID NO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。在实施方案中，所述δ毒株包含含有氨基酸157和158的缺失以及T19R、G142D、E156G、W258I、K417N、N439K、L452R、T478K、E484K、N501Y、D614G、P681R和D950N的突变的CoV S多肽，其中所述CoV S多肽关于具有SEQ ID NO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。在实施方案中，所述δ毒株包含含有氨基酸157和158的缺失以及T19R、W64H、H66W、G142D、E156G、D213V、L214R、W258I、K417N、N439K、L452R、T478K、E484K、N501Y、D614G、P681R和D950N的突变的CoV S多肽，其中所述CoV S多肽关于具有SEQ ID NO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。在实施方案中，所述δ毒株包含含有氨基酸157和158的缺失以及T19R、G142D、E156G、K417N、L452R、T478K、E484Q、D614G、P681R和D950N的突变的CoV S多肽，其中所述CoV S多肽关于具有SEQ ID NO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。

术语“B.1.525毒株”(也称为“η”毒株)是指具有含有突变Q52R；A67V；E484K；D614G；Q677H；F888L；以及氨基酸69、70、144、145的1、2、3或4个的缺失的CoV S多肽的异源SARS-CoV-2毒株，其中所述CoV S多肽关于具有SEQ ID NO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。

术语“B.1.526毒株”(也称为“ι”毒株)是指具有含有突变L5F；T95I；D253G；E484K；D614G；和A701V的CoV S多肽的异源SARS-CoV-2毒株，其中所述CoV S多肽关于具有SEQ IDNO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。

术语“B.1.617.1毒株”(也称为“κ”毒株)是指具有含有突变L452R；E484Q；D614G；P681R；和Q1071H的CoV S多肽的异源SARS-CoV-2毒株，其中所述CoV S多肽关于具有SEQ IDNO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。

术语“C.37毒株”(也称为“λ”毒株)是指具有含有突变G75V；T76I；R246N；L452Q；F490S；D614G；T859N；以及氨基酸247-253的1、2、3、4、5或6个的缺失的CoV S多肽的异源SARS-CoV-2毒株，其中所述CoV S多肽关于具有SEQ ID NO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。

术语“B.1.621毒株”(也称为“μ”毒株)是指具有含有突变T95I；Y144S；Y145N；R346K；E484K；N501Y；D614G；P681H；和D950N的CoV S多肽的异源SARS-CoV-2毒株，其中所述CoV S多肽关于具有SEQ ID NO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。

本申请所述的术语免疫原性组合物或疫苗组合物的“功效”是指与未施用所述免疫原性组合物的组相比，施用了免疫原性组合物的组中疾病(例如，COVID-19)的百分比降低。在实施方案中，功效(E)使用以下方程式计算：E(％)＝(1-RR)×100，其中RR＝施用了所述免疫原性组合物的组与未施用所述免疫原性组合物的组之间的发病率的相对风险。在实施方案中，本申请所述的免疫原性组合物对SARS-CoV-2病毒或异源SARS-CoV-2毒株的功效为至少约50％、至少约55％、至少约60％、至少约65％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％、至少约91％、至少约92％、至少约93％、至少约94％、至少约95％、至少约96％、至少约97％、至少约98％、至少约99％、在约50％与约99％之间、在约50％与约98％之间、在约60％与约99％之间、在约60％与约98％之间、在约70％与约98％之间、在约70％与约95％之间、在约70％与约99％之间、在约80％与约99％之间、在约80％与约98％之间、在约80％与约95％之间、在约85％与约99％之间、在约85％与约98％之间、在约85％与约95％之间、在约90％与约95％之间、在约90％与98％之间或在约90％与约99％之间。

对SARS-CoV-2或其变体呈“阳性”的受试者具有对于SARS-CoV-2或其变体的阳性PCR或血清学测试。阳性PCR测试检测到来自SARS-CoV-2或其变体的遗传物质。阳性血清学测试显示出存在针对SARS-CoV-2蛋白、通常来自SARS-CoV-2或其变体的核衣壳蛋白的抗体。

术语“无症状”是指对SARS-CoV-2或其SARS-CoV-2变体呈阳性但未经历任何COVID-19的症状的受试者。

术语“轻度”在指COVID-19时是指具有对于SARS-CoV-2或其变体的阳性PCR或血清学测试并且具有以下症状中的一种或多种的受试者：(i)发热；(ii)新发作的咳嗽；(iii)或选自以下的两种另外的COVID-19症状：新发作或加重的呼吸急短促或呼吸困难；疲劳；全身肌肉或身体疼痛；头痛；失去味觉或嗅觉；喉咙痛、充血或流鼻涕；或恶心、呕吐或腹泻。

术语“中度”在指COVID-19时是指具有对于SARS-CoV-2或其变体的阳性PCR或血清学测试并且具有以下症状中的一种或多种的受试者：(i)≥38.4℃的高热持续三天或更长时间；(ii)任何明显下呼吸道感染(LRTI)的迹象，其中所述迹象选自：(a)用力或不用力时呼吸短促；(b)呼吸急促(休息时每分钟24至29次呼吸)；(c)SpO2为94％至95％；(d)与肺炎或LRTI一致的异常胸部X光或计算机断层扫描(CT)；或(e)肺部听诊时的附加音(例如，湿罗音/罗音、喘息、干罗音、胸膜摩擦音、喘鸣)。

术语“重度”在指COVID-19时是指具有对于SARS-CoV-2或其变体的阳性PCR或血清学测试以及以下症状中的一种或多种的受试者：(i)休息时呼吸急促≥30次呼吸/分；(ii)静息心率≥125次心跳/分；(iii)SpO2≤93％或PaO2/FiO2<300mmHg；(iv)需要高流量氧疗法或无创通气、无创正压通气(例如，持续气道正压通气(CPAP)或双水平气道正压通气(BiPAP))；(v)需要机械通气或体外膜肺氧合(ECMO)；(vi)选自以下的一种或多种主要器官系统功能障碍或衰竭：(a)急性呼吸衰竭，包括急性呼吸窘迫综合征(ARDS)；(b)急性肾衰竭；(c)急性肝衰竭；(d)急性右心或左心衰竭；(e)感染性或心原性休克(其中休克定义为收缩压(SBP)<90mmHg或舒张压(DBP)<60mmHg)；(f)急性中风(缺血性或出血性)；(g)急性血栓事件，如急性心肌梗死(AMI)、深静脉血栓形成(DVT)或肺栓塞(PE)；(h)需要血管加压药、全身性皮质类固醇或血液透析；(vii)入住重症监护室；或(viii)死亡。

含有冠状病毒(CoV)刺突(S)蛋白的疫苗组合物

本申请公开文本提供了非天然存在的冠状病毒(CoV)刺突(S)多肽、含有CoV S多肽的纳米颗粒以及含有非天然存在的CoV S多肽或含有CoV S多肽的纳米颗粒的免疫原性组合物和疫苗组合物。在实施方案中，本申请提供了使用CoV S多肽、纳米颗粒、免疫原性组合物和疫苗组合物来刺激免疫反应的方法。

本申请还提供了制造纳米颗粒和疫苗组合物的方法。有利地，所述方法提供了基本上没有被其他蛋白质污染的纳米颗粒，所述其他蛋白质是诸如与蛋白质在昆虫细胞中的重组表达相关的蛋白质。在实施方案中，表达发生在杆状病毒/Sf9系统中。

CoV S多肽抗原

本申请公开文本的疫苗组合物含有非天然存在的CoV S多肽。CoV S多肽可以源自冠状病毒，包括但不限于SARS-CoV-2，例如源自SARS-CoV-2、源自MERS CoV和源自SARSCoV。在实施方案中，所述CoV S多肽源自SARS-CoV-2的变体。在实施方案中，SARS-CoV-2的变体是SARS-CoV-2VUI 202012/01、B.1.1.7(也称为“501Y.V1”和“α”)、B.1.351(也称为“501Y.V2”和“β”)、B.1.617.2(也称为“δ”)、Cal.20C(也称为“ε”)或P.1(也称为“γ”)。SARS-CoV-2的变体由世界卫生组织(WHO)标记(例如，α、β、γ、δ等)、其以系统发育归类命名全球暴发(PANGO)谱系、其GISAID进化枝或其Nextstrain进化枝命名。

下表提供了SARS-CoV-2毒株的列表：

在实施方案中，所述SARS-CoV-2病毒具有含有SEQ ID NO:1的氨基酸序列的CoV S多肽，并且SARS-CoV-2的变体包含如下CoV S多肽，与SEQ ID NO:1相比，所述多肽具有至少约1、至少约2、至少约3、至少约4、至少约5、至少约6、至少约7、至少约8、至少约9、至少约10、至少约11、至少约12、至少约13、至少约14、至少约15、至少约16、至少约17、至少约18、至少约19、至少约20、至少约21、至少约22、至少约23、至少约24、至少约25、至少约26、至少约27、至少约28、至少约29、至少约30、至少约31、至少约32、至少约33、至少约34、至少约35个修饰、至少约36、至少约37、至少约38、至少约39、至少约40、至少约41、至少约42、至少约43、至少约44、至少约45、至少约46、至少约47、至少约48、至少约49、至少约50、至少约51、至少约52、至少约53、至少约54、至少约55、至少约56、至少约57、至少约58、至少约59或至少约60个修饰。

与SARS-CoV S相比，SARS-CoV-2S蛋白在S1/S2切割位点中具有四个氨基酸的插入，从而产生多碱性RRAR弗林蛋白酶样切割基序。SARS-CoV-2S蛋白被合成为无活性前体(S0)，其在弗林蛋白酶切割位点处被蛋白水解切割成S1和S2亚基，这些亚基保持非共价连接以形成融合前三聚体。SARS-CoV-2S蛋白的S2结构域包含融合肽(FP)、两个七肽重复区(HR1和HR2)、跨膜(TM)结构域和胞质尾(CT)。SARS-CoV-2S蛋白的S1结构域折叠成四个不同的结构域：N末端结构域(NTD)和含有受体结合结构域(RBD)的C末端结构域、以及两个亚结构域SD1和SD2。融合前SARS-CoV-2S蛋白三聚体在S蛋白受体结合和切割后经历从融合前到融合后构象的结构重排。

在实施方案中，由于翻译后糖基化，所述CoV S多肽是糖蛋白。所述糖蛋白包含信号肽、S1亚基、S2亚基、NTD、RBD、两个亚结构域(SD1和SD2，在图46A-图46B中标记为SD1/2并且在本申请中称为“SD1/2”)、完整或修饰的融合肽，HR1结构域、HR2结构域、TM和CD中的一种或多种。在实施方案中，每个结构域的氨基酸在图2和图46A(根据SEQ ID NO:1示出)、图46B(根据SEQ ID NO:2示出)和图3(对应于SEQ ID NO:1示出)中给出。在实施方案中，每个结构域可以与如在SEQ ID NO:1或SEQ ID NO:2中的每个结构域的序列具有至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、至少99％或至少99.5％同一性。与SEQ ID NO:1或SEQ ID NO:2中所示的那些相比，每个结构域可以具有多达约1、多达约2、多达约3、多达约4、多达约5、多达约10、多达约20、多达约30、多达约35、多达约40、多达约45、多达约50、多达约55、多达约60、多达约65或多达约70个氨基酸的缺失、插入或突变。与SEQ ID NO:1或SEQ ID NO:2中所示的那些相比，每个结构域可以具有在约1与约5个之间的氨基酸、在约3与约10个之间的氨基酸、在约5与约10个之间的氨基酸、在约8与约12个之间的氨基酸、在约10与约15个之间的氨基酸、在约12与约17个之间的氨基酸、在约15与约20个之间的氨基酸、在约18与约23个之间的氨基酸、在约20与约25个之间的氨基酸、在约22与约27个之间的氨基酸、在约25与约30个之间的氨基酸、在约30与约35个之间的氨基酸、在约35与约40个之间的氨基酸、在约40与约45个之间的氨基酸、在约45与约50个之间的氨基酸、在约50与约55个之间的氨基酸或在约55与约60个之间的氨基酸的缺失、插入或突变。注意，图2和图3展示了成熟肽中不存在的13个氨基酸的N末端信号肽。所述CoVS多肽可以用于刺激针对天然CoV刺突(S)多肽的免疫反应。

在实施方案中，修饰天然CoV刺突(S)多肽(SEQ ID NO:2)，从而产生非天然存在的CoV刺突(S)多肽(图1)。在实施方案中，所述CoV刺突(S)糖蛋白包含S1亚基和S2亚基，其中所述S1亚基包含NTD、RBD、SD1/2和无活性弗林蛋白酶切割位点(氨基酸669-672)，并且其中所述S2亚基包含氨基酸973和974的突变；

其中所述NTD(氨基酸1-318)任选地包含选自以下的一个或多个修饰：

(b)在氨基酸132之后1、2、3或4个氨基酸的插入；以及

其中所述S2亚基任选地包含选自以下的一个或多个修饰：

(c)从跨膜和胞质结构域(TMCT)(氨基酸1201-1260)中的一个或多个氨基酸的缺失，

其中所述CoV S糖蛋白的氨基酸关于SEQ ID NO:2进行编号。

图3显示了称为BV2378的CoV S多肽，其具有无活性弗林蛋白酶切割位点、缺失的融合肽(例如，氨基酸819-828的缺失)、K986P和V987突变，其中氨基酸关于SEQ ID NO:1进行编号。成熟BV2378多肽缺少信号肽的一个或多个氨基酸，所述信号肽为SEQ ID NO:1的氨基酸1-13。

在实施方案中，本申请所述的CoV S多肽以融合前构象存在。在实施方案中，本申请所述的CoV S多肽包含柔性HR2结构域。除非另外提及，否则结构域的柔性通过透射电子显微术(TEM)和2D类别取平均值来确定。电子密度的降低对应于柔性结构域。

CoV S多肽抗原-对S1亚基的修饰

在实施方案中，所述CoV S多肽含有对具有SEQ ID NO:121的氨基酸序列的S1亚基的一个或多个修饰。

所述S1亚基的氨基酸序列(SEQ ID NO:121)示于下文。

QCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRAR

SEQ ID NO:121的加下划线的区域表示S1亚基内的可被修饰的氨基酸。

在实施方案中，本申请所述的CoV S多肽包含与SEQ ID NO:1或SEQ ID NO:2的S1亚基具有至少95％、至少96％、至少97％、至少98％、至少99％或至少99.5％同一性的S1亚基。所述S1亚基可以具有与SEQ ID NO:1或SEQ ID NO:2的S1亚基的氨基酸序列相比多达约1、多达约2、多达约3、多达约4、多达约5、多达约10、多达约15、多达约20、多达约25或多达约30个氨基酸的缺失、插入或突变。所述S1亚基可以具有与SEQ ID NO:1或SEQ ID NO:2的S1亚基相比在约1与约5个之间的氨基酸、在约3与约10个之间的氨基酸、在约5与10个之间的氨基酸、在约8与12个之间的氨基酸、在约10与15个之间的氨基酸、在约12与17个之间的氨基酸、在约15与20个之间的氨基酸、在约18与23个之间的氨基酸、在约20与25个之间的氨基酸、在约22与约27个之间的氨基酸或在约25与30个之间的氨基酸的缺失、插入或突变。

在实施方案中，所述S1亚基可以含有表1A中所示的修饰的任何组合。

表1A

CoV S多肽抗原-对S1亚基-NTD的修饰

在实施方案中，所述CoV S多肽含有对NTD的一个或多个修饰。在实施方案中，所述NTD具有SEQ ID NO:118的氨基酸序列，其对应于SEQ ID NO:1的氨基酸14-305或SEQ IDNO:2的氨基酸1-292。

NTD的氨基酸序列(SEQ ID NO:118)示于下文。

QCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKS

在实施方案中，所述NTD具有SEQ ID NO:45的氨基酸序列，其对应于SEQ ID NO:1的氨基酸14至331或SEQ ID NO:2的氨基酸1-318。NTD的氨基酸序列(SEQ ID NO:45)示于下文。

QCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNV

TWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPN

在实施方案中，所述NTD和RBD重叠多达约1个氨基酸、多达约5个氨基酸、多达约10个氨基酸或多达约20个氨基酸。

在实施方案中，如本申请所提供的NTD可以在C末端延伸多达5、多达10、多达15、多达20、多达25或多达30个氨基酸。

在实施方案中，本申请所述的CoV S多肽包含与SEQ ID NO:1或SEQ ID NO:2的NTD具有至少95％、至少96％、至少97％、至少98％、至少99％或至少99.5％同一性的NTD。所述NTD可以具有与SEQ ID NO:1或SEQ ID NO:2的NTD的氨基酸序列相比多达约1、多达约2、多达约3、多达约4、多达约5、多达约10、多达约15、多达约20、多达约25或多达约30个氨基酸的缺失、插入或突变。所述NTD可以具有与SEQ ID NO:1或SEQ ID NO:2的NTD相比在约1与约5个之间的氨基酸、在约3与约10个之间的氨基酸、在约5与10个之间的氨基酸、在约8与12个之间的氨基酸、在约10与15个之间的氨基酸、在约12与17个之间的氨基酸、在约15与20个之间的氨基酸、在约18与23个之间的氨基酸、在约20与25个之间的氨基酸、在约22与约27个之间的氨基酸或在约25与30个之间的氨基酸的缺失、插入或突变。

在实施方案中，所述CoV S多肽含有从N末端结构域(NTD)中的一个或多个氨基酸(对应于SEQ ID NO:2的氨基酸1-292)的缺失。在实施方案中，所述CoV S多肽含有NTD的多达约10、20、30、40、50、60、70、80、90、100、110、120、130、140、150、160、170、180、190、200、210、220、230、240、250、260、270、280、290或292个氨基酸的缺失。

在实施方案中，所述CoV S多肽含有从NTD中的一个或多个氨基酸(对应于SEQ IDNO:2的氨基酸1-318)的缺失。在实施方案中，所述CoV S多肽含有SEQ ID NO:2的NTD的氨基酸1-318的缺失。在实施方案中，NTD的缺失增强了CoV刺突(S)多肽的蛋白质表达。在实施方案中，具有NTD缺失的CoV S多肽具有由SEQ ID NO:46、48、49、51、52和54表达的氨基酸序列。在实施方案中，具有NTD缺失的CoV S多肽由选自SEQ ID NO:47、SEQ ID NO:50和SEQ IDNO:53的分离的核酸序列编码。

在实施方案中，所述NTD可以含有表1B中所示的修饰的任何组合。修饰是关于供参考的成熟S多肽序列SEQ ID NO:2显示的。

表1B

CoV S多肽抗原-对S1亚基-RBD的修饰

在实施方案中，所述CoV S多肽含有对RBD的一个或多个修饰。

在实施方案中，所述RBD具有SEQ ID NO:126的氨基酸序列，其对应于SEQ ID NO:1的氨基酸331-527或SEQ ID NO:2的氨基酸318-514。

所述RBD的氨基酸序列(SEQ ID NO:126)示于下文：

NITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGP

在实施方案中，所述RBD具有SEQ ID NO:116的氨基酸序列，其对应于SEQ ID NO:1的氨基酸335-530或SEQ ID NO:2的氨基酸322-517。

所述RBD的氨基酸序列(SEQ ID NO:116)示于下文。

LCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKS

在实施方案中，如本申请所提供的RBD可以在N末端或C末端延伸多达1个氨基酸、多达5个氨基酸、多达10个氨基酸、多达15个氨基酸、多达20个氨基酸、多达25个氨基酸或多达30个氨基酸。

在实施方案中，本申请所述的CoV S多肽包含与SEQ ID NO:1或SEQ ID NO:2的RBD具有至少95％、至少96％、至少97％、至少98％、至少99％或至少99.5％同一性的RBD。所述RBD可以具有与SEQ ID NO:1或SEQ ID NO:2的RBD的氨基酸序列相比多达约1、多达约2、多达约3、多达约4、多达约5、多达约10、多达约15、多达约20、多达约25或多达约30个氨基酸的缺失、插入或突变。所述RBD可以具有与SEQ ID NO:1或SEQ ID NO:2的RBD相比在约1与约5个之间的氨基酸、在约3与约10个之间的氨基酸、在约5与10个之间的氨基酸、在约8与12个之间的氨基酸、在约10与15个之间的氨基酸、在约12与17个之间的氨基酸、在约15与20个之间的氨基酸、在约18与23个之间的氨基酸、在约20与25个之间的氨基酸、在约22与约27个之间的氨基酸或在约25与30个之间的氨基酸的缺失、插入或突变。

在实施方案中，所述CoV S多肽在RBD中具有至少一、至少二、至少三、至少四、至少5、至少6、至少7、至少8、至少9、至少10、至少11、至少12、至少13、至少14、至少15、至少16、至少17、至少18、至少19或至少20个突变。在实施方案中，所述RBD可以含有如表1C中所示的修饰的任何组合。

表1C

CoV S多肽抗原-对SD1/2的修饰

在实施方案中，所述CoV S多肽含有对具有SEQ ID NO:122的氨基酸序列的SD1/2结构域的一个或多个修饰，所述氨基酸序列对应于SEQ ID NO:1的氨基酸542-681或SEQ IDNO:2的氨基酸529-668。

所述SD1/2(SEQ ID NO:122)结构域的氨基酸序列示于下文。

NFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSP

在实施方案中，本申请所述的CoV S多肽包含与SEQ ID NO:1或SEQ ID NO:2的SD1/2具有至少95％、至少96％、至少97％、至少98％、至少99％或至少99.5％同一性的SD1/2结构域。所述SD1/2结构域可以具有与SEQ ID NO:1或SEQ ID NO:2的SD1/2的氨基酸序列相比多达约1、多达约2、多达约3、多达约4、多达约5、多达约10、多达约15、多达约20、多达约25或多达约30个氨基酸的缺失、插入或突变。所述SD1/2结构域可以具有与SEQ ID NO:1或SEQ ID NO:2的SD1/2结构域相比在约1与约5个之间的氨基酸、在约3与约10个之间的氨基酸、在约5与10个之间的氨基酸、在约8与12个之间的氨基酸、在约10与15个之间的氨基酸、在约12与17个之间的氨基酸、在约15与20个之间的氨基酸、在约18与23个之间的氨基酸、在约20与25个之间的氨基酸、在约22与约27个之间的氨基酸或在约25与30个之间的氨基酸的缺失、插入或突变。

在实施方案中，所述CoV S多肽在SD1/2结构域中具有至少一、至少二、至少三、至少四、至少5、至少6、至少7、至少8、至少9、至少10、至少11、至少12、至少13、至少14、至少15、至少16、至少17、至少18、至少19或至少20个突变。在实施方案中，所述SD1/2结构域可以含有如表1D中所示的修饰的任何组合。

表1D

CoV S多肽抗原-对弗林蛋白酶切割位点的修饰

在实施方案中，所述CoV S多肽含有被一个或多个突变灭活的弗林蛋白酶位点(RRAR)，其对应于SEQ ID NO:1的氨基酸682-685或SEQ ID NO:2的氨基酸669-672。弗林蛋白酶切割位点的灭活防止弗林蛋白酶切割CoV S多肽。在实施方案中，本申请所述的含有灭活的弗林蛋白酶切割位点的CoV S多肽被表达为单链。

在实施方案中，构成天然弗林蛋白酶切割位点的一个或多个氨基酸突变为任何天然氨基酸。在实施方案中，所述氨基酸是L-氨基酸。氨基酸的非限制性例子包括丙氨酸、精氨酸、甘氨酸、天门冬酰胺、天门冬氨酸、半胱氨酸、谷氨酰胺、谷氨酸、丝氨酸、苏氨酸、组氨酸、赖氨酸、甲硫氨酸、脯氨酸、缬氨酸、异亮氨酸、酪氨酸、色氨酸和苯丙氨酸。

在实施方案中，构成天然弗林蛋白酶切割位点的一个或多个氨基酸突变为谷氨酰胺。在实施方案中，1、2、3或4个氨基酸可以突变为谷氨酰胺。在实施方案中，构成天然弗林蛋白酶切割位点的一个精氨酸突变为谷氨酰胺。在实施方案中，构成天然弗林蛋白酶切割位点的两个精氨酸突变为谷氨酰胺。在实施方案中，构成天然弗林蛋白酶切割位点的三个精氨酸突变为谷氨酰胺。

在实施方案中，构成天然弗林蛋白酶切割位点的一个或多个氨基酸突变为丙氨酸。在实施方案中，1、2、3或4个氨基酸可以突变为丙氨酸。在实施方案中，构成天然弗林蛋白酶切割位点的精氨酸之一突变为丙氨酸。在实施方案中，构成天然弗林蛋白酶切割位点的两个精氨酸突变为丙氨酸。在实施方案中，构成天然弗林蛋白酶切割位点的三个精氨酸突变为丙氨酸。

在实施方案中，天然弗林蛋白酶切割位点中的一个或多个氨基酸突变为甘氨酸。在实施方案中，1、2、3或4个氨基酸可以突变为甘氨酸。在实施方案中，天然弗林蛋白酶切割位点的一个精氨酸突变为甘氨酸。在实施方案中，天然弗林蛋白酶切割位点中的两个精氨酸突变为甘氨酸。在实施方案中，构成天然弗林蛋白酶切割位点的三个精氨酸突变为甘氨酸。

在实施方案中，天然弗林蛋白酶切割位点中的一个或多个氨基酸突变为天门冬酰胺。例如，1、2、3或4个氨基酸可以突变为天门冬酰胺。在实施方案中，天然弗林蛋白酶切割位点中的一个精氨酸突变为天门冬酰胺。在实施方案中，天然弗林蛋白酶切割位点中的两个精氨酸突变为天门冬酰胺。在实施方案中，天然弗林蛋白酶切割位点中的三个精氨酸突变为天门冬酰胺。

包含在所述CoV S多肽内的灭活的弗林蛋白酶位点的氨基酸序列的非限制性例子可见于表1E。

表1E

弗林蛋白酶切割位点的氨基酸序列	有活性或无活性的弗林蛋白酶切割位点
		RRAR(SEQ ID NO:6)	有活性
QQAQ(SEQ ID NO:7)	无活性
		QRAR(SEQ ID NO:8)	无活性
RQAR(SEQ ID NO:9)	无活性
		RRAQ(SEQ ID NO:10)	无活性
QQAR(SEQ ID NO:11)	无活性
		RQAQ(SEQ ID NO:12)	无活性
QRAQ(SEQ ID NO:13)	无活性
		NNAN(SEQ ID NO:14)	无活性
NRAR(SEQ ID NO:15)	无活性
		RNAR(SEQ ID NO:16)	无活性
RRAN(SEQ ID NO:17)	无活性
		NNAR(SEQ ID NO:18)	无活性
RNAN(SEQ ID NO:19)	无活性
		NRAN(SEQ ID NO:20)	无活性
AAAA(SEQ ID NO:21)	无活性
		ARAR(SEQ ID NO:22)	无活性
RAAR(SEQ ID NO:23)	无活性
		RRAA(SEQ ID NO:24)	无活性
AAAR(SEQ ID NO:25)	无活性
		RAAA(SEQ ID NO:26)	无活性
ARAA(SEQ ID NO:27)	无活性
		GGAG(SEQ ID NO:28)	无活性
GRAR(SEQ ID NO:29)	无活性
		RGAR(SEQ ID NO:30)	无活性
RRAG(SEQ ID NO:31)	无活性
		GGAR(SEQ ID NO:32)	无活性
RGAG(SEQ ID NO:33)	无活性
		GRAG(SEQ ID NO:34)	无活性
GSAS(SEQ ID NO:97)	无活性
		GSGA(SEQ ID NO:111)	无活性

在实施方案中，代替有活性的弗林蛋白酶切割位点(SEQ ID NO:6)，本申请所述的CoV S多肽含有灭活的弗林蛋白酶切割位点。在实施方案中，灭活的弗林蛋白酶切割位点的氨基酸序列由SEQ ID NO:7-34或SEQ ID NO:97中的任一个表示。在实施方案中，灭活的弗林蛋白酶切割位点的氨基酸序列为QQAQ(SEQ ID NO:7)。在实施方案中，灭活的弗林蛋白酶切割位点的氨基酸序列为GSAS(SEQ ID NO:97)。在实施方案中，灭活的弗林蛋白酶切割位点的氨基酸序列为GSGA(SEQ ID NO:111)。在实施方案中，灭活的弗林蛋白酶切割位点的氨基酸序列是GG、GGG(SEQ ID NO:127)、GGGG(SEQ ID NO:128)或GGGGG(SEQ ID NO:129)。

CoV S多肽抗原-对S2亚基的修饰

在实施方案中，所述CoV S多肽含有对具有SEQ ID NO:120的氨基酸序列的S2亚基的一个或多个修饰，所述氨基酸序列对应于SEQ ID NO:1的氨基酸686-1273或SEQ ID NO:2的氨基酸673-1260。

所述S2亚基的氨基酸序列(SEQ ID NO:120)示于下文。

SVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDKVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWYIWLGFIAGLIAIVMVTIMLCCMTSCCSCLKGCCSCGSCCKFDEDDSEPVLKGVKLHYT

在实施方案中，本申请所述的CoV S多肽包含与SEQ ID NO:1或SEQ ID NO:2的S2亚基具有至少95％、至少96％、至少97％、至少98％、至少99％或至少99.5％同一性的S2亚基。所述S2亚基可以具有与SEQ ID NO:1或SEQ ID NO:2的S2亚基的氨基酸序列相比多达约1、多达约2、多达约3、多达约4、多达约5、多达约10、多达约15、多达约20、多达约25或多达约30个氨基酸的缺失、插入或突变。所述S2亚基可以具有与SEQ ID NO:1或SEQ ID NO:2的S2亚基相比在约1与约5个之间的氨基酸、在约3与约10个之间的氨基酸、在约5与10个之间的氨基酸、在约8与12个之间的氨基酸、在约10与15个之间的氨基酸、在约12与17个之间的氨基酸、在约15与20个之间的氨基酸、在约18与23个之间的氨基酸、在约20与25个之间的氨基酸、在约22与约27个之间的氨基酸或在约25与30个之间的氨基酸的缺失、插入或突变。

在实施方案中，所述S2亚基可以含有如表1F中所示的修饰的任何组合。

表1F

在实施方案中，所述CoV S多肽含有缺失，其对应于天然CoV刺突(S)多肽(SEQ IDNO:2)的氨基酸676-685内的一个或多个缺失。在实施方案中，天然CoV刺突(S)多肽(SEQ IDNO:2)的氨基酸676-685的1、2、3、4、5、6、7、8、9或10个氨基酸缺失。在实施方案中，氨基酸676-685内的氨基酸的缺失是连续的，例如氨基酸676和677缺失或氨基酸680和681缺失。在实施方案中，氨基酸676-685内的氨基酸的缺失是非连续的，例如氨基酸676和680缺失或氨基酸677和682缺失。在实施方案中，含有对应于氨基酸676-685内的一个或多个缺失的缺失的CoV S多肽具有选自SEQ ID NO:62和SEQ ID NO:63的氨基酸序列。

在实施方案中，所述CoV S多肽含有缺失，其对应于天然CoV刺突(S)多肽(SEQ IDNO:2)的氨基酸702-711内的一个或多个缺失。在实施方案中，天然SARS-CoV-2刺突(S)多肽(SEQ ID NO:2)的氨基酸702-711的1、2、3、4、5、6、7、8、9或10个氨基酸缺失。在实施方案中，氨基酸702-711内的氨基酸的一个或多个缺失是连续的，例如氨基酸702和703缺失或氨基酸708和709缺失。在实施方案中，氨基酸702-711内的氨基酸的缺失是非连续的，例如氨基酸702和704缺失或氨基酸707和710缺失。在实施方案中，含有对应于氨基酸702-711内的一个或多个缺失的缺失的CoV S多肽具有选自SEQ ID NO:64和SEQ ID NO:65的氨基酸序列。

在实施方案中，所述CoV S多肽含有缺失，其对应于天然CoV S多肽(SEQ ID NO:2)的氨基酸775-793内的一个或多个缺失。在实施方案中，天然SARS-CoV-2刺突(S)多肽(SEQID NO:2)的氨基酸775-793的多达约1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18或19个氨基酸缺失。在实施方案中，氨基酸775-793内的氨基酸的一个或多个缺失是连续的，例如氨基酸776和777缺失或氨基酸780和781缺失。在实施方案中，氨基酸775-793内的氨基酸的缺失是非连续的，例如氨基酸775和790缺失或氨基酸777和781缺失。

在实施方案中，所述CoV S多肽含有融合肽(SEQ ID NO:104)的缺失，所述融合肽对应于SEQ ID NO:2的氨基酸806-815。在实施方案中，CoV刺突(S)多肽(SEQ ID NO:2)的融合肽的1、2、3、4、5、6、7、8、9或10个氨基酸缺失。在实施方案中，融合肽内的氨基酸的缺失是连续的，例如氨基酸806和807缺失或氨基酸809和810缺失。在实施方案中，融合肽内的氨基酸的缺失是非连续的，例如氨基酸806和808缺失或氨基酸810和813缺失。在实施方案中，含有对应于融合肽的一个或多个氨基酸的缺失的CoV S多肽具有选自SEQ ID NO:66、77和105-108的氨基酸序列。

在实施方案中，所述CoV S多肽在天然CoV刺突(S)多肽(SEQ ID NO:2)的Lys-973处含有突变。在实施方案中，Lys-973突变为任何天然氨基酸。在实施方案中，Lys-973突变为脯氨酸。在实施方案中，Lys-973突变为甘氨酸。在实施方案中，含有在氨基酸973处的突变的CoV S多肽选自SEQ ID NO:84-89、105-106和109-110。

在实施方案中，所述CoV S多肽含有在天然CoV刺突(S)多肽(SEQ ID NO:2)的Val-974处的突变。在实施方案中，Val-974突变为任何天然氨基酸。在实施方案中，Val-974突变为脯氨酸。在实施方案中，Val-974突变为甘氨酸。在实施方案中，含有在氨基酸974处的突变的CoV S多肽选自SEQ ID NO:84-89、105-106和109-110。

在实施方案中，所述CoV S多肽含有在天然CoV刺突(S)多肽(SEQ ID NO:2)的Lys-973和Val-974处的突变。在实施方案中，Lys-973和Val-974突变为任何天然氨基酸。在实施方案中，Lys-973和Val-974突变为脯氨酸。在实施方案中，含有在氨基酸973和974处的突变的CoV S多肽选自SEQ ID NO:84-89、105-106和109-110。

CoV S多肽抗原-对S2亚基-HR1结构域的修饰

在实施方案中，所述CoV S多肽含有对具有SEQ ID NO:119的氨基酸序列的HR1结构域的一个或多个修饰，所述氨基酸序列对应于SEQ ID NO:1的氨基酸912-984或SEQ IDNO:2的氨基酸889-971。

所述HR1结构域的氨基酸序列(SEQ ID NO:119)示于下文。

MAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRL

在实施方案中，本申请所述的CoV S多肽包含与SEQ ID NO:1或SEQ ID NO:2的HR1结构域具有至少95％、至少96％、至少97％、至少98％、至少99％或至少99.5％同一性的HR1结构域。所述HR1结构域可以具有与SEQ ID NO:1或SEQ ID NO:2的HR1结构域的氨基酸序列相比多达约1、多达约2、多达约3、多达约4、多达约5、多达约10、多达约15、多达约20、多达约25或多达约30个氨基酸的缺失、插入或突变。所述HR1结构域可以具有与SEQ ID NO:1或SEQID NO:2的HR1结构域相比在约1与约5个之间的氨基酸、在约3与约10个之间的氨基酸、在约5与10个之间的氨基酸、在约8与12个之间的氨基酸、在约10与15个之间的氨基酸、在约12与17个之间的氨基酸、在约15与20个之间的氨基酸、在约18与23个之间的氨基酸、在约20与25个之间的氨基酸、在约22与约27个之间的氨基酸或在约25与30个之间的氨基酸的缺失、插入或突变。

在实施方案中，所述HR1结构域可以含有如表1G中所示的修饰的任何组合。

表1G

CoV S多肽抗原-对S2亚基-HR2结构域的修饰

在实施方案中，所述CoV S多肽含有对具有SEQ ID NO:125的氨基酸序列的HR2结构域的一个或多个修饰，所述氨基酸序列对应于SEQ ID NO:1的氨基酸1163-1213或SEQ IDNO:2的氨基酸1150-1200。

所述HR2结构域的氨基酸序列(SEQ ID NO:125)示于下文。

DVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWP

在实施方案中，本申请所述的CoV S多肽包含与SEQ ID NO:1或SEQ ID NO:2的HR2结构域具有至少95％、至少96％、至少97％、至少98％、至少99％或至少99.5％同一性的HR2结构域。所述HR2结构域可以具有与SEQ ID NO:1或SEQ ID NO:2的HR2结构域的氨基酸序列相比多达约1、多达约2、多达约3、多达约4、多达约5、多达约10、多达约15、多达约20、多达约25或多达约30个氨基酸的缺失、插入或突变。所述HR2结构域可以具有与SEQ ID NO:1或SEQID NO:2的HR2结构域相比在约1与约5个之间的氨基酸、在约3与约10个之间的氨基酸、在约5与10个之间的氨基酸、在约8与12个之间的氨基酸、在约10与15个之间的氨基酸、在约12与17个之间的氨基酸、在约15与20个之间的氨基酸、在约18与23个之间的氨基酸、在约20与25个之间的氨基酸、在约22与约27个之间的氨基酸或在约25与30个之间的氨基酸的缺失、插入或突变。

CoV S多肽抗原-对TM结构域的修饰

在实施方案中，所述CoV S多肽含有对具有SEQ ID NO:123的氨基酸序列的TM结构域的一个或多个修饰，所述氨基酸序列对应于SEQ ID NO:1的氨基酸1214-1237或SEQ IDNO:2的氨基酸1201-1224。

所述TM结构域的氨基酸序列(SEQ ID NO:123)示于下文。

WYIWLGFIAGLIAIVMVTIMLCCM

在实施方案中，本申请所述的CoV S多肽包含与SEQ ID NO:1或SEQ ID NO:2的TM结构域具有至少95％、至少96％、至少97％、至少98％、至少99％或至少99.5％同一性的TM结构域。所述TM结构域可以具有与SEQ ID NO:1或SEQ ID NO:2的TM结构域的氨基酸序列相比多达约1、多达约2、多达约3、多达约4、多达约5、多达约10、多达约15、多达约20、多达约25或多达约30个氨基酸的缺失、插入或突变。所述TM结构域可以具有与SEQ ID NO:1或SEQ IDNO:2的TM结构域相比在约1与约5个之间的氨基酸、在约3与约10个之间的氨基酸、在约5与10个之间的氨基酸、在约8与12个之间的氨基酸、在约10与15个之间的氨基酸、在约12与17个之间的氨基酸、在约15与20个之间的氨基酸、在约18与23个之间的氨基酸、在约20与25个之间的氨基酸、在约22与约27个之间的氨基酸或在约25与30个之间的氨基酸的缺失、插入或突变。

在实施方案中，本申请所述的CoV S多肽缺少整个TM结构域。在实施方案中，所述CoV S多肽包含TM结构域。

CoV S多肽抗原-对CT的修饰

在实施方案中，所述CoV S多肽含有对具有SEQ ID NO:124的氨基酸序列的CT的一个或多个修饰，所述氨基酸序列对应于SEQ ID NO:1的氨基酸1238-1273或SEQ ID NO:2的氨基酸1225-1260。

所述CT的氨基酸序列(SEQ ID NO:124)示于下文：

TSCCSCLKGCCSCGSCCKFDEDDSEPVLKGVKLHYT

在实施方案中，本申请所述的CoV S多肽包含与SEQ ID NO:1或SEQ ID NO:2的CT具有至少95％、至少96％、至少97％、至少98％、至少99％或至少99.5％同一性的CT。所述CT可以具有与SEQ ID NO:1或SEQ ID NO:2的CT的氨基酸序列相比多达约1、多达约2、多达约3、多达约4、多达约5、多达约10、多达约15、多达约20、多达约25或多达约30个氨基酸的缺失、插入或突变。所述CT可以具有与SEQ ID NO:1或SEQ ID NO:2的CT相比在约1与约5个之间的氨基酸、在约3与约10个之间的氨基酸、在约5与10个之间的氨基酸、在约8与12个之间的氨基酸、在约10与15个之间的氨基酸、在约12与17个之间的氨基酸、在约15与20个之间的氨基酸、在约18与23个之间的氨基酸、在约20与25个之间的氨基酸、在约22与约27个之间的氨基酸或在约25与30个之间的氨基酸的缺失、插入或突变。

在实施方案中，本申请所述的CoV S多肽缺少CT。在实施方案中，所述CoV S多肽包含CT。

在实施方案中，所述CoV S多肽包含TM和CT。在实施方案中，所述CoV刺突(S)多肽含有从跨膜和胞质尾(TMCT)(对应于氨基酸1201-1260)中的一个或多个氨基酸的缺失。所述TMCT的氨基酸序列由SEQ ID NO:39表示。在实施方案中，具有TMCT的一个或多个残基的缺失的CoV S多肽具有增强的蛋白质表达。在实施方案中，具有来自TMCT的一个或多个缺失的CoV刺突(S)多肽具有选自SEQ ID NO:40、41、42、52、54、59、61、88和89的氨基酸序列。在实施方案中，具有来自TM-CD的一个或多个缺失的CoV S多肽由选自SEQ ID NO:39、43、53和60的分离的核酸序列编码。

CoV S多肽抗原-突变的非限制性组合

在实施方案中，所述CoV S多肽含有天然CoV刺突(S)多肽(SEQ ID NO:2)的氨基酸56和57的缺失。

在实施方案中，所述CoV S多肽含有天然CoV刺突(S)多肽(SEQ ID NO:2)的氨基酸131和132的缺失。

在实施方案中，所述CoV S多肽含有天然CoV刺突(S)多肽(SEQ ID NO:2)的氨基酸56和131的缺失。在实施方案中，所述CoV S多肽含有天然CoV刺突(S)多肽(SEQ ID NO:2)的氨基酸57和131的缺失。

在实施方案中，所述CoV S多肽含有天然CoV刺突(S)多肽(SEQ ID NO:2)的氨基酸56、57和131的缺失。

在实施方案中，所述CoV S多肽含有天然CoV刺突(S)多肽(SEQ ID NO:2)的氨基酸56和132的缺失。

在实施方案中，所述CoV S多肽含有天然CoV刺突(S)多肽(SEQ ID NO:2)的氨基酸57和132的缺失。

在实施方案中，所述CoV S多肽含有天然CoV刺突(S)多肽(SEQ ID NO:2)的氨基酸56、57和132的缺失。

在实施方案中，所述CoV S多肽含有天然CoV刺突(S)多肽(SEQ ID NO:2)的氨基酸56、57、131和132的缺失。

在实施方案中，所述CoV S多肽含有使CoV S多肽的融合前构象稳定的突变。在实施方案中，所述CoV S多肽含有使融合前构象稳定的脯氨酸或甘氨酸取代。该策略已用于开发融合前稳定化的MERS-CoV S蛋白，如以下文档中所述，这些文档的每一个均通过引用以其整体并入本申请：Proc Natl Acad Sci USA.2017年8月29日；114(35):E7348-E7357；SciRep.2018年10月24日；8(1):15701；美国公开号2020/0061185；以及PCT申请号PCT/US2017/058370。

在实施方案中，所述CoV S多肽含有在Lys-973和Val-974处的突变以及灭活的弗林蛋白酶切割位点。在实施方案中，所述CoV S多肽含有Lys-973和Val-974至脯氨酸的突变以及具有QQAQ(SEQ ID NO:7)或GSAS(SEQ ID NO:96)的氨基酸序列的灭活的弗林蛋白酶切割位点。含有在Lys-973和Val-974处的突变以及灭活的弗林蛋白酶切割位点的示例性CoVS多肽描绘在图8中。在实施方案中，含有Lys-973和Val-974至脯氨酸的突变以及灭活的弗林蛋白酶切割位点的CoV S多肽具有SEQ ID NO:86或87的氨基酸序列和SEQ ID NO:96的核酸序列。

在实施方案中，所述CoV S多肽含有在Lys-973和Val-974处的突变、灭活的弗林蛋白酶切割位点以及融合肽的一个或多个氨基酸的缺失。在实施方案中，所述CoV S多肽含有Lys-973和Val-974至脯氨酸的突变、具有QQAQ(SEQ ID NO:7)或GSAS(SEQ ID NO:96)的氨基酸序列的灭活的弗林蛋白酶切割位点以及融合肽的一个或多个氨基酸的缺失。在实施方案中，含有Lys-973和Val-974至脯氨酸的突变、灭活的弗林蛋白酶切割位点以及融合肽的一个或多个氨基酸的缺失的CoV S多肽具有SEQ ID NO:105或106的氨基酸序列。在实施方案中，相对于天然CoV刺突(S)多肽(SEQ ID NO:2)，所述CoV S多肽含有Leu-5至苯丙氨酸的突变、Thr-7至天门冬酰胺的突变、Pro-13至丝氨酸的突变、Asp-125至酪氨酸的突变、Arg-177至丝氨酸的突变、Lys-404至苏氨酸的突变、Glu-471至赖氨酸的突变、Asn-488至酪氨酸的突变、His-642至酪氨酸的突变、Thr-1014至异亮氨酸的突变、Lys-973和Val-974至脯氨酸的突变以及具有QQAQ(SEQ ID NO:7)或GSAS(SEQ ID NO:96)的氨基酸序列的灭活的弗林蛋白酶切割位点。

在实施方案中，相对于天然CoV刺突(S)多肽(SEQ ID NO:2)，所述CoV S多肽含有Trp-139至半胱氨酸的突变、Leu-439至精氨酸的突变、Lys-973和Val-974至脯氨酸的突变以及具有QQAQ(SEQ ID NO:7)或GSAS(SEQ ID NO:96)的氨基酸序列的灭活的弗林蛋白酶切割位点。在实施方案中，相对于天然CoV刺突(S)多肽(SEQ ID NO:1)，所述CoV S多肽含有Trp-152至半胱氨酸的突变、Leu-452至精氨酸的突变、Ser-13至异亮氨酸的突变、Lys-986和Val-987至脯氨酸的突变以及具有QQAQ(SEQ ID NO:7)或GSAS(SEQ ID NO:96)的氨基酸序列的灭活的弗林蛋白酶切割位点。

在实施方案中，相对于天然CoV刺突(S)多肽(SEQ ID NO:2)，所述CoV S多肽含有Lys-404至苏氨酸或天门冬酰胺的突变、Glu-471至赖氨酸的突变、Asn-488至酪氨酸的突变、Leu-5至苯丙氨酸的突变、Asp-67至丙氨酸的突变、Asp-202至甘氨酸的突变、氨基酸229-231中的一个或多个的缺失、Arg-233至异亮氨酸的突变、Lys-973和Val-974至脯氨酸的突变以及具有QQAQ(SEQ ID NO:7)或GSAS(SEQ ID NO:96)的氨基酸序列的灭活的弗林蛋白酶切割位点。

在实施方案中，相对于天然CoV刺突(S)多肽(SEQ ID NO:2)，所述CoV S多肽含有Asn-488至酪氨酸的突变、Lys-973和Val-974至脯氨酸的突变以及具有QQAQ(SEQ ID NO:7)或GSAS(SEQ ID NO:96)的氨基酸序列的灭活的弗林蛋白酶切割位点。在实施方案中，具有Asn-488至酪氨酸的突变、Lys-973和Val-974至脯氨酸的突变以及具有QQAQ(SEQ ID NO:7)或GSAS(SEQ ID NO:96)的氨基酸序列的灭活的弗林蛋白酶切割位点的CoV S多肽包含SEQID NO:112的氨基酸序列。

在实施方案中，相对于天然CoV刺突(S)多肽(SEQ ID NO:2)，所述CoV S多肽含有Asp-601至甘氨酸的突变、Asn-488至酪氨酸的突变、Lys-973和Val-974至脯氨酸的突变以及具有QQAQ(SEQ ID NO:7)或GSAS(SEQ ID NO:96)的氨基酸序列的灭活的弗林蛋白酶切割位点。在实施方案中，具有Asn-488至酪氨酸的突变、Lys-973和Val-974至脯氨酸的突变以及具有QQAQ(SEQ ID NO:7)或GSAS(SEQ ID NO:96)的氨基酸序列的灭活的弗林蛋白酶切割位点的CoV S多肽包含SEQ ID NO:113的氨基酸序列。

在实施方案中，相对于天然CoV刺突(S)多肽(SEQ ID NO:2)，所述CoV S多肽含有氨基酸56、57和131的缺失、Asn-488至酪氨酸的突变、Ala-557至天门冬氨酸的突变、Asp-601至甘氨酸的突变、Pro-668至组氨酸的突变、Thr-703至异亮氨酸的突变、Ser-969至丙氨酸的突变、Asp-1105至组氨酸的突变、Lys-973和Val-974至脯氨酸的突变以及具有QQAQ(SEQ ID NO:7)、GSAS(SEQ ID NO:96)或GG的氨基酸序列的灭活的弗林蛋白酶切割位点。在实施方案中，具有氨基酸56、57和131的缺失、Asn-488至酪氨酸的突变、Ala-557至天门冬氨酸的突变、Asp-601至甘氨酸的突变、Pro-668至组氨酸的突变、Thr-703至异亮氨酸的突变、Ser-969至丙氨酸的突变、Asp-1105至组氨酸的突变、Lys-973和Val-974至脯氨酸的突变以及具有QQAQ(SEQ ID NO:7)或GSAS(SEQ ID NO:96)的氨基酸序列的灭活的弗林蛋白酶切割位点的CoV S多肽包含SEQ ID NO:114的氨基酸序列。在实施方案中，具有氨基酸56、57和131的缺失、Asn-488至酪氨酸的突变、Ala-557至天门冬氨酸的突变、Asp-601至甘氨酸的突变、Pro-668至组氨酸的突变、Thr-703至异亮氨酸的突变、Ser-969至丙氨酸的突变、Asp-1105至组氨酸的突变、Lys-973和Val-974至脯氨酸的突变以及具有QQAQ(SEQ ID NO:7)或GSAS(SEQ ID NO:96)或GG的氨基酸序列的灭活的弗林蛋白酶切割位点的CoV S多肽包含SEQ ID NO:136的氨基酸序列。在实施方案中，具有氨基酸56、57和131的缺失、Asn-488至酪氨酸的突变、Ala-557至天门冬氨酸的突变、Asp-601至甘氨酸的突变、Pro-668至组氨酸的突变、Thr-703至异亮氨酸的突变、Ser-969至丙氨酸的突变、Asp-1105至组氨酸的突变、Lys-973和Val-974至脯氨酸的突变以及具有GG的氨基酸序列的灭活的弗林蛋白酶切割位点的CoV S多肽包含SEQ ID NO:137或SEQ ID NO:138的氨基酸序列。在一些实施方案中，具有SEQ ID NO:114或SEQ ID NO:136的氨基酸序列的CoV S多肽由具有SEQ ID NO:135的核酸序列的核酸编码。在一些实施方案中，具有SEQ ID NO:137或SEQ ID NO:138的氨基酸序列的CoV S多肽由具有SEQ ID NO:139的序列的核酸编码。

在实施方案中，相对于天然CoV刺突(S)多肽(SEQ ID NO:2)，所述CoV S多肽含有氨基酸56、57和132的缺失、Asn-488至酪氨酸的突变、Ala-557至天门冬氨酸的突变、Asp-601至甘氨酸的突变、Pro-668至组氨酸的突变、Thr-703至异亮氨酸的突变、Ser-969至丙氨酸的突变、Asp-1105至组氨酸的突变、Lys-973和Val-974至脯氨酸的突变以及具有QQAQ(SEQ ID NO:7)或GSAS(SEQ ID NO:96)的氨基酸序列的灭活的弗林蛋白酶切割位点。在实施方案中，具有氨基酸56、57和132的缺失、Asn-488至酪氨酸的突变、Ala-557至天门冬氨酸的突变、Asp-601至甘氨酸的突变、Pro-668至组氨酸的突变、Thr-703至异亮氨酸的突变、Ser-969至丙氨酸的突变、Asp-1105至组氨酸的突变、Lys-973和Val-974至脯氨酸的突变以及具有QQAQ(SEQ ID NO:7)或GSAS(SEQ ID NO:96)的氨基酸序列的灭活的弗林蛋白酶切割位点的CoV S多肽包含SEQ ID NO:114的氨基酸序列。

在实施方案中，相对于天然CoV刺突(S)多肽(SEQ ID NO:2)，所述CoV S多肽含有Asn-488至酪氨酸的突变、Asp-67至丙氨酸的突变、Leu-229至组氨酸的突变、Asp-202至甘氨酸的突变、Lys-404至天门冬酰胺的突变、Glu-471至赖氨酸的突变、Ala-688至缬氨酸的突变、Asp-601至甘氨酸的突变、Lys-973和Val-974至脯氨酸的突变以及具有QQAQ(SEQ IDNO:7)或GSAS(SEQ ID NO:96)的氨基酸序列的灭活的弗林蛋白酶切割位点。在实施方案中，具有Asn-488至酪氨酸的突变、Asp-67至丙氨酸的突变、Leu-229至组氨酸的突变、Asp-202至甘氨酸的突变、Lys-404至天门冬酰胺的突变、Glu-471至赖氨酸的突变、Ala-688至缬氨酸的突变、Asp-601至甘氨酸的突变、Lys-973和Val-974至脯氨酸的突变以及具有QQAQ(SEQID NO:7)或GSAS(SEQ ID NO:96)的氨基酸序列的灭活的弗林蛋白酶切割位点的CoV S多肽包含SEQ ID NO:115的氨基酸序列。

在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、灭活的弗林蛋白酶切割位点、氨基酸56的缺失、氨基酸57的缺失、氨基酸131的缺失、N488Y、A557D、D601G、P668H、T703I、S969A和D1105H，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，所述灭活的弗林蛋白酶切割位点具有QQAQ(SEQ ID NO:7)的氨基酸序列。在实施方案中，所述灭活的弗林蛋白酶切割位点具有GG的氨基酸序列。

在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、灭活的弗林蛋白酶切割位点、D67A、D202G、L229H、K404N、E471K、N488Y、D601G和A688V，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，所述灭活的弗林蛋白酶切割位点具有QQAQ(SEQ ID NO:7)的氨基酸序列。在实施方案中，所述灭活的弗林蛋白酶切割位点具有GG的氨基酸序列。

在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、灭活的弗林蛋白酶切割位点、氨基酸229-231的缺失、D67A、D202G、K404N、E471K、N488Y、D601G和A688V，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。

在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、具有QQAQ(SEQ ID NO:7)的氨基酸序列的灭活的弗林蛋白酶切割位点、氨基酸229-231的缺失、L5F、D67A、D202G、K404N、E471K、N488Y、D601G和A688V，其中所述氨基酸关于具有SEQ IDNO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，具有选自以下的一个或多个修饰的CoV S多肽包含SEQ ID NO:144的氨基酸序列：K973P、V974P、具有QQAQ(SEQ ID NO:7)的氨基酸序列的灭活的弗林蛋白酶切割位点、氨基酸229-231的缺失、L5F、D67A、D202G、K404N、E471K、N488Y、D601G和A688V，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，具有SEQ ID NO:144的氨基酸序列的CoV S多肽由具有SEQ ID NO:145的序列的核酸编码。

在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、具有GG的氨基酸序列的灭活的弗林蛋白酶切割位点、氨基酸229-231的缺失、L5F、D67A、D202G、K404N、E471K、N488Y、D601G和A688V，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，具有选自以下的一个或多个修饰的CoV S多肽包含SEQ ID NO:144的氨基酸序列：K973P、V974P、具有GG的氨基酸序列的灭活的弗林蛋白酶切割位点、氨基酸229-231的缺失、L5F、D67A、D202G、K404N、E471K、N488Y、D601G和A688V，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，具有SEQ ID NO:144的氨基酸序列的CoV S多肽由具有SEQ ID NO:145的序列的核酸编码。

在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、灭活的弗林蛋白酶切割位点、L5F、T7N、P13S、D125Y、R177S、K404T、E471K、N488Y、D601G、H642Y、T1014I和V1163F，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，含有选自以下的一个或多个修饰的CoV S多肽具有SEQ ID NO:151的氨基酸序列：K973P、V974P、灭活的弗林蛋白酶切割位点、L5F、T7N、P13S、D125Y、R177S、K404T、E471K、N488Y、D601G、H642Y、T1014I和V1163F，其中所述氨基酸关于具有SEQID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，具有SEQ ID NO:151的氨基酸序列的CoV S多肽由具有SEQ ID NO:150的序列的核酸编码。

在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、灭活的弗林蛋白酶切割位点、氨基酸229-231的缺失、L5F、D67A、D202G、L229H、K404N、E471K、N488Y、D601G和A688V，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。

在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、灭活的弗林蛋白酶切割位点、K404N、E471K、N488Y、L5F、D67A、D202G、L229H、D601G、A688V以及氨基酸229-231的缺失，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，所述灭活的弗林蛋白酶切割位点具有QQAQ(SEQ ID NO:7)的氨基酸序列。在实施方案中，所述灭活的弗林蛋白酶切割位点具有GG的氨基酸序列。

在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、灭活的弗林蛋白酶切割位点、K404N、E471K和N488K，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、灭活的弗林蛋白酶切割位点、K404N、E471K和N488Y。在实施方案中，所述CoV S多肽是具有选自以下的一个或多个修饰的CoV S多肽的RBD：K973P、V974P、灭活的弗林蛋白酶切割位点、K404N、E471K和N488K，其中所述氨基酸关于具有SEQ ID NO 2的氨基酸序列的CoV S多肽进行编号。在实施方案中，所述CoV S多肽是具有选自以下的一个或多个修饰的CoV S多肽的RBD：K973P、V974P、灭活的弗林蛋白酶切割位点、K404N、E471K和N488Y，其中所述氨基酸关于具有SEQ ID NO 2的氨基酸序列的CoV S多肽进行编号。

在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、具有GG的氨基酸序列的灭活的弗林蛋白酶切割位点、D601G、E404N、E471K和N488Y。在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、具有GG的氨基酸序列的灭活的弗林蛋白酶切割位点和D601G突变，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，含有选自以下的修饰的CoV S多肽具有SEQID NO:133的氨基酸序列：K973P、V974P、具有GG氨基酸序列的灭活的弗林蛋白酶切割位点和D601G突变。

在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、灭活的弗林蛋白酶切割位点(任选地其中所述灭活的弗林蛋白酶切割位点是QQAQ(SEQ IDNO:7)或GG)、K404N、E471K、N488K、D67A、D202G、L229H、D601G和A688V，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，含有选自以下的一个或多个修饰的CoV S多肽具有SEQ ID NO:132或SEQ ID NO:141的氨基酸序列：K973P、V974P、灭活的弗林蛋白酶切割位点(任选地其中所述灭活的弗林蛋白酶切割位点是QQAQ(SEQ ID NO:7)或GG)、K404N、E471K、N488K、D67A、D202G、L229H、D601G和A688V。在实施方案中，具有SEQ ID NO:132的氨基酸序列的CoV S多肽由具有SEQ ID NO:131的核酸序列的核酸编码。在实施方案中，具有SEQ ID NO:132的氨基酸序列的CoV S多肽由具有SEQ ID NO:142的核酸序列的核酸编码。

在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、灭活的弗林蛋白酶切割位点、W139C和L439R，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，包含K973P、V974P、灭活的弗林蛋白酶切割位点、W139C和L439R修饰的CoV S多肽与具有SEQ ID NO:117或SEQ ID NO:5的氨基酸序列的信号肽一起表达。在实施方案中，所述CoV S多肽包含选自以下的一个或多个修饰：K973P、V974P、灭活的弗林蛋白酶切割位点、D601G、W139C和L439R，其中所述氨基酸关于具有SEQID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，所述CoV S多肽包含K973P、V974P、灭活的弗林蛋白酶切割位点、D601G、W139C和L439R修饰并且与具有SEQ ID NO:117或SEQ ID NO:5的氨基酸序列的信号肽一起表达。

在实施方案中，所述CoV S多肽包含选自以下的一个或多个修饰：K973P、V974P、灭活的弗林蛋白酶切割位点、D601G、L5F、D67A、D202G、氨基酸229-231的缺失、R233I、K404N、E471K、N488Y和A688V，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。

在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、灭活的弗林蛋白酶切割位点(任选地其中所述灭活的弗林蛋白酶切割位点是QQAQ(SEQ IDNO:7))、W139C、S481P、D601G和L439R，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、灭活的弗林蛋白酶切割位点(任选地其中所述灭活的弗林蛋白酶切割位点是QQAQ(SEQ ID NO:7))、W139C、D601G和L439R，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、灭活的弗林蛋白酶切割位点(任选地其中所述灭活的弗林蛋白酶切割位点是QQAQ(SEQ ID NO:7))、W139C、S481P和D601G，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，含有选自以下的一个或多个修饰的CoV S多肽具有SEQ ID NO:153的氨基酸序列：K973P、V974P、灭活的弗林蛋白酶切割位点(任选地其中所述灭活的弗林蛋白酶切割位点是QQAQ(SEQ ID NO:7))、W139C、S481P、D601G和L439R。在实施方案中，具有SEQ ID NO:153的氨基酸序列的CoV S多肽包含具有SEQ IDNO:117的氨基酸序列的信号肽。在实施方案中，具有SEQ ID NO:153的氨基酸序列的CoV S多肽包含具有SEQ ID NO:5的氨基酸序列的信号肽。

在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、灭活的弗林蛋白酶切割位点(任选地其中所述灭活的弗林蛋白酶切割位点是QQAQ(SEQ IDNO:7))、T82I、D240G、E471K、D601G和A688V，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，含有选自以下的一个或多个修饰的CoV S多肽具有SEQ ID NO:156的氨基酸序列：K973P、V974P、灭活的弗林蛋白酶切割位点(任选地其中所述灭活的弗林蛋白酶切割位点是QQAQ(SEQ ID NO:7))、T82I、D240G、E471K、D601G和A688V，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，含有选自以下的一个或多个修饰的CoV S多肽包含具有SEQ ID NO:154或SEQ IDNO:5的氨基酸序列的信号肽：K973P、V974P、灭活的弗林蛋白酶切割位点(任选地其中所述灭活的弗林蛋白酶切割位点是QQAQ(SEQ ID NO:7))、T82I、D240G、E471K、D601G和A688V，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。

在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、灭活的弗林蛋白酶切割位点(任选地其中所述灭活的弗林蛋白酶切割位点是QQAQ(SEQ IDNO:7))、T82I、D240G、S464N、D601G和A688V，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，含有选自以下的一个或多个修饰的CoV S多肽具有SEQ ID NO:158的氨基酸序列：K973P、V974P、灭活的弗林蛋白酶切割位点(任选地其中所述灭活的弗林蛋白酶切割位点是QQAQ(SEQ ID NO:7))、T82I、D240G、S464N、D601G和A688V，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，含有选自以下的一个或多个修饰的CoV S多肽包含SEQ ID NO:154的信号肽：K973P、V974P、灭活的弗林蛋白酶切割位点(任选地其中所述灭活的弗林蛋白酶切割位点是QQAQ(SEQ ID NO:7))、T82I、D240G、S464N、D601G和A688V，其中所述氨基酸关于具有SEQ IDNO:2的氨基酸序列的CoV S多肽进行编号。

在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、灭活的弗林蛋白酶切割位点(任选地其中所述灭活的弗林蛋白酶切割位点是QQAQ(SEQ IDNO:7))、氨基酸56的缺失、氨基酸57的缺失、氨基酸131的缺失、N488Y突变、A557D突变、D601G突变、P668H突变、T703I突变、S969A突变和D1105H突变，其中所述CoV S多肽关于具有SEQ ID NO:2的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、灭活的弗林蛋白酶切割位点(任选地其中所述灭活的弗林蛋白酶切割位点是QQAQ(SEQ ID NO:7))、氨基酸56的缺失、氨基酸57的缺失、氨基酸132的缺失、N488Y突变、A557D突变、D601G突变、P668H突变、T703I突变、S969A突变和D1105H突变，其中所述CoV S多肽关于具有SEQ ID NO:2的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。

在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、灭活的弗林蛋白酶切割位点(任选地其中所述灭活的弗林蛋白酶切割位点是QQAQ(SEQ IDNO:7))、D67A突变、L229H突变、R233I突变、A688V突变、N488Y突变、K404N突变、E471K突变和D601G突变，其中所述CoV S多肽关于具有SEQ ID NO:2的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。

在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K973P、V974P、灭活的弗林蛋白酶切割位点(任选地其中所述灭活的弗林蛋白酶切割位点是QQAQ(SEQ IDNO:7))、L5F突变、T7N突变、P13S突变、D125Y突变、R177S突变、K404T突变、E471K突变、N488Y突变、D601G突变、H642Y突变、T1014I突变和T1163F突变，其中所述CoV S多肽关于具有SEQID NO:2的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。

在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K986P、V987P、灭活的弗林蛋白酶切割位点(任选地其中所述灭活的弗林蛋白酶切割位点是QQAQ(SEQ IDNO:7))、S13I突变、W152C突变和L452R突变，其中所述CoV S多肽关于具有SEQ ID NO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。在实施方案中，所述CoV S多肽含有选自以下的一个或多个修饰：K986P、V987P、灭活的弗林蛋白酶切割位点(任选地其中所述灭活的弗林蛋白酶切割位点是QQAQ(SEQ ID NO:7))、S13I突变、W152C突变和L452R突变，其中所述CoV S多肽关于具有SEQ ID NO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号，缺乏N末端信号肽。

在实施方案中，所述CoV刺突(S)多肽包含多肽接头。在实施方案中，所述多肽接头含有甘氨酸和丝氨酸。在实施方案中，所述接头具有约50％、约55％、约60％、约65％、约70％、约75％、约80％、约85％、约90％、约95％或约100％甘氨酸。

在实施方案中，所述多肽接头具有重复序列(SGGG)_n(SEQ ID NO:91)，其中n是1至50的整数(例如1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、46、47、48、49或50)。在实施方案中，所述多肽接头具有对应于SEQ ID NO:90的氨基酸序列。

在实施方案中，所述多肽接头具有重复序列(GGGGS)_n(SEQ ID NO:93)，其中n是1至50的整数(例如1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、46、47、48、49或50)。

在实施方案中，所述多肽接头具有重复序列(GGGS)_n(SEQ ID NO:92)，其中n是1至50的整数(例如1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、46、47、48、49或50)。

在各个方面，所述多肽接头是聚(Gly)n接头，其中n是1、2、3、4、5、6、7、8、9、10、15、16、17、18、19或20。在其他实施方案中，所述接头选自：二肽、三肽和四肽。在实施方案中，所述接头是选自丙氨酸-丝氨酸(AS)、亮氨酸-谷氨酸(LE)和丝氨酸-精氨酸(SR)的二肽。

在实施方案中，所述多肽接头包含天然存在的CoV S多肽或本申请所公开的CoV S多肽的1至100个之间的连续氨基酸。在实施方案中，所述多肽接头具有对应于SEQ ID NO:94的氨基酸序列。

在实施方案中，所述CoV刺突(S)多肽包含折叠子。在实施方案中，所述TMCT被折叠子替代。在实施方案中，折叠子引起CoV刺突(S)多肽三聚化。在实施方案中，所述折叠子是本领域已知的氨基酸序列。在实施方案中，所述折叠子具有SEQ ID NO:68的氨基酸序列。在实施方案中，所述折叠子是T4次要纤维蛋白(fibritin)三聚化基序。在实施方案中，T4次要纤维蛋白三聚化结构域具有SEQ ID NO:103的氨基酸序列。在实施方案中，所述折叠子的氨基酸序列由多肽接头与CoV刺突(S)多肽分开。多肽接头的非限制性例子可见于整个本申请公开文本。

在实施方案中，本申请公开文本提供了包含冠状病毒S蛋白的片段的CoV S多肽以及包含所述多肽的纳米颗粒和疫苗。在实施方案中，冠状病毒S蛋白的片段的长度在10与1500个氨基酸之间的长度(例如长度为约10、约20、约30、约40、约50、约60、约70、约80、约90、约100、约150、约200、约250、约300、约350、约400、约450、约500、约550、约600、约650、约700、约750、约800、约850、约900、约950、约1000、约1050、约1100、约1150、约1200、约1250、约1300、约1350、约1400、约1450或约1500个氨基酸)。在实施方案中，冠状病毒S蛋白的片段选自受体结合结构域(RBD)、亚结构域1、亚结构域2、上螺旋、融合肽、连接区、七肽重复区1、中央螺旋、七肽重复区2、NTD和TMCT。

在实施方案中，所述CoV S多肽包含RBD和亚结构域1。在实施方案中，包含RBD和亚结构域1的CoV S多肽是SEQ ID NO:1的氨基酸319至591。

在实施方案中，所述CoV S多肽含有冠状病毒S蛋白的片段，其中所述冠状病毒S蛋白的片段是RBD。RBD的非限制性例子包括SARS-CoV-2(氨基酸序列＝SEQ ID NO:69)的RBD、SARS(氨基酸序列＝SEQ ID NO:70)的RBD和MERS(氨基酸序列＝SEQ ID NO:71)的RBD。

在实施方案中，所述CoV S多肽含有通过多肽接头连接的两个或更多个RBD。在实施方案中，所述多肽接头具有SEQ ID NO:90或SEQ ID NO:94的氨基酸序列。

在实施方案中，所述CoV S多肽含有1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19或20个RBD。

在一些实施方案中，所述CoV S多肽含有通过多肽接头连接的两个或更多个SARS-CoV-2RBD。在实施方案中，含有两个或更多个SARS-CoV-2RBD的抗原具有对应于SEQ ID NO:72-75之一的氨基酸序列。

在实施方案中，所述CoV S多肽含有SARS-CoV-2RBD和SARS RBD。在实施方案中，所述CoV S多肽包含SARS-CoV-2RBD和SARS RBD，其中每个RBD由多肽接头分开。在实施方案中，包含SARS-CoV-2RBD和SARS RBD的CoV S多肽具有选自SEQ ID NO:76-79的氨基酸序列。

在实施方案中，所述CoV S多肽含有SARS-CoV-2RBD和MERS RBD。在实施方案中，所述CoV S多肽包含SARS-CoV-2RBD和MERS RBD，其中每个RBD由多肽接头分开。

在实施方案中，所述CoV S多肽包含SARS RBD和MERS RBD。在实施方案中，所述CoVS多肽包含SARS RBD和MERS RBD，其中每个RBD由多肽接头分开。

在实施方案中，所述CoV S多肽包含SARS-CoV-2RBD、SARS RBD和MERS RBD。在实施方案中，所述CoV S多肽含有SARS-CoV-2RBD、SARS RBD和MERS RBD，其中每个RBD由多肽接头分开。在实施方案中，包含SARS-CoV-2RBD、SARS RBD和MERS RBD的CoV S多肽具有选自SEQ ID NO:80-83的氨基酸序列。

在实施方案中，本申请所述的CoV S多肽与N末端信号肽一起表达。在实施方案中，所述N末端信号肽具有SEQ ID NO:5(MFVFLVLLPLVSS)的氨基酸序列。在实施方案中，所述N末端信号肽具有SEQ ID NO:117(MFVFLVLLPLVSI)的氨基酸序列。在实施方案中，所述N末端信号肽具有SEQ ID NO:154(MFVFFVLLPLVSS)的氨基酸序列。在实施方案中，所述信号肽可以被使得能够表达CoV S蛋白的任何信号肽替代。在实施方案中，CoV S蛋白信号肽氨基酸中的一个或多个可以缺失或突变。保持起始的甲硫氨酸残基以起始表达。在实施方案中，所述CoV S多肽由选自以下的核酸序列编码：SEQ ID NO:35、SEQ ID NO:37、SEQ ID NO:95、SEQ ID NO:43、SEQ ID NO:47、SEQ ID NO:50、SEQ ID NO:53、SEQ ID NO:55、SEQ ID NO:57、SEQ ID NO:96、SEQ ID NO:60、SEQ ID NO:131、SEQ ID NO:135、SEQ ID NO:142、SEQ IDNO:145、SEQ ID NO:148和SEQ ID NO:150。在实施方案中，相对于天然CoV刺突(S)信号多肽(SEQ ID NO:5)，所述CoV S多肽的N末端信号肽包含在Ser-13处的突变。在实施方案中，Ser-13突变为任何天然氨基酸。在实施方案中，Ser-13突变为丙氨酸、甲硫氨酸、异亮氨酸、亮氨酸、苏氨酸或缬氨酸。在实施方案中，Ser-13突变为异亮氨酸。

所述CoV S蛋白在宿主细胞中表达后，N末端信号肽被切割以提供成熟CoV蛋白序列(SEQ ID NO:2、4、38、41、44、48、51、54、58、61、63、65、67、73、75、78、79、82、83、85、87、89、106、110、132、133、114、138、141、144、147、151、153、156和158、164-168)。在实施方案中，所述信号肽被宿主细胞蛋白酶切割。在方面中，可以从宿主细胞中分离全长蛋白质，随后切割信号肽。

在表达和纯化期间从具有对应于SEQ ID NO:1、3、36、40、42、46、49、52、56、59、62、64、66、72、74、76、77、80、81、84、86、87、105、107、88、109、130、134、136、137、140、143、146、149、152、155、157、159-163的氨基酸序列的CoV刺突(S)多肽中切割信号肽后，获得具有选自SEQ ID NO:2、4、38、41、44、48、51、54、58、61、63、65、67、73、75、78、79、82、83、85、106、108、89和110、112-115、132、133、114、138、141、144、147、151、153、156和158、164-168的氨基酸序列的成熟多肽，并且将其用于产生CoV S纳米颗粒疫苗或CoV S纳米颗粒。

有利地，相对于天然CoV刺突(S)蛋白，所公开的CoV S多肽可以具有增强的蛋白质表达和稳定性。

在实施方案中，本申请所述的CoV S多肽含有相对于天然冠状病毒S蛋白(SEQ IDNO:2)的进一步修饰。在实施方案中，本申请所述的冠状病毒S蛋白与天然冠状病毒S蛋白展现出至少80％、或至少90％、或至少95％、或至少97％、或至少99％同一性。本领域技术人员将使用已知技术来计算重组冠状病毒S蛋白与天然蛋白或本申请所述的任何CoV S多肽的同一性百分比。例如，可以使用可在线获得的工具CLUSTALW2或基本局部比对搜索工具(Basic Local Alignment Search Tool，BLAST)来计算同一性百分比。可以将以下默认参数用于CLUSTALW2配对比对：蛋白质权重矩阵＝Gonnet；空位开放＝10；空位延伸＝0.1。

在实施方案中，本申请所述的CoV S多肽与具有SEQ ID NO:87的氨基酸序列的CoVS多肽至少95％、至少96％、至少97％、至少98％、至少99％或至少99.5％相同。与具有SEQID NO:87的氨基酸序列的CoV S多肽的氨基酸序列相比，CoV S多肽可以具有多达约1、多达约2、多达约3、多达约4、多达约5、多达约10、多达约15、多达约20、多达约25、多达约30、多达约35、多达约40、多达约45或多达约50个氨基酸的缺失、插入或突变。与具有SEQ ID NO:87的氨基酸序列的CoV S多肽相比，CoV S多肽可以具有在约1与约5个之间的氨基酸、在约3与约10个之间的氨基酸、在约5与10个之间的氨基酸、在约8与12个之间的氨基酸、在约10与15个之间的氨基酸、在约12与17个之间的氨基酸、在约15与20个之间的氨基酸、在约18与23个之间的氨基酸、在约20与25个之间的氨基酸、在约22与约27个之间的氨基酸、在约25与30个之间的氨基酸、在约30与35个之间的氨基酸、在约35与40个之间的氨基酸、在约40与45个之间的氨基酸或在约45与50个之间的氨基酸的缺失、插入或突变。在实施方案中，与冠状病毒S蛋白(SEQ ID NO:87)相比，本申请所述的CoV S多肽包含约1、约2、约3、约4、约5、约6、约7、约8、约9、约10、约11、约12、约13、约14、约15、约16、约17、约18、约19、约20、约21、约22、约23、约24或约25个取代。

在实施方案中，本申请所述的CoV S多肽与具有选自SEQ ID NO:2、4、38、41、44、48、51、54、58、61、63、65、67、73、75、78、79、82、83、85、106、108、89和110、112-115、132、133、114、138、141、144、147、151、153、156和158、164-168中任一个的氨基酸序列的CoV S多肽至少95％、至少96％、至少97％、至少98％、至少99％或至少99.5％相同。与具有选自SEQ IDNO:2、4、38、41、44、48、51、54、58、61、63、65、67、73、75、78、79、82、83、85、106、108、89和110、112-115、132、133、114、138、141、144、147、151、153、156和158、164-168中任一个的氨基酸序列的CoV S多肽的氨基酸序列相比，CoV S多肽可以具有多达约1、多达约2、多达约3、多达约4、多达约5、多达约10、多达约15、多达约20、多达约25、多达约30、多达约35、多达约40、多达约45或多达约50个氨基酸的缺失、插入或突变。与具有选自SEQ ID NO:2、4、38、41、44、48、51、54、58、61、63、65、67、73、75、78、79、82、83、85、106、108、89和110、112-115、132、133、114、138、141、144、147、151、153、156 1和58、164-168中任一个的氨基酸序列的CoV S多肽相比，CoV S多肽可以具有在约1与约5个之间的氨基酸、在约3与约10个之间的氨基酸、在约5与10个之间的氨基酸、在约8与12个之间的氨基酸、在约10与15个之间的氨基酸、在约12与17个之间的氨基酸、在约15与20个之间的氨基酸、在约18与23个之间的氨基酸、在约20与25个之间的氨基酸、在约22与约27个之间的氨基酸、在约25与30个之间的氨基酸、在约30与35个之间的氨基酸、在约35与40个之间的氨基酸、在约40与45个之间的氨基酸或在约45与50个之间的氨基酸的缺失、插入或突变。在实施方案中，与冠状病毒S蛋白(SEQ ID NO:87)相比，本申请所述的CoV S多肽包含约1、约2、约3、约4、约5、约6、约7、约8、约9、约10、约11、约12、约13、约14、约15、约16、约17、约18、约19、约20、约21、约22、约23、约24或约25个取代。

在实施方案中，所述冠状病毒S多肽在N末端、C末端或N末端和C末端两者处延伸。在各个方面，所述延伸物是对诸如纯化或检测等功能有用的标签。在各个方面，所述标签含有表位。例如，所述标签可以是聚谷氨酸标签、FLAG标签、HA标签、聚His标签(具有约5-10个组氨酸)(SEQ ID NO:101)、六组氨酸标签(SEQ ID NO:100)、8X-His标签(具有八个组氨酸)(SEQ ID NO:102)、Myc标签、谷胱甘肽-S-转移酶标签、绿色荧光蛋白标签、麦芽糖结合蛋白标签、硫氧还蛋白标签或Fc标签。在其他方面，所述延伸物可以是与蛋白质融合以增强表达的N末端信号肽。尽管此类信号肽通常在细胞中表达期间被切割，但是一些纳米颗粒可包含具有完整信号肽的抗原。因此，当纳米颗粒包含抗原时，所述抗原可以含有延伸物，因此当掺入纳米颗粒中时可以是融合蛋白。出于计算与序列的同一性的目的，不包括延伸物。在实施方案中，所述标签是蛋白酶切割位点。蛋白酶切割位点的非限制性例子包括HRV3C蛋白酶切割位点、胰凝乳蛋白酶、胰蛋白酶、弹性蛋白酶、内肽酶、半胱天冬酶1、半胱天冬酶2、半胱天冬酶3、半胱天冬酶4、半胱天冬酶5、半胱天冬酶6、半胱天冬酶7、半胱天冬酶8、半胱天冬酶9、半胱天冬酶10、肠激酶、因子Xa、颗粒酶B、TEV蛋白酶和凝血酶。在实施方案中，所述蛋白酶切割位点是HRV3C蛋白酶切割位点。在实施方案中，所述蛋白酶切割位点包含SEQ IDNO:98的氨基酸序列。

在实施方案中，所述CoV S糖蛋白包含融合蛋白。在实施方案中，所述CoV S糖蛋白包含N末端融合蛋白。在实施方案中，所述CoV S糖蛋白包含C末端融合蛋白。在实施方案中，所述融合蛋白包括可用于蛋白质表达、纯化或检测的标签。在实施方案中，所述标签是聚His标签(具有约5-10个组氨酸)、Myc标签、谷胱甘肽-S-转移酶标签、绿色荧光蛋白标签、麦芽糖结合蛋白标签、硫氧还蛋白标签、Strep标签、Twin-Strep标签或Fc标签。在实施方案中，所述标签是Fc标签。在实施方案中，所述Fc标签是单体、二聚体或三聚体的。在实施方案中，所述标签是六组氨酸标签，例如含有六个组氨酸的聚His标签(SEQ ID NO:100)。在实施方案中，所述标签是具有SEQ ID NO:99的氨基酸序列的Twin-Strep标签。

在实施方案中，所述CoV S多肽是包含另一种冠状病毒蛋白的融合蛋白。在实施方案中，另一种冠状病毒蛋白来自相同的冠状病毒。在实施方案中，另一种冠状病毒蛋白来自不同的冠状病毒。

在各个方面，所述CoV S蛋白可以被截短。例如，N末端可被截短约10个氨基酸、约30个氨基酸、约50个氨基酸、约75个氨基酸、约100个氨基酸或约200个氨基酸。代替N末端或除了N末端之外，C末端可以是截短的。例如，C末端可以被截短约10个氨基酸、约30个氨基酸、约50个氨基酸、约75个氨基酸、约100个氨基酸或约200个氨基酸。出于计算与具有截短的蛋白质的同一性的目的，在蛋白质的剩余部分上测量同一性。

含有CoV刺突(S)多肽的纳米颗粒

在实施方案中，使用成熟CoV S多肽抗原来产生包含冠状病毒S纳米颗粒的疫苗。在实施方案中，本申请公开文本的纳米颗粒包含本申请所述的CoV S多肽。在实施方案中，本申请公开文本的纳米颗粒包含与洗涤剂核心缔合的CoV S多肽。洗涤剂的存在通过形成组织并呈递抗原的核心来促进纳米颗粒的形成。在实施方案中，所述纳米颗粒以含有组装成多寡聚糖蛋白-洗涤剂(例如PS80)纳米颗粒的CoV S多肽，其中头部区域向外突出并且疏水区域和PS80洗涤剂形成被糖蛋白包围的中央核心。在实施方案中，所述CoV S多肽固有地含有或适于含有跨膜结构域以促进将所述蛋白质缔合到洗涤剂核心中。在实施方案中，所述CoV S多肽含有头部结构域。图10显示了本申请公开文本的CoV S多肽的示例性结构。主要地，CoV S多肽三聚体的跨膜结构域与洗涤剂缔合；然而，多肽的其他部分也可以相互作用。有利地，所述纳米颗粒具有改善的对环境胁迫的抗性，使得由于在洗涤剂周围组织蛋白质的多个拷贝，它们提供增强的稳定性和/或改善的对免疫系统的呈递。

在实施方案中，所述洗涤剂核心是非离子型洗涤剂核心。在实施方案中，所述CoVS多肽与非离子型洗涤剂核心缔合。在实施方案中，所述洗涤剂选自聚山梨醇酯20(PS20)、聚山梨醇酯40(PS40)、聚山梨醇酯60(PS60)、聚山梨醇酯65(PS65)和聚山梨醇酯80(PS80)。

在实施方案中，所述洗涤剂是PS80。

在实施方案中，所述CoV S多肽形成三聚体。在实施方案中，所述CoV S多肽纳米颗粒由围绕非离子型洗涤剂核心的多个多肽三聚体构成。在实施方案中，所述纳米颗粒含有至少约1个或多个三聚体。在实施方案中，所述纳米颗粒含有刺突蛋白的至少约5个三聚体至约30个三聚体。在实施方案中，每个纳米颗粒可以含有1、2、3、4、5、6、7、8、9、10、11、12、或15、20、25或30个三聚体，包括在中间的所有值和范围。本申请所公开的组合物可以含有具有不同数量的三聚体的纳米颗粒。例如，组合物可以含有三聚体的数量范围从2-9的纳米颗粒；在实施方案中，组合物中的纳米颗粒可以含有2-6个三聚体。在实施方案中，所述组合物含有纳米颗粒的异质群体，每个纳米颗粒具有2至6个三聚体，或者每个纳米颗粒具有2至9个三聚体。在实施方案中，所述组合物可以含有纳米颗粒的基本上均质的群体。例如，群体可包含约95％的具有5个三聚体的纳米颗粒。

本申请所公开的纳米颗粒的颗粒尺寸具有一定范围。在实施方案中，本申请所公开的纳米颗粒的颗粒尺寸的Z-ave尺寸范围从约20nm至约60nm、约20nm至约50nm、约20nm至约45nm、约20nm至约35nm、约20nm至约30nm、约25nm至约35nm或约25nm至约45nm。除非另外说明，否则使用Zetasizer NanoZS(Malvern，英国)通过动态光散射(DLS)来测量颗粒尺寸(Z-ave)。

在实施方案中，与包含野生型CoV S多肽的纳米颗粒相比，包含本申请所公开的CoV S多肽的纳米颗粒具有减小的颗粒尺寸。在实施方案中，所述CoV S多肽的颗粒尺寸小至少约40％，例如颗粒尺寸小至少约40％、至少约45％、至少约50％、至少约55％、至少约60％、至少约65％、至少约70％、至少约75％、至少约80％或至少约85％。

包含本申请所公开的CoV S多肽的纳米颗粒在尺寸、形状和质量方面比包含野生型CoV S多肽的纳米颗粒更均匀。多分散性指数(PDI)是异质性的度量，除非另外说明，否则使用Malvern Setasizer通过动态光散射来测量。在实施方案中，本申请测量的颗粒的PDI从约0.2至约0.45，例如约0.2、约0.25、约0.29、约0.3、约0.35、约0.40或约0.45。在实施方案中，本申请测量的纳米颗粒的PDI比包含野生型CoV S多肽的纳米颗粒的PDI小至少约25％，例如小至少约25％、至少约30％、至少约35％、至少约40％、至少约45％、至少约50％、至少约55％或至少约60％。

与野生型CoV S多肽或其纳米颗粒相比，所述CoV S多肽和包含所述多肽的纳米颗粒具有改善的热稳定性。除非另外说明，否则使用差示扫描量热法(DSC)来测量CoV S多肽的热稳定性。转变焓(ΔHcal)是使CoV S多肽展开所需的能量。在实施方案中，与野生型CoVS多肽相比，所述CoV S多肽具有增加的ΔHcal。在实施方案中，CoV S多肽的ΔHcal是野生型CoV S多肽的ΔHcal的约2倍、约3倍、约4倍、约5倍、约6倍、约7倍、约8倍、约9倍或约10倍。

在本申请所公开的疫苗组合物中可以包括若干种纳米颗粒类型。在各个方面，纳米颗粒类型呈各向异性棒的形式，其可以是二聚体或单体。在其他方面，纳米颗粒类型是球形寡聚物。在又其他方面，所述纳米颗粒可以被描述为中间体纳米颗粒，其具有介于前两种类型之间的沉降特性。可以通过控制产生过程期间的洗涤剂和蛋白质浓度来调节纳米颗粒类型的形成。可以通过测量沉降系数来确定纳米颗粒类型。

含有CoV S多肽抗原的纳米颗粒的产生

本申请公开文本的纳米颗粒是非天然存在的产物，其组分在自然界中不会一起出现。通常，本申请所公开的方法使用洗涤剂交换方法，其中使用第一洗涤剂来分离蛋白质，然后将该第一洗涤剂交换成第二洗涤剂以形成纳米颗粒。

所述纳米颗粒中包含的抗原通常是通过在宿主细胞中重组表达而产生的。可以使用标准重组技术。在实施方案中，使用杆状病毒系统在昆虫宿主细胞中表达所述CoV S多肽。在实施方案中，所述杆状病毒是组织蛋白酶-L敲除的杆状病毒、几丁质酶敲除的杆状病毒。任选地，所述杆状病毒是组织蛋白酶-L和几丁质酶两者的双重敲除。在昆虫细胞表达系统中可以获得高水平的表达。昆虫细胞的非限制性例子是草地贪夜蛾(Spodopterafrugiperda)(Sf)细胞(例如Sf9、Sf21)、粉纹夜蛾(Trichoplusiani)细胞(例如High Five细胞)和果蝇(Drosophila)S2细胞。在实施方案中，在任何合适的宿主细胞中产生本申请所述的CoV S多肽。在实施方案中，所述宿主细胞是昆虫细胞。在实施方案中，所述昆虫细胞是Sf9细胞。

典型的转染和细胞生长方法可以用于培养所述细胞。可以根据本领域熟知的方法将载体(例如，包含编码融合蛋白的多核苷酸的载体)转染到宿主细胞中。例如，可以通过磷酸钙共沉淀、电穿孔、显微注射、脂质转染和采用多胺转染试剂的转染来实现将核酸引入真核细胞中。在一个实施方案中，所述载体是重组杆状病毒。

使宿主细胞生长的方法包括但不限于分批、补料分批、连续和灌注细胞培养技术。细胞培养意指细胞在生物反应器(发酵室)中的生长和繁殖，在所述生物反应器中细胞繁殖并表达蛋白质(例如重组蛋白)以用于纯化和分离。通常，细胞培养是在生物反应器中在无菌、受控温度和大气条件下进行。生物反应器是可以监测环境条件诸如温度、大气、搅拌和/或pH的用于培养细胞的室。在一个实施方案中，所述生物反应器是不锈钢室。在另一个实施方案中，所述生物反应器是预灭菌的塑料袋(例如Wave Biotech，新泽西州布里奇沃特)。在其他实施方案中，所述预灭菌的塑料袋是约50L至3500L的袋。

含有CoV刺突(S)蛋白抗原的纳米颗粒的提取和纯化

在宿主细胞生长后，可以使用洗涤剂和纯化方案从宿主细胞中收获所述蛋白质。一旦宿主细胞已生长48至96小时，便从培养基中分离细胞，并且添加含洗涤剂的溶液以溶解细胞膜，从而使蛋白质释放到洗涤剂提取物中。Triton X-100和壬基酚乙氧基化物(也称为NP-9)各自是用于提取的优选洗涤剂。可以将洗涤剂添加至约0.1％至约1.0％的最终浓度。例如，浓度可以为约0.1％、约0.2％、约0.3％、约0.5％、约0.7％、约0.8％或约1.0％。范围可以是约0.1％至约0.3％。在各个方面，浓度为约0.5％。

在其他方面，不同的第一洗涤剂可以用于从宿主细胞中分离所述蛋白质。例如，所述第一洗涤剂可以是双(聚乙二醇双[咪唑基羰基])、壬苯醇醚-9、双(聚乙二醇双[咪唑基羰基])、聚乙二醇十二烷基醚35、聚乙二醇(3)十六烷基醚56、醇乙氧基化物72、聚乙二醇2硬脂基醚76、聚乙二醇单油烯基醚92V、聚氧乙烯(10)油烯基醚97、聚乙二醇十六烷基醚58P、EL聚乙二醇甘油蓖麻酸酯、十乙二醇单十二烷基醚、N-癸酰基-N-甲基葡糖胺、正癸基α-D吡喃葡萄糖苷、癸基β-D-麦芽吡喃糖苷、正十二烷酰基-N-甲基葡糖酰胺、正十二烷基α-D-麦芽糖苷、正十二烷基β-D-麦芽糖苷、正十二烷基β-D-麦芽糖苷、七乙二醇单癸基醚、七乙二醇单十二烷基醚、七乙二醇单十四烷基醚、正十六烷基β-D-麦芽糖苷、六乙二醇单十二烷基醚、六乙二醇单十六烷基醚、六乙二醇单十八烷基醚、六乙二醇单十四烷基醚、Igepal CA-630、Igepal CA-630、甲基-6-0-(N-庚基氨基甲酰基)-α-D-吡喃葡萄糖苷、九乙二醇单十二烷基醚、N-壬酰基-N-甲基葡糖胺、N-壬酰基N-甲基葡糖胺、八乙二醇单癸基醚、八乙二醇单十二烷基醚、八乙二醇单十六烷基醚、八乙二醇单十八烷基醚、八乙二醇单十四烷基醚、辛基-β-D吡喃葡萄糖苷、五乙二醇单癸基醚、五乙二醇单十二烷基醚、五乙二醇单十六烷基醚、五乙二醇单己基醚、五乙二醇单十八烷基醚、五乙二醇单辛基醚、聚乙二醇二缩水甘油醚、聚乙二醇醚W-1、聚氧乙烯10十三烷基醚、聚氧乙烯100硬脂酸酯、聚氧乙烯20异十六烷基醚、聚氧乙烯20油烯基醚、聚氧乙烯40硬脂酸酯、聚氧乙烯50硬脂酸酯、聚氧乙烯8硬脂酸酯、聚氧乙烯双(咪唑基羰基)、聚氧乙烯25丙二醇硬脂酸酯、皂草苷(Saponin from Quillaja bark)、20山梨聚糖月桂酸酯、40山梨聚糖单棕榈酸酯、60山梨聚糖硬脂酸酯、65山梨聚糖三硬脂酸酯、80山梨聚糖单油酸酯、85山梨聚糖三油酸酯、二级醇乙氧基化物15-S-12型、二级醇乙氧基化物15-S-30型、二级醇乙氧基化物15-S-5型、二级醇乙氧基化物15-S-7型、二级醇乙氧基化物15-S-9型、壬基酚乙氧基化物NP-10型、壬基酚乙氧基化物NP-4型、壬基酚乙氧基化物NP-40型、壬基酚乙氧基化物NP-7型、壬基酚乙氧基化物NP-9型、支链二级醇乙氧基化物TMN-10型、支链二级醇乙氧基化物TMN-6型、TRITON^TM X-100聚乙二醇三级辛基苯基醚或其组合。

然后可以使用离心从细胞碎片中分离纳米颗粒。在实施方案中，可以使用诸如采用氯化铯、蔗糖和碘克沙醇的梯度离心。其他技术可以用作替代或补充，例如标准纯化技术，包括例如离子交换层析、亲和层析和凝胶过滤层析。

例如，第一柱可以是离子交换层析树脂，诸如EMD基于甲基丙烯酸酯的聚合物珠TMAE(EMD Millipore)；第二柱可以是小扁豆(Lens culinaris)凝集素亲和树脂；并且第三柱可以是阳离子交换柱，诸如EMD基于甲基丙烯酸酯的聚合物珠SO3(EMD Millipore)树脂。在其他方面，阳离子交换柱可以是MMC柱或Nuvia C Prime柱(Bio-Rad Laboratories,Inc)。优选地，本申请所公开的方法不使用洗涤剂提取柱；例如疏水相互作用柱。这样的柱通常在纯化期间用于去除洗涤剂，但可能对此处公开的方法产生负面影响。

含有CoV S多肽抗原的纳米颗粒的洗涤剂交换

为了形成纳米颗粒，用于从宿主细胞中提取蛋白质的第一洗涤剂基本上被第二洗涤剂替代以得到纳米颗粒结构。NP-9是优选的提取洗涤剂。通常，当通过HPLC测量时，所述纳米颗粒不含有可检测的NP-9。所述第二洗涤剂通常选自PS20、PS40、PS60、PS65和PS80。优选地，所述第二洗涤剂是PS80。

在特定方面，使用亲和层析以结合糖蛋白(经由其碳水化合物部分)来进行洗涤剂交换。例如，亲和层析可以使用豆类凝集素柱。豆类凝集素是最初在植物中鉴定出的蛋白质，并且发现其与碳水化合物残基特异性地且可逆地相互作用。参见例如，Sharon和Lis,“Legume lectins--a large family of homologous proteins,”FASEB J.1990年11月；4(14):3198-208；Liener,“The Lectins:Properties,Functions,and Applications inBiology and Medicine,”Elsevier,2012。合适的凝集素包括伴刀豆球蛋白A(con A)、豌豆凝集素、红豆草凝集素和小扁豆凝集素。小扁豆凝集素由于其结合特性是用于洗涤剂交换的优选柱。凝集素柱可商购获得；例如，Capto小扁豆凝集素可从GE Healthcare获得。在某些方面，小扁豆凝集素柱可以使用重组凝集素。在分子水平上，认为碳水化合物部分与小扁豆凝集素结合，释放蛋白质的氨基酸以在洗涤剂周围聚结，导致形成洗涤剂核心，从而提供具有多个抗原拷贝(例如，糖蛋白寡聚体)的纳米颗粒，所述糖蛋白寡聚体可以是锚定在洗涤剂中的二聚体、三聚体或四聚体。在实施方案中，所述CoV S多肽形成三聚体。在实施方案中，所述CoV S多肽三聚体锚定在洗涤剂中。在实施方案中，每个CoV S多肽纳米颗粒含有至少一个与非离子型核心缔合的三聚体。

当在洗涤剂交换期间与蛋白质一起孵育形成纳米颗粒时，洗涤剂在早期纯化步骤期间能以多达约0.1％(w/v)存在，并且使此量降低以获得具有最佳稳定性的最终纳米颗粒。例如，所述非离子型洗涤剂(例如，PS80)可以是约0.005％(v/v)至约0.1％(v/v)，例如约0.005％(v/v)、约0.006％(v/v)、约0.007％(v/v)、约0.008％(v/v)、约0.009％(v/v)、约0.01％(v/v)、约0.015％(v/v)、约0.02％(v/v)、约0.025％(v/v)、约0.03％(v/v)、约0.035％(v/v)、约0.04％(v/v)、约0.045％(v/v)、约0.05％(v/v)、约0.055％(v/v)、约0.06％(v/v)、约0.065％(v/v)、约0.07％(v/v)、约0.075％(v/v)、约0.08％(v/v)、约0.085％(v/v)、约0.09％(v/v)、约0.095％(v/v)或约0.1％(v/v)PS80。在实施方案中，所述纳米颗粒含有约0.03％至约0.05％PS80。在实施方案中，所述纳米颗粒含有约0.01％(v/v)PS80。

在实施方案中，对经纯化的CoV S多肽进行透析。在实施方案中，在纯化后进行透析。在实施方案中，在包含磷酸钠、NaCl和PS80的溶液中对所述CoV S多肽进行透析。在实施方案中，包含磷酸钠的透析溶液含有在约5mM与约100mM之间的磷酸钠，例如约5mM、约10mM、约15mM、约20mM、约25mM、约30mM、约35mM、约40mM、约45mM、约50mM、约55mM、约60mM、约65mM、约70mM、约75mM、约80mM、约85mM、约90mM、约95mM或约100mM磷酸钠。在实施方案中，包含磷酸钠的溶液的pH为约6.5、约6.6、约6.7、约6.8、约6.9、约7.0、约7.1、约7.2、约7.3、约7.4或约7.5。在实施方案中，包含氯化钠的透析溶液包含约50mM NaCl至约500mM NaCl，例如约50mM、约60mM、约70mM、约80mM、约90mM、约100mM、约110mM、约120mM、约130mM、约140mM、约150mM、约160mM、约170mM、约180mM、约190mM、约200mM、约210mM、约220mM、约230mM、约240mM、约250mM、约260mM、约270mM、约280mM、约290mM、约300mM、约310mM、约320mM、约330mM、约340mM、约350mM、约360mM、约370mM、约380mM、约390mM、约400mM、约410mM、约420mM、约430mM、约440mM、约450mM、约460mM、约470mM、约480mM、约490mM或约500mM NaCl。在实施方案中，包含PS80的透析溶液包含约0.005％(v/v)、约0.006％(v/v)、约0.007％(v/v)、约0.008％(v/v)、约0.009％(v/v)、约0.01％(v/v)、约0.015％(v/v)、约0.02％(v/v)、约0.025％(v/v)、约0.03％(v/v)、约0.035％(v/v)、约0.04％(v/v)、约0.045％(v/v)、约0.05％(v/v)、约0.055％(v/v)、约0.06％(v/v)、约0.065％(v/v)、约0.07％(v/v)、约0.075％(v/v)、约0.08％(v/v)、约0.085％(v/v)、约0.09％(v/v)、约0.095％(v/v)或约0.1％(v/v)PS80。在实施方案中，所述透析溶液包含约25mM磷酸钠(pH 7.2)、约300mM NaCl和约0.01％(v/v)PS80。

可以使用如下蛋白质进行洗涤剂交换，将所述蛋白质如上文所讨论地纯化，并且纯化、冷冻储存，然后解冻以进行洗涤剂交换。

本申请所公开的组合物的稳定性可以按多种方式测量。在一种方法中，可以制备肽图谱，以在设计成通过模拟苛刻的储存条件来胁迫纳米颗粒的各种处理后确定抗原蛋白的完整性。因此，稳定性的度量是与对照样品相比胁迫样品中抗原肽的相对丰度。例如，可以通过将含有CoV S多肽的纳米颗粒暴露于各种pH、蛋白酶、盐、氧化剂(包括但不限于过氧化氢)、各种温度、冷冻/解冻循环和搅拌来评价所述纳米颗粒的稳定性。图12A-图12B显示BV2373(SEQ ID NO:87)和BV2365(SEQ ID NO:4)在多种胁迫条件下保持与hACE2结合。认为糖蛋白锚定在洗涤剂核心中的位置通过减少不希望的相互作用而提供增强的稳定性。例如，可以通过屏蔽作用来实现针对基于蛋白酶的降解的改善的保护，由此以本申请所公开的摩尔比将糖蛋白锚定在核心中导致了空间位阻，从而阻断蛋白酶进入。也可以通过监测完整蛋白质来测量稳定性。图33和图34比较了含有分别具有SEQ ID NO:109和87的氨基酸序列的CoV多肽的纳米颗粒。图34表明，具有SEQ ID NO:87的氨基酸序列的CoV S多肽在纯化期间显示出特别好的稳定性。图34的多肽包含具有QQAQ(SEQ ID NO:7)的氨基酸序列的弗林蛋白酶切割位点。

含有CoV S多肽抗原的疫苗组合物

本申请公开文本提供了例如在纳米颗粒中包含CoV S多肽的疫苗组合物。在各个方面，疫苗组合物可以包含具有抗原的纳米颗粒，所述抗原来自同一病毒物种的超过一种病毒株。在另一个实施方案中，本申请公开文本提供了包含一个或多个容器的药物包装或试剂盒，所述一个或多个容器装有疫苗组合物的一种或多种组分。

本申请所公开的组合物可以预防性地或治疗性地使用，但是通常将是预防性的。因此，本申请公开文本包括用于治疗或预防感染的方法。所述方法涉及向受试者施用治疗量或预防量的本申请公开文本的免疫原性组合物。优选地，药物组合物是提供保护作用的疫苗组合物。在其他方面，所述保护作用可以包括在一定百分比的暴露人群中改善与感染相关的症状。例如，与未经治疗的受试者相比，所述组合物可以预防或减轻选自以下的一种或多种病毒疾病症状：发热、疲劳、肌肉疼痛、头痛、喉咙痛、呕吐、腹泻、皮疹、肾和肝功能受损的症状、内出血和外出血。

可以在存在各种赋形剂、缓冲剂等的情况下将所述纳米颗粒配制用于作为疫苗施用。例如，疫苗组合物可以含有磷酸钠、氯化钠和/或组氨酸。磷酸钠可以以约10mM至约50mM、约15mM至约25mM或约25mM存在；在特定情况下，存在约22mM磷酸钠。组氨酸可以按约0.1％(w/v)、约0.5％(w/v)、约0.7％(w/v)、约1％(w/v)、约1.5％(w/v)、约2％(w/v)、或约2.5％(w/v)存在。氯化钠(当存在时)可以为约150mM。在某些组合物中，氯化钠可以按较高浓度(例如从约200mM至约500mM)存在。在实施方案中，氯化钠以高浓度(包括但不限于约200mM、约250mM、约300mM、约350mM、约400mM、约450mM或约500mM)存在。

在实施方案中，本申请所述的纳米颗粒在某些pH水平下具有改善的稳定性。在实施方案中，所述纳米颗粒在微酸性pH水平下是稳定的。例如，所述纳米颗粒在微酸性pH(例如从pH 5.8至pH 7.0)下是稳定的。在实施方案中，所述纳米颗粒和含有纳米颗粒的组合物在范围从约pH 5.8至约pH 7.0的pH(包括约pH 5.9至约pH 6.8、约pH 6.0至约pH 6.5、约pH6.1至约pH 6.4、约pH 6.1至约pH 6.3或约pH 6.2)下可以是稳定的。在实施方案中，本申请所述的纳米颗粒和组合物在中性pH(包括从约pH 7.0至约pH 7.4)下是稳定的。在实施方案中，本申请所述的纳米颗粒和组合物在微碱性pH(例如从约pH 7.0至约pH 8.5、从约pH 7.0至约pH 8.0或从约pH 7.0至约pH 7.5，包括在中间的所有值和范围)下是稳定的。

佐剂

在某些实施方案中，本申请所公开的组合物可以与一种或多种佐剂组合以增强免疫反应。在其他实施方案中，所述组合物是在没有佐剂的情况下制备的，因此可用来作为无佐剂组合物施用。有利地，当作为单剂量施用时，本申请所公开的无佐剂组合物可以提供保护性免疫反应。诱导稳健的免疫反应的无明矾组合物尤其可用于约60岁及以上的成人。

基于铝的佐剂

在实施方案中，所述佐剂可以是明矾(例如AlPO₄或Al(OH)₃)。通常，纳米颗粒基本上与明矾结合。例如，纳米颗粒可以是至少80％结合、至少85％结合、至少90％结合或至少95％结合至明矾。通常，在组合物中，纳米颗粒是92％至97％结合至明矾。每个剂量存在的明矾的量通常在约400μg至约1250μg之间的范围内。例如，明矾可以按每个剂量约300μg至约900μg、约400μg至约800μg、约500μg至约700μg、约400μg至约600μg、或约400μg至约500μg的量存在。通常，对于120μg蛋白纳米颗粒的剂量，明矾以约400μg存在。

皂苷佐剂

含有皂苷的佐剂也可以与本申请所公开的免疫原组合。皂苷是源自皂树树皮的糖苷。通常，使用多步纯化过程制备皂苷，从而产生多种级分。如本申请所用，术语“来自皂树的皂苷级分”一般用于描述皂树的半纯化或确定的皂苷级分或其基本上纯的级分。

皂苷级分

用于产生皂苷级分的若干种方法是合适的。级分A、B和C描述于美国专利号6,352,697中并且可以如下制备。通过层析法分离来自Quil A(一种粗制水性皂树提取物)的亲脂级分，并将其用水中的70％乙腈洗脱以回收亲脂级分。然后通过半制备型HPLC分离这种亲脂级分，使用在酸性水中的从25％至60％乙腈的梯度进行洗脱。在本申请中称为“级分A”或“QH-A”的级分是或对应于在大约39％乙腈下洗脱的级分。在本申请中称为“级分B”或“QH-B”的级分是或对应于在大约47％乙腈下洗脱的级分。在本申请中称为“级分C”或“QH-C”的级分是或对应于在大约49％乙腈下洗脱的级分。关于级分纯化的另外信息可以在美国专利号5,057,540中找到。当如本申请所述制备时，皂树的级分A、B和C各自代表具有可定义特性的化学上紧密相关分子的组或家族。获得所述级分的层析条件使得就洗脱曲线和生物活性而言批次间重现性是高度一致的。

已经描述了其他皂苷级分。级分B3、B4和B4b描述于EP 0436620中。级分QA1-QA22是描述的EP03632279 B2，Q-VAC(Nor-Feed,ASDenmark)，皂树Spikoside(lsconova AB,Ultunaallén 2B,756 51乌普萨拉,瑞典)。可以使用EP 0 3632 279B2中的级分QA-1、QA-2、QA-3、QA-4、QA-5、QA-6、QA-7、QA-8、QA-9、QA-10、QA-11、QA-12、QA-13、QA-14、QA-15、QA-16、QA-17、QA-18、QA-19、QA-20、QA-21、和QA-22，尤其是QA-7、QA-17、QA-18、和QA-21。如EP 03632 279B2中(尤其是在第6页以及在第8和9页中的实例1中)所述获得所述级分。

本申请所述并用于形成佐剂的皂苷级分通常是基本上纯的级分；也就是说，所述级分基本上不存在来自其他材料的污染。在特定方面，基本上纯的皂苷级分可以含有按重量计至多40％、按重量计至多30％、按重量计至多25％、按重量计至多20％、按重量计至多15％、按重量计至多10％、按重量计至多7％、按重量计至多5％、按重量计至多2％、按重量计至多1％、按重量计至多0.5％或按重量计至多0.1％的其他化合物，诸如其他皂苷或其他佐剂材料。

ISCOM结构

皂苷级分能以称为ISCOM(免疫刺激复合物)的笼状颗粒的形式施用。ISCOM可以如EP0109942B1、EP0242380B1和EP0180546 B1中所述制备。在特定实施方案中，可以使用转运和/或乘客抗原，如EP 9600647-3(PCT/SE97/00289)中所述。

基质佐剂

在实施方案中，所述ISCOM是ISCOM基质复合物。所述ISCOM基质复合物包含至少一种皂苷级分和脂质。所述脂质至少是固醇，诸如胆固醇。在特定方面，所述ISCOM基质复合物还含有磷脂。所述ISCOM基质复合物还可以含有一种或多种其他免疫调节(佐剂活性)物质，不一定是糖苷，并且可以如EP0436620B1中所述产生，将所述专利通过引用以其整体并入本申请。

在其他方面，所述ISCOM是ISCOM复合物。ISCOM复合物含有至少一种皂苷、至少一种脂质和至少一种抗原或表位。所述ISCOM复合物含有通过洗涤剂处理缔合的抗原，使得一部分抗原整合到颗粒中。相比之下，ISCOM基质被配制为与抗原的混合物，并且ISCOM基质颗粒与抗原之间的缔合通过静电和/或疏水相互作用来介导。

根据一个实施方案，整合到ISCOM基质复合物或ISCOM复合物中的皂苷级分或也整合到ISCOM或ISCOM基质复合物中或与其混合的至少一种另外的佐剂选自皂树的级分A、级分B或级分C、皂树的半纯化制剂、皂树的纯化制剂或任何经纯化的亚级分(例如，QA 1-21)。

在特定方面，每个ISCOM颗粒可以含有至少两种皂苷级分。可以使用不同皂苷级分的重量％的任何组合。可以使用任何两种级分的重量％的任何组合。例如，所述颗粒可以分别含有任何重量％的级分A和任何重量％的另一种皂苷级分，诸如粗皂苷级分或级分C。因此，在特定方面，每个ISCOM基质颗粒或每个ISCOM复合物颗粒可以含有按重量计0.1％至99.9％、按重量计5％至95％、按重量计10％至90％、按重量计15％至85％、按重量计20％至80％、按重量计25％至75％、按重量计30％至70％、按重量计35％至65％、按重量计40％至60％、按重量计45％至55％、按重量计40％至60％或按重量计50％的一种皂苷级分(例如级分A)以及在每种情况下其余多达100％的另一种皂苷(例如任何粗级分或任何其他级分，例如级分C)。所述重量被计算为皂苷级分的总重量。ISCOM基质复合物和ISCOM复合物佐剂的例子公开于美国公开的申请号2013/0129770中，将所述申请通过引用以其整体并入本申请。

在特定实施方案中，所述ISCOM基质或ISCOM复合物包含按重量计5％-99％的一种级分(例如级分A)和其余按重量计多达100％的另一种级分(例如粗皂苷级分或级分C)。所述重量被计算为皂苷级分的总重量。

在另一个实施方案中，所述ISCOM基质或ISCOM复合物包含按重量计40％至99％的一种级分(例如级分A)和按重量计1％至60％的另一种级分(例如粗皂苷级分或级分C)。所述重量被计算为皂苷级分的总重量。

在又另一个实施方案中，所述ISCOM基质或ISCOM复合物包含按重量计70％至95％的一种级分(例如，级分A)和按重量计30％至5％的另一种级分(例如，粗皂苷级分或级分C)。所述重量被计算为皂苷级分的总重量。在其他实施方案中，来自皂树的皂苷级分选自QA1-21中的任一种。

除了含有皂苷级分的混合物的颗粒之外，ISCOM基质颗粒和ISCOM复合物颗粒各自都可以仅使用一种皂苷级分形成。本申请所公开的组合物可以含有多个颗粒，其中每个颗粒仅含有一种皂苷级分。也就是说，某些组合物可以含有一种或多种不同类型的ISCOM-基质复合物颗粒和/或一种或多种不同类型的ISCOM复合物颗粒，其中每个单独的颗粒含有来自皂树的一种皂苷级分，其中一种复合物中的皂苷级分与另一种复合物颗粒中的皂苷级分不同。

在特定方面，一种类型的皂苷级分或粗皂苷级分可以整合到一种ISCOM基质复合物或颗粒中，并且另一种类型的基本上纯的皂苷级分或粗皂苷级分可以整合到另一种ISCOM基质复合物或颗粒中。组合物或疫苗可以包含至少两种类型的复合物或颗粒，每种类型具有一种类型的整合到物理上不同的颗粒中的皂苷。

在所述组合物中，可以使用ISCOM基质复合物颗粒和/或ISCOM复合物颗粒的混合物，其中将一种皂苷级分皂树和另一种皂苷级分皂树分开掺入不同的ISCOM基质复合物颗粒和/或ISCOM复合物颗粒中。

各自具有一种皂苷级分的ISCOM基质或ISCOM复合物颗粒可以按重量％的任何组合存在于组合物中。在特定方面，组合物可以含有按重量计0.1％至99.9％、按重量计5％至95％、按重量计10％至90％、按重量计15％至85％、按重量计20％至80％、按重量计25％至75％、按重量计30％至70％、按重量计35％至65％、按重量计40％至60％、按重量计45％至55％、按重量计40至60％或按重量计50％的含有第一皂苷级分的ISCOM基质或复合物，其余部分由含有不同皂苷级分的ISCOM基质或复合物构成。在各个方面，所述其余部分是一种或多种ISCOM基质或复合物，其中每个基质或复合物颗粒仅含有一种皂苷级分。在其他方面，所述ISCOM基质或复合物颗粒可以含有多于一种皂苷级分。

在特定组合物中，第一ISCOM基质或ISCOM复合物颗粒中唯一的皂苷级分是级分A，并且第二ISCOM基质或ISCOM复合物颗粒中唯一的皂苷级分是级分C。

优选的组合物包含含有级分A的第一ISCOM基质和含有级分C的第二ISCOM基质，其中级分A ISCOM基质占总皂苷佐剂重量的约70％，并且级分C ISCOM基质占总皂苷佐剂重量的约30％。在另一种优选的组合物中，级分A ISCOM基质占总皂苷佐剂重量的约85％，并且级分C ISCOM基质占总皂苷佐剂重量的约15％。因此，在某些组合物中，级分A ISCOM基质存在的范围为所述组合物中皂苷佐剂总重量的约70％至约85％，并且级分C ISCOM基质存在的范围为约15％至约30％。在实施方案中，分别地，级分A ISCOM基质占佐剂中级分A ISCOM基质和级分C ISCOM的重量之和的按重量计50％-96％，并且级分C ISCOM基质占剩余部分。在特别优选的组合物(在本申请中称为MATRIX-M^TM)中，级分A ISCOM基质以所述组合物中皂苷佐剂总重量的约85％存在，并且级分C ISCOM基质以约15％存在。MATRIX-M^TM可以可互换地称为Matrix-M1。

示例性QS-7和QS-21级分、其生产及其用途描述于美国专利号5,057,540、6,231,859、6,352,697、6,524,584、6,846,489、7,776,343和8,173,141中，将这些专利通过引用并入本申请。

在实施方案中，可以另外或替代地使用其他佐剂。包括在以下文献中所述的任何佐剂设想在本申请公开文本的范围内：Vogel等人,“A Compendium of Vaccine Adjuvantsand Excipients(第2版),”(出于所有目的通过引用以其整体并入本申请)。其他佐剂包括完全弗氏佐剂(免疫反应的非特异性刺激剂，含有杀死的结核分枝杆菌(Mycobacteriumtuberculosis))、不完全弗氏佐剂和氢氧化铝佐剂。其他佐剂包括GMCSP、BCG、MDP化合物(诸如thur-MDP和nor-MDP)、CGP(MTP-PE)、脂质A和单磷酰脂质A(MPL)、MF-59、RIBI(其含有从细菌中提取的三种组分)、MPL、海藻糖二霉菌酸酯(TDM)和在2％角鲨烯/聚山梨醇酯80乳剂中的细胞壁骨架(CWS)。在实施方案中，所述佐剂可以是少层状(paucilamellar)脂质囊泡；例如是范围从约100nm至约500nm的少层状非磷脂囊泡。它们包含醇乙氧基化物72、胆固醇、油酸和角鲨烯。已显示是有效的佐剂(参见美国专利号5,629,021、6,387,373和4,911,928)。

施用和剂量

在实施方案中，本申请公开文本提供了用于引发针对一种或多种冠状病毒的免疫反应的方法。在实施方案中，所述反应是针对SARS-CoV-2病毒、MERS和SARS中的一种或多种。在实施方案中，所述反应是针对异源SARS-CoV-2毒株。异源SARS-CoV-2毒株的非限制性例子包括Cal.20C SARS-CoV-2毒株、P.1SARS-CoV-2毒株、B.1.351SARS-CoV-2毒株和B.1.1.7SARS-CoV-2毒株。所述方法涉及向受试者施用免疫有效量的含有纳米颗粒或含有重组CoV刺突(S)多肽的组合物。有利地，本申请所公开的蛋白质诱导一种或多种特别有用的抗冠状病毒反应。

在实施方案中，将所述纳米颗粒或CoV S多肽与佐剂一起施用。在各个方面，在没有佐剂的情况下施用所述纳米颗粒或CoV S多肽。在各个方面，佐剂可以例如通过非共价相互作用与纳米颗粒结合。在其他方面，将佐剂与纳米颗粒共同施用，但是佐剂和纳米颗粒基本上不相互作用。

在实施方案中，所述纳米颗粒或CoV S多肽可以用于预防和/或治疗SARS-CoV-2感染、异源SARS-CoV-2毒株感染、SARS感染或MERS感染中的一种或多种。因此，本申请公开文本提供了用于引发针对SARS-CoV-2病毒、异源SARS-CoV-2病毒、MERS和SARS中的一种或多种的免疫反应的方法。所述方法涉及向受试者施用免疫有效量的含有纳米颗粒或CoV S多肽的组合物。有利地，本申请所公开的蛋白质诱导特别有用的抗冠状病毒反应。

在实施方案中，含有本申请所述的纳米颗粒或CoV S多肽的组合物在受试者中诱导针对SARS-CoV-2或异源SARS-CoV-2毒株的保护性反应，在最后一个剂量的纳米颗粒或CoV S多肽后所述保护性反应持续长达约3个月、长达约4个月、长达约5个月、长达约6个月、长达约7个月、长达约8个月、长达约9个月、长达约10个月、长达约11个月、长达约12个月、长达约13个月长达约14个月、长达约15个月、长达约16个月、长达约17个月、长达约18个月、长达约19个月、长达约20个月、长达约21个月、长达约22个月、长达约23个月、长达约24个月、长达约2.5年、长达约3年、长达约3.5年、长达约4年、长达约4.5年、长达约5年。在实施方案中，本申请所述的纳米颗粒或CoV S多肽在受试者中诱导保护性反应持续至少6个月。

在实施方案中，所述保护性反应是针对由SARS-CoV-2或异源SARS-CoV-2毒株引起的无症状感染。在实施方案中，所述保护性反应是针对由SARS-CoV-2或异源SARS-CoV-2毒株引起的症状性感染。

在实施方案中，含有本申请所述的纳米颗粒或CoV S多肽的组合物具有预防来自SARS-CoV-2病毒或异源SARS-CoV-2毒株(例如，B.1.1.7SARS-CoV-2毒株、B.1.351SARS-CoV-2毒株、P.1SARS-CoV-2毒株、B.1.617.2SARS-CoV-2毒株、B.1.525SARS-CoV-2毒株、B.1.526SARS-CoV-2毒株、B.1.617.1SARS-CoV-2毒株、C.37SARS-CoV-2毒株、B.1.621SARS-CoV-2毒株或Cal.20C SARS-CoV-2毒株)的冠状病毒19(COVID-19)的如下功效：约50％至约99％、约80％至约99％、约75％至约99％、约80％至约95％、约90％至约98％、约75％至约95％、约80％至约90％、约85％至约95％、约80％至约95％、至少约50％、至少约55％、至少约60％、至少约65％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％、至少约91％、至少约92％、至少约93％、至少约94％、至少约95％、至少约96％、至少约97％、至少约98％或至少约99％，所述功效在施用最后一个剂量的本申请所述的纳米颗粒或CoV S多肽后持续长达约1个月、长达约2个月、长达约2.5个月、长达约3个月、长达约3.5个月、长达约4个月、长达约4.5个月、长达约5个月、长达约5.5个月、长达约6个月、长达约6.5个月、长达约7个月、长达约7.5个月、长达约8个月、长达约8.5个月、长达约9个月、长达约9.5个月、长达约10个月、长达约10.5个月、长达约11个月、长达约11.5个月、长达约12个月、长达约12.5个月、长达约13个月、长达约13.5个月、长达约14个月、长达约14.5个月、长达约15个月、长达约15.5个月、长达约16个月、长达约16.5个月、长达约17个月、长达约17.5个月、长达约18个月、长达约18.5个月、长达约19个月、长达约19.5个月、长达约20个月、长达约20.5个月、长达约21个月、长达约21.5个月、长达约22个月、长达约22.5个月、长达约23个月、长达约23.5个月、长达约24个月、长达约2.1年、长达约2.2年、长达约2.3年、长达约2.4年、长达约2.5年、长达约2.6年、长达约2.7年、长达约2.8年、长达约2.9年、长达约3年或更长时间。在实施方案中，所述COVID-19是轻度COVID-19。在实施方案中，所述COVID-19是中度COVID-19。在实施方案中，所述COVID-19是重度COVID-19。在实施方案中，所述COVID-19是无症状COVID-19。

在实施方案中，含有本申请所述的纳米颗粒或CoV S多肽的组合物针对SARS-CoV-2病毒或异源SARS-CoV-2毒株具有至少82％的功效，所述功效在施用最后一个剂量的本申请所述的纳米颗粒或CoV S多肽后持续长达约7.5个月。在实施方案中，含有本申请所述的纳米颗粒或CoV S多肽的组合物针对SARS-CoV-2病毒或异源SARS-CoV-2毒株具有80％至约90％的功效，所述功效在施用最后一个剂量的本申请所述的纳米颗粒或CoV S多肽后持续长达约7.5个月。

在实施方案中，含有纳米颗粒或CoV S多肽的组合物针对无症状疾病具有至少75％的功效。在实施方案中，所述纳米颗粒或CoV S多肽针对症状性COVID-19具有如下功效：80％至90％、80％至99％、82％至99％、82％至95％、85％至95％、85％至99％、85至97％、至少80％、至少81％、至少82％、至少83％、至少84％、至少85％、至少86％、至少87％、至少88％、至少89％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、至少99％或100％，持续长达约1个月、长达约2个月、长达约2.5个月、长达约3个月、长达约3.5个月、长达约4个月、长达约4.5个月、长达约5个月、长达约5.5个月、长达约6个月、长达约6.5个月、长达约7个月、长达约7.5个月、长达约8个月、长达约8.5个月、长达约9个月、长达约9.5个月、长达约10个月、长达约10.5个月、长达约11个月、长达约11.5个月、长达约12个月、长达约12.5个月、长达约13个月、长达约13.5个月、长达约14个月、长达约14.5个月、长达约15个月、长达约15.5个月、长达约16个月、长达约16.5个月、长达约17个月、长达约17.5个月、长达约18个月、长达约18.5个月、长达约19个月、长达约19.5个月、长达约20个月、长达约20.5个月、长达约21个月、长达约21.5个月、长达约22个月、长达约22.5个月、长达约23个月、长达约23.5个月或长达约24个月或更长时间。

在实施方案中，含有所述纳米颗粒或CoV S多肽的组合物针对重度COVID-19具有如下功效：95％至97％、95％至99％、95％至98％、至少95％、至少96％、至少97％、至少98％、至少99％或100％，持续长达约1个月、长达约2个月、长达约2.5个月、长达约3个月、长达约3.5个月、长达约4个月、长达约4.5个月、长达约5个月、长达约5.5个月、长达约6个月、长达约6.5个月、长达约7个月、长达约7.5个月、长达约8个月、长达约8.5个月、长达约9个月、长达约9.5个月、长达约10个月、长达约10.5个月、长达约11个月、长达约11.5个月、长达约12个月、长达约12.5个月、长达约13个月、长达约13.5个月、长达约14个月、长达约14.5个月、长达约15个月、长达约15.5个月、长达约16个月、长达约16.5个月、长达约17个月、长达约17.5个月、长达约18个月、长达约18.5个月、长达约19个月、长达约19.5个月、长达约20个月、长达约20.5个月、长达约21个月、长达约21.5个月、长达约22个月、长达约22.5个月、长达约23个月、长达约23.5个月或长达约24个月或更长时间。

在实施方案中，含有所述纳米颗粒或CoV S多肽的组合物针对中度COVID-19具有如下功效：75％至95％、75％至90％、75％至85％、75％至98％、80％至98％、80％至95％、80％至90％、85％至98％、85％至95％、至少75％、至少76％、至少77％、至少78％、至少79％、至少80％、至少81％、至少82％、至少83％、至少84％、至少85％、至少86％、至少87％、至少88％、至少89％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、至少99％或100％，持续长达约1个月、长达约2个月、长达约2.5个月、长达约3个月、长达约3.5个月、长达约4个月、长达约4.5个月、长达约5个月、长达约5.5个月、长达约6个月、长达约6.5个月、长达约7个月、长达约7.5个月、长达约8个月、长达约8.5个月、长达约9个月、长达约9.5个月、长达约10个月、长达约10.5个月、长达约11个月、长达约11.5个月、长达约12个月、长达约12.5个月、长达约13个月、长达约13.5个月、长达约14个月、长达约14.5个月、长达约15个月、长达约15.5个月、长达约16个月、长达约16.5个月、长达约17个月、长达约17.5个月、长达约18个月、长达约18.5个月、长达约19个月、长达约19.5个月、长达约20个月、长达约20.5个月、长达约21个月、长达约21.5个月、长达约22个月、长达约22.5个月、长达约23个月、长达约23.5个月或长达约24个月或更长时间。

在实施方案中，含有所述纳米颗粒或CoV S多肽的组合物针对轻度COVID-19具有如下功效：40％至95％、40％至90％、40％至85％、40％至80％、40％至75％、40％至70％、40％至65％、40％至60％、50％至95％、50％至90％、50％至85％、50％至80％、50％至75％、50％至70％、50％至65％、50％至60％、至少40％、至少41％、至少42％、至少43％、至少44％、至少45％、至少46％、至少47％、至少48％、至少49％、至少50％、至少51％、至少52％、至少53％、至少54％、至少55％、至少56％、至少57％、至少58％、至少59％、至少60％、至少61％、至少62％、至少63％、至少64％、至少65％、至少66％、至少67％、至少68％、至少69％、至少70％、至少71％、至少72％、至少73％、至少74％、至少75％、至少76％、至少77％、至少78％、至少79％、至少80％、至少81％、至少82％、至少83％、至少84％、至少85％、至少86％、至少87％、至少88％、至少89％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、至少99％或100％，持续长达约1个月、长达约2个月、长达约2.5个月、长达约3个月、长达约3.5个月、长达约4个月、长达约4.5个月、长达约5个月、长达约5.5个月、长达约6个月、长达约6.5个月、长达约7个月、长达约7.5个月、长达约8个月、长达约8.5个月、长达约9个月、长达约9.5个月、长达约10个月、长达约10.5个月、长达约11个月、长达约11.5个月、长达约12个月、长达约12.5个月、长达约13个月、长达约13.5个月、长达约14个月、长达约14.5个月、长达约15个月、长达约15.5个月、长达约16个月、长达约16.5个月、长达约17个月、长达约17.5个月、长达约18个月、长达约18.5个月、长达约19个月、长达约19.5个月、长达约20个月、长达约20.5个月、长达约21个月、长达约21.5个月、长达约22个月、长达约22.5个月、长达约23个月、长达约23.5个月或长达约24个月或更长时间。

本申请所公开的组合物可以经由全身性途径或粘膜途径或透皮途径施用或直接施用到特定组织中。如本申请所用，术语“全身性施用”包括肠胃外施用途径。特别地，肠胃外施用包括皮下、腹膜内、静脉内、动脉内、肌内或胸骨内注射、静脉内或肾脏透析输注技术。通常，全身性肠胃外施用是肌内注射。如本申请所用，术语“粘膜施用”包括口服、鼻内、阴道内、直肠内、气管内、肠和眼部施用。优选地，施用是肌内的。

可以以单剂量方案或多剂量方案施用组合物。可以在初次免疫方案或在加强免疫方案中使用多剂量。在实施方案中，施用约1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29或30个剂量。在多剂量时间表中，可以通过相同或不同的途径施用各个剂量，例如肠胃外初次免疫和粘膜加强免疫、粘膜初次免疫和肠胃外加强免疫等。在各个方面，在第一个剂量后约2周、约3周、约4周、约5周、约6周、约2个月、约3个月、约4个月、约5个月、约6个月、约7个月、约8个月、约9个月、约10个月、约11个月、约12个月(1年)、约2年、约3年、约4年、约5年、约6年、约7年、约8年、约9年或约10年后施用加强剂量。在实施方案中，在施用初始剂量后每年施用加强剂量。在实施方案中，在施用先前剂量后3周或4周施用后续加强剂量。在实施方案中，在第0天施用第一个剂量，并且在第21天施用加强剂量。在实施方案中，在第0天施用第一个剂量，并且在第28天施用加强剂量。在实施方案中，在第0天施用第一个剂量，在第21天施用加强剂量，并且在施用第一个剂量后约六个月施用第二个加强剂量。在实施方案中，在第0天施用第一个剂量，并且在第28天施用加强剂量，并且在施用第一个剂量后约六个月施用第二个加强剂量。在实施方案中，在第0天施用第一个剂量，在第21天施用加强剂量，并且在施用第二个剂量后约六个月施用第二个加强剂量。在实施方案中，在第0天施用第一个剂量，并且在第28天施用加强剂量，并且在施用第二个剂量后约六个月施用第二个加强剂量。

在实施方案中，加强剂量包含与初始剂量相同的免疫组合物。在实施方案中，加强剂量包含与初始剂量不同的免疫组合物。在实施方案中，所述不同的免疫组合物是SARS-CoV-2刺突糖蛋白、编码SARS-CoV-2刺突糖蛋白的mRNA、编码SARS-CoV-2刺突糖蛋白的质粒DNA、编码SARS-CoV-2刺突糖蛋白的病毒载体或灭活的SARS-CoV-2病毒。在实施方案中，加强剂量包含初始组合物。在实施方案中，初始剂量包含SARS-CoV-2S糖蛋白(例如，具有SEQID NO:87的氨基酸序列的SARS-CoV-2S糖蛋白)，并且加强剂量包含相同的SARS-CoV-2S糖蛋白(例如，具有SEQ ID NO:87的氨基酸序列的SARS CoV-2S糖蛋白)。在实施方案中，初始剂量包含SARS-CoV-2S糖蛋白(例如，具有SEQ ID NO:87的氨基酸序列的SARS CoV-2S糖蛋白)，并且加强剂量包含不同的SARS-CoV-2S糖蛋白(例如，具有SEQ ID NO:132的氨基酸序列的SARS-CoV-2S糖蛋白)。在实施方案中，初始剂量包含SARS-CoV-2S糖蛋白(例如，具有SEQ ID NO:87的氨基酸序列的SARS CoV-2S糖蛋白和具有SEQ ID NO:132的氨基酸序列的SARS-CoV-2S糖蛋白)的组合。在实施方案中，加强剂量包含SARS-CoV-2S糖蛋白(例如，具有SEQ ID NO:87的氨基酸序列的SARS CoV-2S糖蛋白和具有SEQ ID NO:132的氨基酸序列的SARS CoV-2S糖蛋白)的组合。在实施方案中，初始剂量包含SARS-CoV-2S糖蛋白、编码SARS-CoV-2S糖蛋白的质粒DNA、编码SARS-CoV-2刺突糖蛋白的病毒载体或灭活的SARS-CoV-2病毒。在实施方案中，初始剂量包含SARS-CoV-2刺突糖蛋白、编码SARS-CoV-2刺突糖蛋白的质粒DNA、编码SARS-CoV-2刺突糖蛋白的病毒载体或灭活的SARS-CoV-2病毒，并且加强剂量包含一种或多种SARS-CoV-2S糖蛋白。

在实施方案中，如以μg测量的剂量可以是包括溶质的剂量的总重量、或CoV S多肽纳米颗粒的重量、或CoV S多肽的重量。使用蛋白质浓度测定(A280或ELISA)来测量剂量。

包括用于小儿施用的抗原剂量可以在约5μg至约25μg、约1μg至约300μg、约90μg至约270μg、约100μg至约160μg、约110μg至约150μg、约120μg至约140μg或约140μg至约160μg的范围内。在实施方案中，所述剂量为约120μg，与明矾一起施用。在各个方面，小儿剂量可以在约1μg至约90μg的范围内。在实施方案中，CoV刺突(S)多肽的剂量为约1μg、约2μg、约3μg、约4μg、约5μg、约6μg、约7μg、约8μg、约9μg、约10μg、约11μg、约12μg、约13μg、约14μg、约15μg、约16μg、约17μg、约18μg、约19μg、约20μg、约21、约22、约23、约24、约25μg、约26μg、约27μg、约28μg、约29μg、约30μg、约40μg、约50、约60、约70、约80、约90约100μg、约110μg、约120μg、约130μg、约140μg、约150μg、约160μg、约170μg、约180μg、约190μg、约200μg、约210μg、约220μg、约230μg、约240μg、约250μg、约260μg、约270μg、约280μg、约290μg或约300μg，包括其间的所有值和范围。在实施方案中，CoV S多肽的剂量为5μg。在实施方案中，CoV S多肽的剂量为25μg。在实施方案中，CoV S多肽的剂量对于初始剂量和加强剂量是相同的。在实施方案中，CoV S多肽的剂量对于初始剂量和加强剂量是不同的。

可以在有或没有佐剂的情况下向某些人群施用。在某些方面，组合物可以不含添加的佐剂。在此类情况下，可以将剂量增加约10％。

在实施方案中，与非天然存在的CoV S多肽一起施用的佐剂的剂量为约1μg至约100μg，例如约1μg、约2μg、约3μg、约4μg、约5μg、约6μg、约7μg、约8μg、约9μg、约10μg、约11μg、约12μg、约13μg、约14μg、约15μg、约16μg、约17μg、约18μg、约19μg、约20μg、约21μg、约22μg、约23μg、约24μg、约25μg、约26μg、约27μg、约28μg、约29μg、约30μg、约31μg、约32μg、约33μg、约34μg、约35μg、约36μg、约37μg、约38μg、约39μg、约40μg、约41μg、约42μg、约43μg、约44μg、约45μg、约46μg、约47μg、约48μg、约49μg、约50μg、约51μg、约52μg、约53μg、约54μg、约55μg、约56μg、约57μg、约58μg、约59μg、约60μg、约61μg、约62μg、约63μg、约64μg、约65μg、约66μg、约67μg、约68μg、约69μg、约70μg、约71μg、约72μg、约73μg、约74μg、约75μg、约76μg、约77μg、约78μg、约79μg、约80μg、约81μg、约82μg、约83μg、约84μg、约85μg、约86μg、约87μg、约88μg、约89μg、约90μg、约91μg、约92μg、约93μg、约94μg、约95μg、约96μg、约97μg、约98μg、约99μg或约100μg的佐剂。在实施方案中，佐剂的剂量为约50μg。在实施方案中，所述佐剂是皂苷佐剂，例如MATRIX-M^TM。

在实施方案中，施用的剂量的体积为约0.1mL至约1.5mL，例如约0.1mL、约0.2mL、约0.25mL、约0.3mL、约0.4mL、约0.5mL、约0.6mL、约0.7mL、约0.8mL、约0.9mL、约1.0mL、约1.1mL、约1.2mL、约1.3mL、约1.4mL或约1.5mL。在实施方案中，施用的剂量的体积为0.25mL。在实施方案中，施用的剂量的体积为0.5mL。在实施方案中，施用的剂量的体积为0.6mL。

在针对MERS、SARS或SARS-CoV-2冠状病毒的疫苗的特定实施方案中，所述剂量可以包含以下浓度的CoV S多肽：约1μg/mL至约50μg/mL、10μg/mL至约100μg/mL、约10μg/mL至约50μg/mL、约175μg/mL至约325μg/mL、约200μg/mL至约300μg/mL、约220μg/mL至约280μg/mL或约240μg/mL至约260μg/mL。

在另一个实施方案中，本申请公开文本提供了配制疫苗组合物的方法，所述疫苗组合物诱导针对哺乳动物的感染或其至少一种疾病症状的免疫，所述方法包括向所述组合物中添加有效剂量的纳米颗粒或CoV S多肽。所公开的CoV S多肽和纳米颗粒可用于制备刺激免疫反应的组合物，所述免疫反应赋予针对感染原的免疫或实质性免疫。因此，在一个实施方案中，本申请公开文本提供了在受试者中诱导针对感染或其至少一种疾病症状的免疫的方法，其方法包括施用至少一个有效剂量的纳米颗粒和/或CoV S多肽。

在实施方案中，将所述CoV S多肽或包含所述多肽的纳米颗粒与另外的免疫原性组合物组合施用。在实施方案中，所述另外的免疫原性组合物诱导针对SARS-CoV-2的免疫反应。在实施方案中，将所述另外的免疫原性组合物在所公开的CoV S多肽或包含所述多肽的纳米颗粒的约1分钟、约5分钟、约10分钟、约20分钟、约30分钟、约40分钟、约50分钟、约1小时、约2小时、约3小时、约4小时、约5小时、约6小时、约7小时、约8小时、约9小时、约10小时、约11小时、约12小时、约13小时、约14小时、约15小时、约16小时、约17小时、约18小时、约19小时、约20小时、约21小时、约22小时、约23小时、约1天、约2天、约3天、约4天、约5天、约6天、约7天、约8天、约9天、约10天、约11天、约12天、约13天、约14天、约15天、约16天、约17天、约18天、约19天、约20天、约21天、约22天、约23天、约24天、约25天、约26天、约27天、约28天、约29天、约30天或约31天内施用。在实施方案中，将所述另外的组合物与第一个剂量的包含CoV S多肽或含有所述多肽的纳米颗粒的组合物一起施用。在实施方案中，将所述另外的组合物与加强剂量的包含CoV S多肽或含有所述多肽的纳米颗粒的组合物一起施用。

在实施方案中，所述另外的免疫原性组合物包含编码SARS-CoV-2刺突糖蛋白的mRNA、编码SARS-CoV-2刺突糖蛋白的质粒DNA、编码SARS-CoV-2刺突糖蛋白的病毒载体或灭活的SARS-CoV-2病毒。

在实施方案中，所述另外的免疫原性组合物包含编码CoV S多肽的mRNA。在实施方案中，所述mRNA编码在SEQ ID NO:1的位置986和987处包含脯氨酸取代的CoV S多肽。在实施方案中，所述mRNA编码包含完整弗林蛋白酶切割位点的CoV S多肽。在实施方案中，所述mRNA编码包含在SEQ ID NO:1的位置986和987处的脯氨酸取代和完整弗林蛋白酶切割位点的CoV S多肽。在实施方案中，所述mRNA编码包含在SEQ ID NO:1的位置986和987处的脯氨酸取代和无活性的弗林蛋白酶切割位点的CoV S多肽。在实施方案中，所述mRNA编码具有SEQ ID NO:87的氨基酸序列的CoV S多肽。在实施方案中，编码CoV S多肽的mRNA被封装在脂质纳米颗粒中。包含编码CoV S多肽的mRNA的示例性免疫原性组合物描述于Jackson等人N.Eng.J.Med.2020(针对SARS-CoV-2的mRNA疫苗初步报告)中，将其通过引用以其整体并入本申请。在实施方案中，将包含编码CoV S多肽的mRNA的组合物以25μg、100μg或250μg的剂量施用。

在实施方案中，所述另外的免疫原性组合物包含编码CoV S多肽的腺病毒载体。在实施方案中，所述AAV载体编码野生型CoV S多肽。在实施方案中，所述AAV载体编码包含在SEQ ID NO:1的位置986和987处的脯氨酸取代和完整弗林蛋白酶切割位点的CoV S多肽。在实施方案中，所述AAV载体编码包含在SEQ ID NO:1的位置986和987处的脯氨酸取代和无活性的弗林蛋白酶切割位点的CoV S多肽。在实施方案中，所述AAV载体编码具有SEQ ID NO:87的氨基酸序列的CoV S多肽。以下出版物描述了包含编码CoV S多肽的腺病毒载体的免疫原性组合物，将其每一个通过引用以其整体并入本申请：van Doremalen N.等人Asingledose of ChAdOx1 MERS provides protective immunity in rhesusmacaques.Science Advances,2020；van Doremalen N.等人ChAdOx1 nCoV-19vaccinationprevents SARS-CoV-2pneumonia in rhesus macaques.bioRxiv,(2020)。

在实施方案中，所述另外的免疫原性组合物包含脱氧核糖核酸(DNA)。在实施方案中，所述另外的免疫原性组合物包含质粒DNA。在实施方案中，所述质粒DNA编码CoV S多肽。在实施方案中，所述DNA编码包含在SEQ ID NO:1的位置986和987处的脯氨酸取代和完整弗林蛋白酶切割位点的CoV S多肽。在实施方案中，所述DNA编码包含在SEQ ID NO:1的位置986和987处的脯氨酸取代和无活性的弗林蛋白酶切割位点的CoV S多肽。在实施方案中，所述DNA编码具有SEQ ID NO:87的氨基酸序列的CoV S多肽。

在实施方案中，所述另外的免疫原性组合物包含灭活的病毒疫苗。

在实施方案中，将CoV S多肽或包含CoV S多肽的纳米颗粒施用至已经或先前已经具有证实的由SARS-CoV-2或异源SARS-CoV-2毒株引起的感染的患者。感染SARS-CoV-2或异源SARS-CoV-2毒株可以通过核酸扩增测试(例如，聚合酶链式反应)或血清学测试(例如，针对抗SARS-CoV-2病毒抗原的抗体的测试)来证实。在实施方案中，在患者被诊断为患有COVID-19后至少约3天、至少约1周、至少约2周、至少约3周、至少约4周，向患者施用CoV S多肽或包含CoV S多肽的纳米颗粒。在实施方案中，在患者被诊断为患有COVID-19后1周与1年之间，例如约1周、约2周、约3周、约4周、约5周、约6周、约1个月、约2个月、约3个月、约4个月、约5个月、约6个月、约7个月、约8个月、约9个月、约10个月、约11个月或约1年，向患者施用CoV S多肽或包含CoV S多肽的纳米颗粒。在实施方案中，在患者被诊断为患有COVID-19后1周与20年之间，例如约1周、约2周、约3周、约4周、约5周、约6周、约1个月、约2个月、约3个月、约4个月、约5个月、约6个月、约7个月、约8个月、约9个月、约10个月、约11个月、约1年、约2年、约3年、约4年、约5年、约6年、约7年、约8年、约9年、约10年、约11年、约12年、约13年、约14年、约15年、约16年、约17年、约18年、约19年或约20年，向患者施用CoV S多肽或包含CoV S多肽的纳米颗粒。

在实施方案中，在患者已被施用第一免疫原性组合物之后施用CoV S多肽或包含其的纳米颗粒。第一免疫原性组合物的非限制性例子包括SARS-CoV-2刺突糖蛋白、编码SARS-CoV-2刺突糖蛋白的mRNA、编码SARS-CoV-2刺突糖蛋白的质粒DNA、编码SARS-CoV-2刺突糖蛋白的病毒载体或灭活的SARS-CoV-2病毒。在实施方案中，在施用第一免疫原性组合物后约1周与约1年之间、约1周与1个月之间、约3周与4周之间、约1周与5年之间、约1年与约5年之间、约1年与约3年之间、约3年与约5年之间、约5年与约10年之间、约1年与约10年之间或约1年与约2年之间，施用CoV S多肽或包含其的纳米颗粒。在实施方案中，在施用第一免疫原性组合物后约1周与约1年之间，例如在施用第一免疫原性组合物后约1周、约2周、约3周、约4周、约5周、约6周、约7周、约8周、约9周、约10周、约1个月、约2个月、约3个月、约4个月、约5个月、约6个月、约7个月、约8个月、约9个月、约10个月、约11个月或约1年，施用CoV S多肽或包含其的纳米颗粒。

在实施方案中，所述CoV S蛋白或包含CoV S蛋白的纳米颗粒可用于制备免疫原性组合物以刺激赋予针对MERS、SARS、SARS-CoV-2和异源SARS-CoV-2毒株中的一种或多种的免疫力或实质性免疫的免疫反应。粘膜免疫力和细胞免疫力都可以有助于针对感染和疾病的免疫。上呼吸道局部分泌的抗体是抵抗天然感染的主要因子。分泌型免疫球蛋白A(sIgA)参与保护上呼吸道，并且血清IgG参与保护下呼吸道。感染诱导的免疫反应预防相同病毒或抗原性相似的病毒株再感染。用本申请所公开的纳米颗粒免疫后在宿主中产生的抗体也可以施用于其他宿主，从而在受试者中提供被动施用。

在实施方案中，所述CoV S蛋白或包含CoV S蛋白的纳米颗粒诱导针对含有具有选自以下的一个或多个修饰的S蛋白的SARS-CoV-2病毒的交叉中和抗体：

(a)NTD的一个或多个氨基酸的缺失，其中所述一个或多个氨基酸选自氨基酸56、57、131、132、229、230、231或其组合；以及

(b)NTD的一个或多个氨基酸的突变，其中所述一个或多个突变选自氨基酸67、82、133、229、202、209、240、139、5、233、7、13、125、177或其组合；

(c)RBD的一个或多个氨基酸的突变，其中所述一个或多个突变选自氨基酸488、404、471、464、439、481、426、440及其组合；

(d)SD1/2的一个或多个氨基酸的突变，其中所述一个或多个氨基酸选自601、557、668、642及其组合；

(e)无活性的弗林蛋白酶切割位点(对应于氨基酸669-672中的一个或多个突变)；

(f)S2亚基的一个或多个氨基酸的缺失，其中所述氨基酸选自676-702、702-711、775-793、806-815及其组合；

(g)S2亚基的一个或多个氨基酸的突变，其中所述氨基酸选自973、974、703、1105、688、969、1014和1163及其组合；

(h)从TMCT(氨基酸1201-1260)中的一个或多个氨基酸的缺失，其中所述CoV S糖蛋白的氨基酸关于SEQ ID NO:2进行编号。

在实施方案中，所述CoV S蛋白或包含CoV S蛋白的纳米颗粒诱导针对含有具有选自以下的一个或多个修饰的S蛋白的SARS-CoV-2病毒的交叉中和抗体：氨基酸56的缺失、氨基酸57的缺失、氨基酸131的缺失、N488Y、A557D、D601G、P668H、T703I、S969A、D1105H、N426K和Y440F，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。

在实施方案中，所述CoV S蛋白或包含CoV S蛋白的纳米颗粒诱导针对含有具有选自以下的一个或多个修饰的S蛋白的SARS-CoV-2病毒的交叉中和抗体：氨基酸56的缺失、氨基酸57的缺失、氨基酸131的缺失、N488Y、A557D、D601G、P668H、T703I、S969A和D1105H，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。

在实施方案中，所述CoV S蛋白或包含CoV S蛋白的纳米颗粒诱导针对含有具有选自以下的一个或多个修饰的S蛋白的SARS-CoV-2病毒的交叉中和抗体：D67A、D202G、L229H、K404N、E471K、N488Y、D601G和A688V，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。

在实施方案中，所述CoV S蛋白或包含CoV S蛋白的纳米颗粒诱导针对含有具有选自以下的一个或多个修饰的S蛋白的SARS-CoV-2病毒的交叉中和抗体：氨基酸229-231的缺失、D67A、D202G、K404N、E471K、N488Y、D601G和A688V，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。

在实施方案中，所述CoV S蛋白或包含CoV S蛋白的纳米颗粒诱导针对含有具有选自以下的一个或多个修饰的S蛋白的SARS-CoV-2病毒的交叉中和抗体：氨基酸229-231的缺失、L5F、D67A、D202G、K404N、E471K、N488Y、D601G和A688V，其中所述氨基酸关于具有SEQ IDNO:2的氨基酸序列的CoV S多肽进行编号。

在实施方案中，所述CoV S蛋白或包含CoV S蛋白的纳米颗粒诱导针对含有具有选自以下的一个或多个修饰的S蛋白的SARS-CoV-2病毒的交叉中和抗体：L5F、T7N、P13S、D125Y、R177S、K404T、E471K、N488Y、D601G、H642Y、T1014I和V1163F，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。

在实施方案中，所述CoV S蛋白或包含CoV S蛋白的纳米颗粒诱导针对具有包含选自以下的一个或多个修饰的S蛋白的SARS-CoV-2病毒的交叉中和抗体：W139C和L439R，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，包含W139C和L439R修饰的CoV S蛋白与具有SEQ ID NO:117或SEQ ID NO:5的氨基酸序列的信号肽一起表达。在实施方案中，所述CoV S蛋白或包含CoV S蛋白的纳米颗粒诱导针对具有选自以下的一个或多个修饰的SARS-CoV-2病毒的交叉中和抗体：D601G、W139C和L439R，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，包含D601G、W139C和L439R修饰的CoV S蛋白或纳米颗粒与具有SEQ ID NO:117或SEQ IDNO:5的氨基酸序列的信号肽一起表达。

在实施方案中，所述CoV S蛋白或包含CoV S蛋白的纳米颗粒诱导针对具有选自以下的一个或多个修饰的SARS-CoV-2病毒的交叉中和抗体：D601G、L5F、D67A、D202G、氨基酸229-231的缺失、R233I、K404N、E471K、N488Y和A688V，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。在实施方案中，所述CoV S蛋白或包含CoV S蛋白的纳米颗粒诱导针对具有选自以下的一个或多个修饰的SARS-CoV-2病毒的交叉中和抗体：L5F、D67A、D202G、氨基酸229-231的缺失、R233I、K404N、E471K、N488Y和A688V，其中所述氨基酸关于具有SEQ ID NO:2的氨基酸序列的CoV S多肽进行编号。

在实施方案中，本申请公开文本提供了产生高亲和力抗MERS-CoV、抗SARS-CoV和抗SARS-CoV-2病毒抗体中的一种或多种的方法。通过用本申请所公开的纳米颗粒免疫而产生的高亲和力抗体通过以下方式来产生：向动物施用包含S CoV多肽或含有S CoV多肽的纳米颗粒的免疫原性组合物，从所述动物中收集血清和/或血浆，并且从血清/和或血浆中纯化抗体。在一个实施方案中，所述动物是人。在实施方案中，所述动物是鸡、小鼠、豚鼠、大鼠、兔、山羊、人、马、羊或牛。在一个实施方案中，所述动物是牛或马。在另一个实施方案中，所述牛或马动物是转基因的。在又另一个实施方案中，所述转基因牛或马动物产生人抗体。在实施方案中，所述动物产生单克隆抗体。在实施方案中，所述动物产生多克隆抗体。在一个实施方案中，所述方法还包括施用佐剂或免疫刺激化合物。在另一个实施方案中，向人类受试者施用经纯化的高亲和力抗体。在一个实施方案中，所述人类受试者有感染MERS、SARS和SARS-CoV-2中的一种或多种的风险。

在实施方案中，将所述CoV S蛋白或纳米颗粒与流感糖蛋白或包含流感糖蛋白的纳米颗粒共同施用。合适的糖蛋白和纳米颗粒描述于美国公开号2018/0133308和美国公开号2019/0314487中，将其中的每个公开案通过引用以其整体并入本申请。在实施方案中，将所述CoV S蛋白或纳米颗粒与以下各项共同施用：(a)洗涤剂-核心纳米颗粒，其中所述洗涤剂-核心纳米颗粒包含来自B型流感毒株的重组流感血凝素(HA)糖蛋白；和(b)血凝素皂苷基质纳米颗粒(HaSMaN)，其中所述HaSMaN包含来自A型流感毒株的重组流感HA糖蛋白和ISCOM基质佐剂。在实施方案中，将所述CoV S蛋白或纳米颗粒与包含非离子型洗涤剂核心和流感HA糖蛋白的纳米颗粒共同施用，其中所述流感HA糖蛋白含有从非离子型洗涤剂核心向外突出的头部区域和与非离子型洗涤剂核心缔合的跨膜结构域，其中所述流感HA糖蛋白是HA0糖蛋白，其中所述流感HA糖蛋白的氨基酸序列与天然流感HA蛋白的氨基酸序列具有100％同一性。在实施方案中，将所述流感糖蛋白或纳米颗粒与所述CoV S蛋白或纳米颗粒共同配制。

将本申请公开文本中引用的所有专利、专利申请、参考文献和期刊文章出于所有目的通过引用以其整体明确地并入本申请。

实例

实例1

冠状病毒刺突(S)多肽纳米颗粒的表达和纯化

已在杆状病毒表达系统中表达了天然冠状病毒刺突(S)多肽(SEQ ID NO:1和SEQID NO:2)和CoV刺突多肽(其具有对应于SEQ ID NO:3、4、38、41、44、48、51、54、58、61、63、65、67、73、75、78、79、82、83、85、87、106、108、89、112-115、132、133、114、138、141、144、147、151、153、156、158、164-168的氨基酸序列)，并且挑选并确认了表达冠状病毒刺突(S)多肽的重组噬斑。在每种情况下，信号肽是SEQ ID NO:5。图4和图9显示了CoV刺突多肽BV2364、BV2365、BV2366、BV2367、BV2368、BV2369、BV2373、BV2374和BV2375的成功纯化。表2显示了上述CoV刺突多肽的序列特征。

表2：选定的CoV刺突多肽

野生型BV2361蛋白(SEQ ID NO:2)与人血管紧张素转化酶2前体(hACE2)结合。进行了生物膜层干涉测量术和ELISA，以评估CoV S多肽的结合。

生物膜层干涉测量术(BLI)

使用Octet QK384系统(Pall FortéBio，加利福尼亚州弗里蒙特)进行了BLI实验。将His标记的人ACE2(2μg mL-1)固定在带镍的Ni-NTA生物传感器尖端上。在基线后，将含SARS-CoV-2S蛋白的样品连续稀释2倍，并且允许其缔合600秒，然后再使其解离900秒。使用Octet软件HT 101:1全局曲线拟合分析数据。

CoV S多肽BV2361、BV2365、BV2369、BV2365、BV2373、BV2374保留与hACE2结合的能力(图5、图11A-图11C)。解离动力学表明，在不存在流体相S蛋白的情况下，如通过观察900秒的最小解离或无解离所证实的，S蛋白保持紧密结合(图11A-图11C)。

此外，结合是特异性的。野生型CoV S蛋白BV2361以及CoV S多肽BV2365和BV2373不结合MERS-CoV受体二肽基肽酶IV(DPP4)。此外，MERS S蛋白不与人血管紧张素转化酶2前体(hACE2)结合(图6和图11D-图11F)。

ELISA

通过ELISA确认了CoV S多肽对hACE2的特异性。将九十六孔板在4℃下用100μLSARS-CoV-2刺突蛋白(2μg/mL)包被过夜。用含有0.05％Tween(PBS-T)缓冲液的磷酸盐缓冲盐水洗涤板，并将其用TBS Startblock封闭缓冲液(ThermoFisher,Scientific)封闭。将His标记的hACE2和hDPP4受体连续稀释3倍(5-0.0001μg mL-1)，并且在室温下将其添加到包被的孔中持续2小时。用PBS-T洗涤板。添加最适稀释的辣根过氧化物酶(HRP)缀合的抗组氨酸，并且通过添加3,3’,5,5’-四甲基联苯胺过氧化物酶底物(TMB，T0440-IL，Sigma，美国密苏里州圣路易斯)来显色。用SpectraMax Plus板读取器(Molecular Devices，美国加利福尼亚州桑尼维尔)在450nm的OD下读取板，并且用SoftMax软件分析数据。使用GraphPadPrism 7.05软件通过4参数拟合来计算EC50值。

ELISA结果表明，野生型CoV S多肽(BV2361)、BV2365和BV2373蛋白特异性地结合hACE2，但未能结合由MERS-CoV使用的hDPP-4受体(IC₅₀>5000ng mL-1)。野生型CoV S多肽和BV2365以相似的亲和力结合至hACE2(IC₅₀＝36-38ng/mL)，而BV2373在低2倍的浓度下达到50％的hACE2结合的饱和(IC₅₀＝18ng/mL)(图7、图11D-图11F)。

蛋白质和纳米颗粒产生

通过感染Sf9昆虫细胞来扩增重组病毒。在约3MOI(感染复数＝病毒ffu或pfu/细胞)下用杆状病毒感染昆虫细胞培养物。感染后48-72小时收获培养物和上清液。将大约30mL的粗细胞收获物通过以大约800x g离心15分钟进行澄清。如下所述，将所得的含有冠状病毒刺突(S)蛋白的粗细胞收获物纯化为纳米颗粒。

为了产生纳米颗粒，将非离子型表面活性剂壬基酚乙氧基化物NP-9用于膜蛋白提取方案中。经由通过阴离子交换层析、小扁豆凝集素亲和/HIC和阳离子交换层析进一步纯化粗提取物。通过洗涤剂处理使洗涤过的细胞裂解，然后对其进行低pH处理，这导致BV和Sf9宿主细胞DNA和蛋白质沉淀。使经中和的低pH处理裂解液澄清，并且在进行第二次低pH处理之前在阴离子交换和亲和层析上进一步纯化。

使用亲和层析来去除Sf9/BV蛋白、DNA和NP-9，并且浓缩冠状病毒刺突(S)蛋白。简言之，小扁豆凝集素是一种含有钙和锰的金属蛋白，其可逆地结合含有葡萄糖或甘露糖的多糖和糖基化蛋白。将含冠状病毒刺突(S)蛋白的阴离子交换流过级分加载到小扁豆凝集素亲和层析树脂(Capto小扁豆凝集素，GE Healthcare)上。糖基化冠状病毒刺突(S)蛋白选择性地结合至树脂，而非糖基化蛋白和DNA在柱流过物中被去除。通过含有高盐和低摩尔浓度的甲基α-D-吡喃甘露糖苷(MMP)的缓冲液去除弱结合的糖蛋白。

还使用柱洗涤物来将NP-9洗涤剂用表面活性剂聚山梨醇酯80(PS80)进行洗涤剂交换。用高浓度的MMP将冠状病毒刺突(S)多肽以纳米颗粒结构从小扁豆凝集素柱中洗脱。在洗脱后，将冠状病毒刺突(S)蛋白三聚体组装成由包含在洗涤剂核心中的冠状病毒刺突(S)蛋白三聚体和PS80构成的纳米颗粒。

实例2

冠状病毒刺突(S)多肽纳米颗粒疫苗在小鼠中的免疫原性

使用雌性BALB/c小鼠(7-9周龄；Harlan Laboratories Inc.，马里兰州弗雷德里克)评价了如实例1所述的包含SEQ ID NO:87的CoV S多肽(也称为“BV2373”)的冠状病毒刺突(S)蛋白组合物在鼠类模型中的免疫原性和毒性。在存在和不存在皂苷佐剂(例如，MATRIX-M^TM)的情况下评价了组合物。含有MATRIX-M^TM的组合物含有5μg的MATRIX-M^TM。将含有不同剂量(包括0.01μg、0.1μg、1μg和10μg)的冠状病毒刺突(S)多肽的疫苗作为单剂量(也称为单次初免剂量)(研究第14天)或作为间隔14天(研究第0天和第14天)的两个剂量(也称为初次免疫/加强免疫方案)肌内施用。安慰剂组用作非免疫对照。在研究第-1天、第13天、第21天和第28天收集血清进行分析。在一次(单剂量)或两次(两个剂量)免疫后42天，用SARS-CoV-2鼻内攻击接种疫苗的动物和对照动物。

疫苗免疫原性

用0.1-10μg BV2373和MATRIX-M^TM的单次初免剂量免疫的动物具有升高的抗S IgG滴度，其在单次免疫后21-28天检测到(图13B)。用10μg剂量的BV2373和MATRIX-M^TM免疫的小鼠产生了阻断hACE2受体与CoV S蛋白结合的抗体和在单次初免剂量后21-28天检测到的病毒中和抗体(图14和图15)。用初次免疫/加强免疫方案(两个剂量)免疫的动物具有显著升高的抗S IgG滴度，其在所有剂量水平下在加强免疫后7-16天均检测到(图13A)。用BV2373(1μg和10μg)和MATRIX-M^TM免疫的动物在免疫后具有相似的高抗S IgG滴度(分别地，GMT＝139,000和84,000)。与在没有佐剂的情况下用10μg BV2373免疫的小鼠相比，用BV2373(0.1μg、1μg或10μg)和MATRIX-M^TM免疫的小鼠具有显著(p≤0.05和p≤0.0001)更高的抗S IgG滴度(图13A)。这些结果表明了由MATRIX-M^TM佐剂提供10倍至100倍剂量节省的可能性。此外，用两个剂量的BV2373和MATRIX-M^TM免疫产生了高滴度抗体，其在所有剂量水平下均阻断hACE2受体与S蛋白结合(IC50＝218-1642)并中和SARS-CoV-2对Vero E6细胞的致细胞病变效应(CPE)(CPE的100％阻断＝7680-20,000)(图14和图15)。

SARS CoV-2攻击

为了评价保护性免疫的诱导，用SARS-CoV-2攻击了免疫的小鼠。由于小鼠不支持野生型SARS-CoV-2病毒的复制，因此在初始疫苗接种后第52天，用表达hACE2的腺病毒(Ad/hACE2)鼻内感染小鼠以允许其复制。向小鼠鼻内接种1.5x 10⁵pfu的SARS-CoV-2，在各鼻孔之间分50μL。在感染当天称量受攻击的小鼠，并且在感染后每天称重持续7天。在感染后4天和7天，处死来自疫苗接种组和对照组各自的5只小鼠，并且收获肺并准备进行肺组织学检查。

通过噬斑测定对病毒滴度进行了定量。简言之，使用1.0mm玻璃珠(SigmaAldrich)和Beadruptor(Omini International Inc.)在PBS中使所收获的肺均质化。将匀浆添加到Vero E6近汇合培养物中，并且使用6点稀释曲线通过计数噬斑形成单位(pfu)来测定SARS-CoV-2病毒滴度。

在感染后4天，经安慰剂治疗的小鼠具有10⁴SARS-CoV-2pfu/肺，而在没有MATRIX-M^TM的情况下用BV2363免疫的小鼠具有10³pfu/肺(图16)。仅BV2373加MATRIX-M^TM初次免疫的小鼠展现出病毒滴度的剂量依赖性降低，其中在感染后第4天，10μg BV2373剂量的接受者没有可检测的病毒。与接种安慰剂的小鼠相比，接受1μg、0.1μg和0.01μg BV2373剂量的小鼠均显示出滴度显著降低。在初次免疫/加强免疫组中，用10μg、1μg和0.1μg剂量免疫的小鼠具有几乎不可检测的肺病毒载量，而0.01μg组相对于安慰剂动物显示降低1个对数的降低。

体重减轻与病毒载量的发现结果平行。与未接种疫苗的安慰剂动物相比，接受单剂量的BV2373(0.1μg、1μg和10μg)和MATRIX-M^TM的动物显示出显著预防体重减轻(图17A)。在所有剂量水平下，接受初免和加强剂量加佐剂的小鼠也均展现出显著预防体重减轻(图17B-图17C)。评价了佐剂的存在对预防体重减轻的作用。相对于安慰剂，接受初次免疫/加强免疫(两个剂量)+佐剂的小鼠显著被保护免于体重减轻，而在没有佐剂的情况下免疫的组则没有(图17C)。这些结果表明，BV2373赋予预防SARS-CoV-2，并且与较低的血清学反应相关的低剂量疫苗不会加剧体重减轻或展现出疾病加重。

在感染后第4天和第7天评价了肺组织病理学(图18A和图18B)。在感染后第4天，经安慰剂免疫的小鼠显示出在大气道中的上皮细胞剥蚀以及由混合炎性细胞群包围的肺泡隔增厚。在整个肺中观察到外周小动脉成套有炎性细胞，其主要由嗜中性粒细胞和巨噬细胞组成。到感染后第7天，经安慰剂治疗的小鼠显示细支气管周炎症和外周小动脉成套增加。肺泡隔增厚在整个肺泡隔中仍伴有弥漫性间质性炎症增加(图18B)。

经BV2373免疫的小鼠在感染后第4天和第7天都显示出肺部病理以剂量依赖性方式显著降低。与安慰剂小鼠相比，仅初次免疫组在10μg和1μg剂量下显示炎症减少，其中支气管和小动脉周围炎症减少。在较低剂量的仅初次免疫组中，肺部炎症类似于安慰剂组，与体重减轻和肺部病毒滴度相关。对于所有测试剂量，初次免疫/加强免疫免疫组都显示肺部炎症显著降低，这再次与肺部病毒滴度和体重减轻数据相关。在第4天和第7天，基本上保留大支气管和小支气管中的上皮细胞，其中细支气管脱落和病毒感染迹象最小。用10μg、1μg和0.1μg剂量免疫的动物的小动脉具有最少的炎症，其中在0.01μg剂量的情况下仅观察到适度的成套，类似于安慰剂。在接受较高剂量的动物中肺泡炎症减少，其中仅较低的0.01μg剂量与炎症相关(图18A-图18B)。这些数据证明，BV2373减少攻击后的肺部炎症，并且甚至引起最小或没有引起可检测的中和活性的BV2373的剂量和方案也与对病毒的炎性反应加剧不相关。此外，疫苗不会在受攻击的小鼠中引起疫苗相关的增强型呼吸系统疾病(VAERD)。

T细胞反应

评价了包含SEQ ID NO:87的CoV S多肽的疫苗组合物对T细胞反应的影响。以间隔21天的2个剂量用10μg BV2373加或不加5μg MATRIX-M^TM肌内免疫BALB/c小鼠(每组N＝6)。在第二次免疫后7天(研究第28天)收集脾脏。未接种疫苗的组(N＝3)用作对照。

通过ELISPOT^TM酶联免疫吸附测定和细胞内细胞因子染色(ICCS)测量来自第二次免疫后7天(研究第28天)收集的脾脏的抗原特异性T细胞反应。与单独的BV2373相比，在用BV2373和MATRIX-M^TM免疫的小鼠的脾脏中，离体刺激后IFN-γ分泌性细胞的数量增加了20倍(p＝0.002)，如通过ELISPOT^TM测定所测量的(图19)。为了分开检查CD4+和CD8+T细胞反应，与表面标记物染色组合进行了ICCS测定。所示出的数据在CD44hi CD62L-效应记忆T细胞群上进行门控。与在没有佐剂的情况下免疫的小鼠相比，在来自用BV2373免疫的小鼠的脾脏中IFN-γ+、TNF-α+和IL-2+细胞因子分泌性CD4+和CD8+T细胞的频率显著更高(p<0.0001)(图20A-图20C和图21A-图21C)。此外，与在不存在佐剂的情况下免疫的小鼠相比，在来自经BV2373/MATRIX-M^TM免疫的小鼠的脾脏中同时产生至少两种或三种细胞因子的多功能性CD4+和CD8+T细胞的频率也显著增加(p<0.0001)(图20D-图20E和图21D-图21E)。用BV2373/MATRIX-M^TM免疫在CD4+和CD8+T细胞群内都产生了更高比例的多功能表型(例如，分泌IFN-γ、TNF-α和IL-2中的多于一种的T细胞)。在记忆CD4+T细胞中检测到的多菜单型的比例高于在CD8+T细胞中的比例(图22)。

还分别通过ICCS和ELISPOT^TM测定了CD4+T细胞的2型细胞因子IL-4和IL-5分泌。与单独用BV2373免疫相比，用BV2373/MATRIX-M^TM免疫还增加了2型细胞因子IL-4和IL-5分泌(2倍)，但程度小于1型细胞因子产生的增强(例如IFN-γ增加20倍)(图23A-图23C)。这些结果表明，MATRIX-M^TM佐剂的施用使CD4+T细胞偏向产生Th1反应。

通过测量脾脏中CD4+滤泡辅助性T(TFH)细胞和生发中心(GC)B细胞的频率来评估免疫对生发中心形成的影响。MATRIX-M^TM施用显著增加了脾脏中TFH细胞(CD4+CXCR5+PD-1+)的频率(p＝0.01)以及GC B细胞(CD19+GL7+CD95+)的频率(p＝0.0002)(图24A-图24B和图25A-图25B)。

实例3

冠状病毒刺突(S)多肽纳米颗粒疫苗在橄榄狒狒中的免疫原性

评估了包含BV2373的疫苗组合物在狒狒中的免疫原性。通过肌内(IM)注射以间隔21天的两个剂量施用一系列剂量(1μg、5μg和25μg)的BV2373和50μg MATRIX-M^TM佐剂来免疫成年橄榄狒狒。为了评估MATRIX-M^TM在非人灵长类动物中的辅助活性，在没有MATRIX-M^TM的情况下用25μg的BV2373免疫另一组动物。在所有剂量水平下，在用BV2373/MATRIX-M^TM免疫的动物中，在单次初次免疫的21天内均检测到抗S蛋白IgG滴度(GMT＝1249-19,000)。在所有剂量水平下，加强免疫后1至2周(第28天和第35天)内，抗S蛋白IgG滴度均增加超过一个对数(GMT＝33,000-174,000)。(图26A)。

用BV2373(5μg或25μg)和MATRIX-M^TM单次免疫后，在动物中检测到低水平的hACE2受体阻断抗体(GMT＝22-37)。在用BV2373/MATRIX-M^TM免疫的所有组中，在加强免疫的一至二周内，受体阻断抗体滴度均显著增加(GMT＝150-600)(图26B)。在用BV2373/MATRIX-M^TM单次免疫后，在所有剂量组中，病毒中和抗体均升高(GMT＝190-446)。单独用25μg BV2373免疫的动物没有可检测的阻断S蛋白与hACE2结合的抗体(图26C)。加强免疫后一周，中和滴度增加6倍至8倍(GMT＝1160-3846)。第二次免疫后，中和滴度再增加25倍至38倍(GMT＝6400-17,000)(图26C)。抗S IgG水平与中和抗体滴度之间存在显著相关性(p<0.0001)(图27)。佐剂化疫苗在非人灵长类动物中的免疫原性与实例2的结果一致，并且进一步支持MATRIX-M^TM在促进中和抗体的产生和剂量节省中的作用。

在第二次免疫后7天(第28天)收集PBMC，并且通过ELISPOT测定测量了T细胞反应。来自用BV2373(5μg或25μg)和MATRIX-M^TM免疫的动物的PBMC具有最高数量的IFN-γ分泌性细胞，其是单独用25μg BV2373或BV2373(1μg)和MATRIX-M^TM免疫的动物的所述数量的5倍(图28)。通过ICCS分析，用BV2373(5μg)和MATRIX-M^TM免疫显示出IFN-γ+、IL-2+和TNF-α+CD4+T细胞的最高频率(图29A-图29C)。这种趋势对于同时产生至少两种或三种1型细胞因子的多功能性CD4+T细胞也是如此(图29D-图29E)。

实例4.

冠状病毒刺突(S)多肽纳米颗粒疫苗的结构表征

使用透射电子显微术(TEM)和二维(2D)类别取平均值来确定BV2373的超微结构。负染色的BV2373的高放大率(67,000倍和100,000倍)TEM图像显示出对应于S蛋白同型三聚体的颗粒。

使用自动化拣取方案构建2D类平均图像(Lander G.C.等人J Struct Biol.166,95-102(2009)；Sorzano C.O.等人,J Struct Biol.148,194-204(2004))。对同型三聚体结构进行的两轮2D类别取平均值揭示出了具有15nm长度和13nm宽度的三角形颗粒外观(图10，左上方)。将最近解析的SARS-CoV-2刺突蛋白(EMD ID：21374)的cryoEM结构覆盖在2DBV2373图像上显示出与冠形S1(NTD和RBD)和S2茎的良好拟合(图10，左下方)。在2D图像中同样明显的是从与NTD/RBD冠相对的三聚体结构的尖端突出的微弱投影(图10，右上方)。使用较大盒尺寸进行的2D类别取平均值表明，这些微弱的投影形成了S三聚体与无定形结构之间的连接。(图10，右下方)。

动态光散射(DLS)表明，与BV2365(33.4nm)和BV2373(27.2nm)的小2倍的颗粒尺寸相比，野生型CoV S蛋白具有69.53nm的Z-avg粒径。多分散性指数(PDI)表明，与野生型刺突蛋白(PDI＝0.46)相比，BV2365和BV2373颗粒在尺寸、形状和质量方面总体上是均匀的(PDI＝0.25-0.29)(表3)。

表3：SARS-CoV-2三聚体刺突蛋白的颗粒尺寸和热稳定性

通过差示扫描量热法(DSC)测定了S三聚体的热稳定性。野生型CoV S蛋白的热转变温度(T_max＝58.6℃)与分别为T_max＝61.3℃和60.4℃的BV2365和BV2373相似(表3)。更重要的是，与展开WT刺突蛋白所需的较低焓(ΔHcal＝153kJ/mol)相比，展开BV2365和BV2373变体所需的转变焓增加3-5倍(分别为ΔHcal＝466和732kJ/mol)。这些结果与和WT刺突蛋白的热稳定性相比BV2365和BV2373的热稳定性得到改善一致(表3)。

通过动态光散射评价了CoV刺突(S)多肽纳米颗粒疫苗的稳定性。使用各种pH、温度、盐浓度和蛋白酶来比较CoV刺突(S)多肽纳米颗粒疫苗与含有天然CoV刺突(S)多肽的纳米颗粒疫苗的稳定性。

实例5.

冠状病毒刺突(S)多肽纳米颗粒疫苗的稳定性

通过动态光散射评价了CoV刺突(S)多肽纳米颗粒疫苗的稳定性。使用各种pH、温度、盐浓度和蛋白酶来比较CoV刺突(S)多肽纳米颗粒疫苗与含有天然CoV刺突(S)多肽的纳米颗粒疫苗的稳定性。在不同的环境胁迫条件下使用hACE2捕获ELISA评估了没有2-脯氨酸取代的BV2365和具有两个脯氨酸取代的BV2373的稳定性。将BV2373在极端pH下(在pH 4和pH 9下48小时)、伴随长时间搅拌(48小时)和通过冷冻/解冻(2个循环)以及在升高的温度下(在25℃和37℃下48小时)孵育对hACE2受体结合没有影响(IC50＝14.0-18.3ng mL-1)。

使用过氧化氢的氧化条件将hACE2与BV2373的结合降低了8倍(IC50＝120ng mL-1)(图12A)。没有2-脯氨酸取代的BV2365不太稳定，如通过在多种条件下hACE2结合的显著损失所确定的(图12B)。

比较了BV2384(SEQ ID NO:110)和BV2373(SEQ ID NO:87)的稳定性。BV2384具有GSAS(SEQ ID NO:97)的弗林蛋白酶切割位点序列，而BV2373具有QQAQ(SEQ ID NO:7)的弗林蛋白酶切割位点。如通过SDS-PAGE和蛋白质印记所展现的，与BV2373相比，BV2384显示出广泛的降解(图32)。此外，与BV2373(图34)相比，扫描光密度测定法和回收率数据展现了全长CoV S蛋白BV2384的预料不到的损失、较低的纯度和回收率(图33)。

实例6

在食蟹猴中的免疫反应

评估了BV2373在食蟹猴SARS-CoV-2感染模型中诱导的免疫反应。第1-6组按表4所示进行治疗。

表4：食蟹猴研究的第1-6组

施用包含BV2373的疫苗导致诱导抗CoV-S抗体(图35A)，包括中和抗体(图35B)。在施用一个(图38A)或两个剂量(图38B)的BV2373后诱导了抗CoV-S抗体。施用包含BV2373的疫苗还导致产生阻断CoV S蛋白与hACE2结合的抗体(图38C和图38D)。在施用BV2373后，食蟹猴中抗CoV S多肽IgG滴度与hACE2抑制滴度之间存在显著相关性(图38E)。通过致细胞病变效应(CPE)(图40A)和噬斑减少中和测试(PRNT)(图40B)评价了BV2373诱导中和抗体产生的能力。数据揭示，与对照相比，表4的疫苗制剂产生了SARS-CoV-2中和滴度。

将包含能够在食蟹猴中诱导抗CoV-S抗体和阻断hACE2与CoV S蛋白结合的抗体的BV2373的疫苗与人恢复期血清进行了比较。数据揭示，与人恢复期血清相比，BV2373疫苗制剂诱导了优异的抗CoV S多肽和hACE2抑制滴度(图39)。

BV2373疫苗制剂还引起了SARS-CoV-2病毒复制的减少(图36A-图36B)。在用感染性病毒攻击后2天和4天(d2pi和d4pi)，在支气管灌洗液(BAL)中评估病毒RNA(图36A，对应于存在的总RNA)和病毒亚基因组RNA(sgRNA)(图36B，对应于复制病毒)水平。大多数受试者未显示出病毒RNA。在第2天，在一些受试者中测量到少量RNA。到第4天，除了接受2.5μg最低剂量的两名受试者以外，都未测量到RNA。除了同样接受最低剂量的1名受试者以外，在2天或4天均未检测到亚基因组RNA。在感染后2天和4天(d2pi和d4pi)通过鼻拭子评估病毒RNA(图37A)和病毒亚基因组(sg)RNA(图37B)。大多数受试者未显示出病毒RNA。在第2天和第4天，在一些受试者中测量到少量RNA。在2天或4天均未检测到亚基因组RNA。受试者在第0天进行免疫，并且在用两个剂量的组中在第0天和第21天进行免疫。这些数据表明，疫苗将鼻总病毒RNA降低100-1000倍，并将sgRNA降低至不可检测的水平，并且确认对疫苗的免疫反应将阻断病毒复制并防止病毒传播。

实例7

CoV S多肽纳米颗粒疫苗在人中的评价

在131名18-59岁健康参与者的随机、观察者盲性、安慰剂对照的1期临床试验中评估了包含BV2373的疫苗的安全性和功效。参与者进行了两次肌内注射免疫，间隔21天。参与者接受加或不加MATRIX-M^TM的BV2373(n＝106)或安慰剂(n＝25)。组A-E按表5所示进行治疗。图41显示了临床终点的评价的时间线。

表5：1期人类研究的组A-E

总体反应原性是轻度的，并且疫苗接种耐受性良好。在用BV2373和MATRIX-M^TM治疗的患者中，局部反应原性更为常见(图42A-图42B)。

评价了加和不加MATRIX-M^TM的BV2373的免疫原性。疫苗接种后21天，所有疫苗方案均检测到抗CoV-S抗体(图43A)。包含MATRIX-M^TM的疫苗方案的几何平均倍数上升(GMFR)超过了未佐剂化BV2373诱导的几何平均倍数上升。第二次疫苗接种后7天(第28天)，抗CoV-S滴度相对于第一次疫苗接种时观察到的反应再增加了八倍，并且在14天(第35天)内反应又再次增加超过一倍，达到GMFR相对于用单独BV2373观察到的GMFR是大约100倍。用BV2373/MATRIX-M^TM单次疫苗接种达到的抗CoV-S滴度水平与无症状(暴露)COVID-19患者中的水平相似。第二次疫苗接种达到的GMEU水平超过了门诊治疗的COVID-19患者的恢复期血清六倍，达到了与COVID-19住院治疗患者的恢复期血清相似的水平，并且超过总恢复期血清抗CoV-S抗体近六倍。两剂量5μg和25μg BV2373/MATRIX-M^TM方案的反应是相似的。这突出了佐剂(MATRIX-M^TM)使得能够节省剂量的能力。

在用BV2373治疗的所有组中均诱导了中和抗体(图43B)。用BV2373和MATRIX-M^TM方案治疗的组展现出的GMFR为单独用BV2373治疗的组的大约五倍(图43B)。与在没有佐剂的情况下进行单次疫苗接种相比，在有佐剂的情况下进行第二次疫苗接种对中和抗体滴度具有深远的影响，诱导>100倍的升高。当与恢复期血清相比时，用BV2373/MATRIX-M^TM进行第二次疫苗接种达到的GMT水平是门诊治疗的COVID-19患者的四倍，其水平跨越COVID-19住院治疗患者的水平，并且超过了总体恢复期血清GMT四倍。

从具有需要医疗护理的临床症状的COVID-19患者获得的恢复期血清展现出随着疾病严重性增加而增加的成比例的抗CoV-S IgG和中和滴度(图43A-图43B)。

在用BV2373和MATRIX-M^TM治疗的患者中观察到中和抗体滴度与抗CoV-S IgG之间的强相关性(r＝0.9466，图44C)，类似于在用恢复期血清治疗的患者中观察到的情况(r＝0.958)(图44A)。在施用未佐剂化BV2373的受试者中未观察到这种相关性(r＝0.7616)(图44B)。5μg和25μg BV2373/MATRIX-M^TM(表5的组C-E)均展现出相似的两剂量反应幅度，并且当利用两剂量方案时，每名参与者都使用任一测定测量进行了血清转化。

16名参与者(组A至D各四名参与者)的T细胞反应表明，在用BV2373刺激后，在IFN-γ、IL-2和TNF-α产生方面，BV2373/MATRIX-M^TM方案诱导了抗原特异性多功能性CD4+T细胞反应。存在对Th1细胞因子产生的强烈偏向(图45A-图45D)。

实例8

下一代CoV S多肽纳米颗粒的表达、纯化和评价

在杆状病毒表达系统中表达具有SEQ ID NO:112、SEQ ID NO:113、SEQ ID NO:114或SEQ ID NO:115的氨基酸序列的CoV S多肽，并且挑选并确认表达冠状病毒刺突(S)多肽的重组噬斑。使用具有SEQ ID NO:5的氨基酸序列的N末端信号肽来表达具有SEQ ID NO:112、SEQ ID NO:113、SEQ ID NO:114和SEQ ID NO:115的序列的CoV S多肽。

具有SEQ ID NO:112的序列的CoV S多肽包含Asn-488至酪氨酸的突变、Lys-973和Val-974至脯氨酸的突变以及具有QQAQ(SEQ ID NO:7)的氨基酸序列的灭活的弗林蛋白酶切割位点。

具有SEQ ID NO:113的序列的CoV S多肽包含Asp-601至甘氨酸的突变、Asn-488至酪氨酸的突变、Lys-973和Val-974至脯氨酸的突变以及具有QQAQ(SEQ ID NO:7)的氨基酸序列的灭活的弗林蛋白酶切割位点。

具有SEQ ID NO:114的序列的CoV S多肽包含氨基酸56、57和131的缺失、Asn-488至酪氨酸的突变、Ala-557至天门冬氨酸的突变、Asp-601至甘氨酸的突变、Pro-668至组氨酸的突变、Thr-703至异亮氨酸的突变、Ser-969至丙氨酸的突变、Asp-1105至组氨酸的突变、Lys-973和Val-974至脯氨酸的突变以及具有QQAQ(SEQ ID NO:7)的氨基酸序列的灭活的弗林蛋白酶切割位点。

具有SEQ ID NO:115的序列的CoV S多肽包含Asn-488至酪氨酸的突变、Asp-67至丙氨酸的突变、Leu-229至组氨酸的突变、Asp-202至甘氨酸的突变、Lys-404至天门冬酰胺的突变、Glu-471至赖氨酸的突变、Ala-688至缬氨酸的突变、Asp-601至甘氨酸的突变、Lys-973和Val-974至脯氨酸的突变以及具有QQAQ的氨基酸序列的灭活的弗林蛋白酶切割位点。

如实例1中那样产生CoV S多肽纳米颗粒。如实例2-7中那样评价具有SEQ ID NO:112、SEQ ID NO:112、SEQ ID NO:113和SEQ ID NO:115的氨基酸序列的CoV S多肽的稳定性和免疫原性。

实例9

BV2373和皂苷佐剂诱导针对异源SARS-CoV-2毒株的保护性免疫反应

目的：我们在英国33个地点在接受了两次间隔21天的肌内5μg剂量的BV2373和皂苷佐剂(级分A和级分C iscom基质，在本实例中也称为MATRIX-M^TM)或安慰剂(1:1)的18-84岁成人中进行了一项3期、随机、观察者设盲、安慰剂对照试验。主要功效终点是在第二次疫苗接种后7天发作的病毒学证实的轻度、中度或重度COVID-19。

共有15,187名参与者被随机化，其中7569名参与者接受了BV2373和MATRIX-M^TM，并且7570名参与者接受了安慰剂；27.8％为65岁或以上，并且4％具有SARS-CoV-2感染的基线血清学证据。BV2373和MATRIX-M^TM接受者中有10例COVID-19，并且安慰剂接受者中有96例，且在第二次疫苗接种后至少7天症状发作；BV2373和MATRIX-M^TM在预防COVID-19方面的有效性为89.7％(95％置信区间，80.2至94.6)。有五例重度COVID-19，所有病例均在安慰剂组中报告。事后分析揭示，针对原型SARS-CoV-2毒株和B.1.1.7变体的功效分别为96.4％(73.8至99.5)和86.3％(71.3至93.5)。原型SARS-CoV-2毒株包含具有SEQ ID NO:2的氨基酸序列的CoV S蛋白。B.1.1.7变体包含具有氨基酸56、57和131的缺失以及N488Y、A557D、D601G、P668H、T703I、S969A和D1105H的突变的CoV S蛋白，其中CoV S多肽关于具有SEQ ID NO:2的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。在包括具有合并症的参与者和≥65岁的参与者的各亚组之间的疫苗功效是相似的。反应原性通常是轻度和短暂的，并且在施用了BV2373和MATRIX-M^TM的组中更频繁地发生。严重不良事件发生率在两组中较低且相似。BV2373和MATRIX-M^TM的两剂量方案对原型和B.1.1.7变体的掺混物具有89.7％功效，且安全性特征与其他授权的COVID-19疫苗的安全性特征相似。

方法：试验设计和参与者：我们评估了间隔21天肌内施用的两个5μg剂量的BV2373和MATRIX-M^TM或安慰剂的安全性和有效性。这项3期试验在英国的33个招募地点进行。符合条件的参与者是18至84岁(含端值)的男性和非妊娠女性，他们是健康的或患有稳定的慢性医学病症，包括但不限于人类免疫缺陷病毒以及心脏和呼吸系统疾病。筛查时评估的健康状况是基于病史、生命体征和体检。关键排除标准包括有记录的COVID-19病史、使用免疫抑制疗法的治疗或诊断为患有免疫缺陷病。

根据预先生成的随机化时间表，使用集中的交互式响应技术系统将参与者经由区组随机化以1:1比率随机分配以接受间隔21天的两个剂量的BV2373和MATRIX-M^TM或安慰剂(生理盐水)。随机化按地点和年龄≥65岁进行分层。在一项400人的子研究中，参与者接受与所述第一个剂量一起的伴随剂量的季节性流感疫苗。这是一项观察者设盲的研究。

每次接种疫苗后，参与者在研究地点保持观察至少30分钟以监测是否存在任何急性反应。在参与者的亚组(征集性不良事件亚组)中，在每个剂量后的7天内经由电子日志收集征集性局部和全身性不良事件。评估了所有参与者从第一个剂量直到第二个剂量后28天的非征集性不良事件；评估了从第一个剂量直到第二个剂量后1年的严重不良事件、特别关注的不良事件和医疗护理不良事件。报告了接受至少一个剂量的疫苗或安慰剂的所有参与者的安全性数据。

安全性和功效：主要终点是BV2373和MATRIX-M^TM针对首次出现的病毒学证实的在基线时血清阴性的参与者中第二次疫苗接种后至少7天发作的症状性轻度、中度或重度COVID-19的功效。症状性COVID-19是根据美国食品和药物管理局(FDA)标准定义的。

在整个试验过程中监测疑似COVID-19的症状，并使用COVID-19电子症状日志(流感患者报告的结局问卷)收集其至少10天。在COVID-19的疑似症状发作时，在3天的时间内每天收集鼻子和咽喉的呼吸道样本以确认SARS-CoV-2感染。使用聚合酶链式反应(PCR)测试(UK DHSC实验室)与Thermo TaqPath^TM系统(Thermo Fisher Scientific，沃尔瑟姆，马萨诸塞州，美国)进行病毒学证实。

在接受至少一个剂量的BV2373和MATRIX-M^TM或安慰剂的所有参与者中分析安全性并进行描述性总结。还通过FDA毒性分级标准和每次注射后的持续时间总结了征集性局部和全身性不良事件。非征集性不良事件使用监管活动医学词典(MedDRA)23.1版按首选术语和系统器官类别进行编码，并按严重程度和与研究疫苗的关系进行总结。

依据对于实现主要终点的统计显著性所预期的事件总数设计并驱动试验—100例轻度、中度或重度Covid-19病例的目标。选择用于最终分析的100例目标数量以提供>95％把握度(power)以实现70％或更高的疫苗功效。基于使用Pocock边界条件的总预期主要终点的约50％(50个事件)的累积，对功效进行了单次中期分析。在对于主要终点的总体单侧I型错误率0.025下进行了对于主要目标的中期和最终分析的主要(假设检定)事件驱动的分析。在基线时呈血清阴性、接受了两个剂量的研究疫苗或安慰剂、没有影响主要终点的重大方案偏差，并且在第二次注射后6天内没有确认的症状性Covid-19病例的参与者(符合方案功效群体)中分析了主要终点。疫苗功效定义为E(％)＝(1-RR)×100，其中RR＝两个研究组(BV2373和MATRIX-M^TM或安慰剂)之间的发病率的相对风险。平均疾病发病率被报告为1000人中每年的发病率。估计的RR及其置信区间(CI)是使用泊松回归在稳健误差方差下得出的。针对无效假设：H0：疫苗功效≤30％进行了主要终点的假设检定。成功标准需要拒绝无效假设以证明统计上显著的疫苗功效。

在2020年9月28日至11月28日之间，筛选了16,645名参与者，并将15,187名参与者随机化(图47)。共有15,139名参与者接受了至少一个剂量的BV2373和Matrix-M^TM(7569)或安慰剂(7570)，其中14,039名参与者(BV2373和MATRIX-M^TM组中的7020和安慰剂组中的7019)满足对于符合方案功效群体的标准。在符合方案功效群体中基线人口统计学在BV2373和MATRIX-M^TM与安慰剂组之间平衡良好，其中48.4％为女性，94.5％为白人，0.4％为黑人或非裔美国人，0.8％为西班牙人或拉丁美洲人，并且44.6％具有至少一种伴随疾病病症(基于疾病控制和预防中心[CDC]定义)。这些参与者的中值年龄为56岁，并且27.9％为≥65岁。表6提供了临床试验参与者的基线人口统计学的总结。

表6：临床试验参与者的人口统计学和基线特征

SD，标准偏差；体重指数(BMI)计算为重量(kg)除以身高(m)的平方。百分比是基于每次治疗内和总体上设定的符合方案功效分析。*共病受试者是那些被鉴定的具有被报告为病史的共病病症的至少一种或具有筛查BMI值大于30kg/m²的人。

征集性不良事件亚组包括2714名参与者。总体而言，BV2373和MATRIX-M^TM接受者在第一个剂量(59.4％相比于20.9％)和第二个剂量(80.2％相比于17.0％)两个剂量之后报告征集性局部不良事件的频率均高于安慰剂接受者(图50)。

在BV2373和MATRIX-M^TM接受者中，在第一个剂量(54.9％和30.7％)和第二个剂量(76.6％和51.9％)两个剂量之后最常报告的局部不良事件均为注射部位压痛和疼痛，其中大多数事件在严重程度方面为1级(轻度)或2级(中度)，并且平均持续时间较短(在第一个剂量后2.3天和1.7天，并且在第二个剂量后2.8天和2.2天)。较年轻的BV2373和MATRIX-M^TM接受者(18至64岁)比较年长的BV2373和MATRIX-M^TM接受者(≥65岁)更频繁地报告征集性局部不良事件。

总体而言，BV2373和MATRIX-M^TM接受者在第一个剂量(47.6％相比于37.9％)和第二个剂量(64.6％相比于30.8％)两个剂量之后报告征集性全身性不良事件的频率均高于安慰剂接受者(图50)。在BV2373和MATRIX-M^TM接受者中，在第一个剂量(24.5％、22.3％和20.5％)和第二个剂量(40.7％、41.1％和41.0％)两个剂量之后最常报告的全身性不良事件均为头痛、肌肉疼痛和疲劳，其中大多数事件在严重程度方面为1级或2级，并且平均持续时间较短(在第一个剂量后1.6天、1.5天和1.9天，并且在第二个剂量后1.9天、1.8天和1.9天)。在第一个剂量后在两名BV2373和MATRIX-M^TM参与者中报告了4级全身性不良事件，并且在第二个剂量后在一名BV2373和MATRIX-M^TM参与者中报告了4级全身性不良事件。较年轻的疫苗接种者比较年长的疫苗接种者更频繁地报告全身性不良事件，并且在剂量2后比在剂量1后更频繁地报告全身性不良事件。值得注意的是，在第一个剂量和第二个剂量后在2.3％和5.1％的BV2373和MATRIX-M^TM参与者中报告了发热(体温≥38℃)，其中在第一个剂量和第二个剂量后分别在0.4％和0.6％的参与者中报告了3级发热(39℃-40℃)；在每个剂量的疫苗后均报告了一次4级发热(>40℃)。

截至最终功效分析的数据截止日期接受了至少一个剂量的疫苗或安慰剂的所有15,139名参与者均针对非征集性不良事件进行评估。在BV2373和MATRIX-M^TM接受者中未记录设定的不良事件的频率高于安慰剂接受者(25.3％相比于20.5％)，其中重度不良事件(1.0％相比于0.8％)、严重不良事件(0.5％相比于0.5％)、医疗护理不良事件(3.8％相比于3.9％)、导致疫苗中断的不良事件(0.3％相比于0.3％)或导致研究中断的不良事件(0.2％相比于0.2％)、潜在的免疫介导的医学病症(<0.1相比于<0.1％)以及与COVID-19相关的特别关注的不良事件(0.1％相比于0.3％)具有相似频率。在BV2373和MATRIX-M^TM接受者中报告了一个相关的严重不良事件(心肌炎)，这被认为是一种潜在的免疫介导的病症；独立的SMC认为所述事件很可能是病毒性心肌炎。参与者得到恢复。没有过敏性反应发作，并且也没有疫苗相关的疾病加重的迹象。报告了两例COVID-19相关的死亡，一例在BV2373和MATRIX-M^TM组中且在接受单个疫苗剂量后7天症状发作，并且另一例在安慰剂组中。

在符合方案功效群体中的14,039名参与者中，有10例病毒学证实的、症状性轻度、中度或重度COVID-19且在疫苗接受者中第二个剂量后至少7天发作(每1000人年有6.53人；95％CI：3.32至12.85)以及在安慰剂接受者中为96例(每1000人年有63.43人；95％CI：45.19至89.03)，疫苗功效为89.7％(95％CI，80.2至94.6；图49)。在≥65岁的患有轻度、中度或重度Covid-19的10例病例中，一例已经接受过BV2373和MATRIX-M^TM并且九例已经接受过安慰剂。在五名参与者中发生了重度COVID-19，其中没有人接受过BV2373和MATRIX-M^TM，并且五名已经接受过安慰剂。符合方案疫苗接种者中没有住院治疗或死亡。≥65岁参与者中的疫苗功效为88.9％(95％CI，12.8至98.6)，并且在剂量1之后14天的功效为83.4％(95％CI，73.6至89.5)(图49)。对主要终点的事后分析鉴定了29、66和11例Covid-19，其中分离毒株分别为SARS CoV-2原型毒株、SARS-CoV-2B.1.1.7变体或未知毒株。未知样品是那些使用非DHSC PCR测试进行PCR测试(例如，在当地医院实验室)的样品，其中未进行变体测定。针对原型毒株的疫苗功效为96.4％(95％CI，73.8至99.4)，而针对B.1.1.7变体的功效为86.3％(95％CI，71.3至93.5)。(图49)。

讨论：发现间隔21天给予的BV2373和MATRIX-M^TM的两剂量方案是安全的，并且针对由原型和B.1.1.7变体二者引起的症状性COVID-19的有效性为89.7％。本研究中累积病例的时间允许针对包括B.1.1.7变体在内的不同毒株的疫苗功效进行事后评估，所述变体现在在英国以外广泛传播，并且预计很快将在美国成为最主要的毒株。已知这种变体比先前的毒株更具传染性，并且与更高的病死率相关，这强调了对有效疫苗的需求。这是第一个在3期试验中证明对B.1.1.7变体具有高疫苗功效(86.3％)的疫苗。尽管所述研究没有把握评估对于单独SARS-CoV-2毒株的功效，但BV2373和皂苷佐剂对在试验参与者中检测到的所有毒株均显示出显著功效。特别地，针对原型毒株测定的96.4％功效点估计值与对于BNT161b2 mRNA疫苗(95.0％)和mRNA-1273疫苗(94.1％)报告的针对该毒株的功效相似，并且高于腺病毒载体疫苗所显示的功效。

最后，BV2373和皂苷佐剂组合物也显示出针对B.1.351变体的功效。

预防重度疾病(包括住院治疗、重症监护和死亡)是疫苗接种计划的重要目标，并且BV2373和皂苷佐剂的两剂量方案显示出非常高的功效，类似于对于其他获得许可的Covid-19疫苗所报告的功效。另外，BV2373和皂苷佐剂在第一个剂量后提供的保护水平与其他COVID-19疫苗的保护水平在相似范围内。在BV2373和皂苷佐剂的1/2期研究期间观察到的有利的安全性特征在该3期试验中得到证实。反应原性一般为轻度或中度，并且在老年受试者中反应不太常见且较轻，并且在第二个剂量后更常见。注射部位压痛和疼痛、疲劳、头痛和肌肉疼痛是最常报告的局部和全身性不良事件，并且在使用疫苗的情况下比安慰剂更常见。疫苗组和安慰剂组中的严重不良事件发生率相似(各为0.5％)，并且没有因接种疫苗而导致的死亡。

该试验的结果提供了进一步的证据证明由原型SARS-CoV-2和SARS-CoV-2变体B.1.1.7引起的COVID-19可以通过免疫来预防，为基于蛋白质的佐剂化疫苗提供了第一个证据。这些数据证实BV2373和皂苷佐剂可以在标准冰箱温度下储存，此外，可以诱导对刺突蛋白抗原的广泛表位反应。这种广泛的反应提供了对一系列异源SARS-CoV-2毒株的保护性功效。

实例10

BV2438和皂苷佐剂诱导针对异源SARS-CoV-2毒株的保护性免疫反应

目的：评价了与皂苷佐剂的组合的含有重组CoV刺突(rS)蛋白BV2438(SEQ ID NO:132)、BV2373(SEQ ID NO:87)或两者的组合物的免疫原性和体内保护。所述皂苷佐剂含有两种iscom颗粒，其中：第一iscom颗粒包含皂树的级分A而不包含皂树的级分C；并且第二iscom颗粒包含皂树的级分C而不包含皂树的级分A。级分A和级分C分别占在所述佐剂中皂树的级分A和皂树的级分C的重量之和的按重量计85％和15％。

评价了单独或与前述皂苷佐剂组合的BV2438和BV2373免疫方案针对SARS-CoV-2/WA1、SARS-CoV-2/B.1.1.7和SARS-CoV-2/B.1.351毒株的功效。SARS-CoV-2/WA1毒株具有含有SEQ ID NO:2的氨基酸序列的CoV S多肽。SARS-CoV-2/B.1.1.7毒株具有包含氨基酸69、70和144的缺失以及N501Y、A570D、D614G、P681H、T716I、S982A和D1118H的突变的CoV S多肽，其中所述CoV S多肽关于具有SEQ ID NO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。SARS-CoV-2/B.1.351毒株具有包含D80A、L242H、R246I、A701V、N501Y、K417N、E484K和D614G的突变的CoV S多肽，其中所述CoV S多肽关于具有SEQ ID NO:1的氨基酸序列的野生型SARS-CoV-2S多肽进行编号。

方法：

细胞和病毒：病毒和细胞如前所述进行处理(18)。简而言之，将Vero E6细胞(ATCC#CRL 1586)在补充有10％(v/v)胎牛血清(Gibco)、1％(v/v)青霉素/链霉素(GeminiBio-产品)和1％(v/v)L-谷氨酰胺(2mM终浓度，Gibco)的DMEM(Quality Biological)(Vero培养基)中培养。将细胞在37℃和5％CO2下维持。SARS-CoV-2/WA1由CDC(BEI#NR-52281)提供。SARS-CoV-2/B.1.17和SARS-CoV-2/B.1.351由约翰霍普金斯大学的Andy Pekosz博士慷慨提供。当CPE开始可见时，通过感染Vero E6细胞两天来制备两种病毒的原液。通过离心收集和澄清培养基然后进行等分以在-80℃下储存。如前所述，使用Vero E6细胞通过噬斑测定检测原液的滴度。

SARS-CoV-2蛋白表达：SARS-CoV-2构建体由全长S糖蛋白基因序列(GenBankMN908947核苷酸21563-25384)合成产生。全长S基因经过密码子优化以在草地贪夜蛾(Sf9)细胞中表达，并由GenScript(Piscataway，新泽西州，美国)合成产生。QuikChangeLightning定点诱变试剂盒(Agilent)用于产生两种刺突蛋白变体：弗林蛋白酶切割位点(682-RRAR-685)突变为682-QQAQ-685以具有蛋白酶抗性，并且在位置K986P和V987P(2P)处的两个脯氨酸取代被引入以产生双突变体BV2373。为了生成基于B.1.351变体的重组刺突构建体，还引入了以下点突变：D60A、D215G、L242H、K417N、E484K、N501Y、D614G和A701V。在苜蓿银纹夜蛾多角体启动子的转录控制下，在pFastBac杆状病毒转移载体(Invitrogen，卡尔斯巴德，加利福尼亚州)中的BamHI-HindIII位点之间选殖了全长S基因。重组杆状病毒构建体经噬斑纯化，并且制备主种子原液并将其用于产生工作病毒原液。使用快速滴度试剂盒(Clontech，山景城，加利福尼亚州)测定杆状病毒主原液滴度和工作原液滴度。通过以≤0.01噬斑形成单位(pfu)/细胞的感染复数(MOI)感染Sf9细胞来制备重组杆状病毒原液。

表达和纯化：如前所述，在Sf9细胞中产生SARS-CoV-2S蛋白。简而言之，将细胞在无血清培养基中扩增并用重组杆状病毒感染。将细胞在27℃±2℃下培养，并在感染后68-72小时通过离心(4000×g 15min)收获。将细胞沈淀物悬浮于含有亮抑酶肽的25mM TrisHCl(pH 8.0)、50mM NaCl和0.5％-1.0％(v/v)聚氧乙烯壬基苯酚(NP-9，)NP-9中。用含有NP-9洗涤剂的Tris缓冲液从质膜提取S蛋白，通过以10,000x g离心30min澄清。通过TMAE阴离子交换和小扁豆凝集素亲和层析纯化S蛋白。中空纤维切向流过滤用于在25mM磷酸钠(pH 7.2)、300mM NaCl、0.02％(v/v)聚山梨醇酯80(PS 80)中配制100-150μgmL-1的纯化刺突蛋白。通过用Gel-Code Blue试剂(Pierce，罗克福德，伊利诺伊州)染色的4％-12％梯度SDS-PAGE评价纯化的S蛋白，并使用OneDscan系统(BD Biosciences，罗克维尔，马里兰州)通过扫描光密度测定法测定纯度。

差示扫描量热法：样品(分别为：BV2426 Lot 01Feb21和BV2373 Lot 15Dec20；rS-B.1.351BV2438和rS-WU1BV2373)和相应的缓冲液以每分钟1℃的速度从4℃加热到120℃，并且在NanoDSC(TA Instruments，纽卡斯尔，特拉华州)中测量差热容量变化。进行单独的缓冲液扫描以获得基线，从样品扫描中减去基线以产生基线校正的曲线。峰顶点所在的温度是转变温度(Tmax)，并且峰下面积提供转变焓(ΔHcal)。

透射电子显微术和2D类平均：电子显微术由NanoImaging Services(圣地亚哥，加利福尼亚州)使用配备有FEI Eagle 4k x 4k CCD相机的在120keV下运行的FEI TecaniT12电子显微镜进行。将SARS-CoV-2S蛋白在制剂缓冲液中稀释至2.5μg mL-1。将样品(3μL)施加至硝酸纤维素支撑的400目铜网格，并用铀酰形式染色。在多个尺度上获取每个网格的图像以评估样品的整体分布。以150,000倍(X nm/像素)和92,000倍(0.16nm/像素)的标称放大倍率获取高放大倍率图像。这些图像是在-2.0μm至-1.5μm(110,000倍)的标称散焦和约的电子剂量下获取的。

对于类平均，从92,000倍高放大倍率图像识别出颗粒，然后如前所述进行比对和分类。

通过生物发光成像(BLI)测定SARS-CoV-2S与hACE2受体结合的动力学：使用OctetQK384系统(Pall FortéBio，菲蒙市，加利福尼亚州)通过生物膜层干涉测量术(BLI)测定S蛋白受体结合动力学。将His标记的人ACE2(2μg mL-1)固定在带镍的Ni-NTA生物传感器尖端上。在基线后，SARS-CoV-2rS蛋白溶液在动力学缓冲液中在300nM至4.7nM的范围内连续稀释2倍，允许缔合600sec，然后再解离600-900sec。使用Octet软件HT 10.0通过1:1全局曲线拟合分析数据。

小鼠研究设计：雌性BALB/c小鼠(7-9周龄，17-22克，每组N＝20只)通过肌内(IM)注射进行免疫，其中单独的、组合的或作为异源初次免疫/加强免疫的与5μg基于皂苷的Matrix-M^TM佐剂(Novavax,AB，乌普萨拉，瑞典)一起的rS-WU1BV2373、rS-B.1.351BV2438的两个剂量间隔14天(研究第0天和第14天)。安慰剂组被注射作为阴性对照的疫苗制剂缓冲液。在研究第-1天、第14天、第21天和第32天收集血清进行分析。接种疫苗的动物和对照动物在研究第46天用SARS-CoV-2进行鼻内攻击。

为了评估基质-皂苷佐剂介导的细胞反应，用上述相同的方案对雌性BALB/c小鼠组(每组N＝8只)进行IM免疫，其中注射间隔21天。在第二次免疫后7天(研究第28天)收集脾脏。未接种疫苗的组(N＝5)用作对照。

狒狒研究设计：将九只成年狒狒(研究开始时为10-16岁)随机分为4组，每组2-3只，并通过IM注射1、5或25μg rS下的rS-WU1BV2373与50μg基质-皂苷佐剂进行免疫。一个单独的组用25μg rS在无佐剂的情况下进行免疫。在该初次免疫系列中对动物进行疫苗接种，且2个剂量间隔21天。初次免疫系列后的免疫原性结果如前所述(15)。大约一年(45周)后，所有动物都用一个或两个3μg剂量的rS-B.1.351BV2438与50μg基质-皂苷佐剂加强免疫。在加强免疫之前和之后收集血清和PBMC以测量抗体和细胞介导的免疫反应。

小鼠中的SARS-CoV-2攻击：通过腹膜内注射50μL在磷酸盐缓冲盐水(PBS)中的甲苯噻嗪(0.38mg/小鼠)和氯胺酮(1.3mg/小鼠)的混合物来麻醉小鼠。小鼠鼻内接种在50μL中的7×104pfu的B.1.117或1×105pfu的B.1.351SARS-CoV-2毒株。在感染当天称量受攻击的小鼠，并且在感染后每天称重持续4天。在感染后第2天和第4天，从每个疫苗接种组和对照组处死5只小鼠，并收获肺以通过噬斑测定测定病毒滴度，并通过qRT-PCR测定病毒RNA水平。

SARS-CoV-2斑块测定：通过使用1.0mm玻璃珠(Sigma Aldrich)和Beadruptor(Omini International Inc.)在PBS(Quality Biological Inc.)中使收获的肺匀浆化来量化SARS-CoV-2肺滴度。将匀浆添加到Vero E6近汇合培养物中，并且使用6点稀释曲线通过计数噬斑形成单位(pfu)来测定SARS-CoV-2病毒滴度。

通过ELISA测定抗SARS-CoV-2刺突IgG：使用ELISA测定抗SARS-CoV-2S IgG滴度。简而言之，用1.0μg mL-1的SARS-CoV-2刺突蛋白包被96孔微量滴定板(ThermoFischerScientific，罗契斯特市，纽约州，美国)。用PBS-T洗涤板并将其用TBS Startblock封闭缓冲液(ThermoFisher,Scientific)封闭。将小鼠、狒狒或人血清样品连续稀释(10-2至10-8)并添加至封闭板，然后在室温下孵育2小时。孵育后，用PBS-T洗涤板并添加HRP缀合的山羊抗小鼠IgG或山羊抗人IgG(Southern Biotech，伯明翰，亚拉巴马州，美国)持续1小时。用PBS-T洗涤板并添加3,3',5,5'-四甲基联苯胺过氧化物酶底物(TMB，T0440-IL，Sigma，圣路易斯，密苏里州，美国)。用TMB终止溶液(ScyTek Laboratories,Inc.，洛根，犹他州)终止反应。用SpectraMax Plus板读取器(Molecular Devices，美国加利福尼亚州桑尼维尔)在450nm的OD下读取板，并且用SoftMax软件分析数据。使用SoftMax Pro 6.5.1 GxP软件通过4参数拟合来计算EC50值。使用GraphPad Prism 7.05软件绘制单独动物抗SARS-CoV-2 SIgG滴度和组几何平均滴度(GMT)和95％置信区间(±95％CI)。

hACE2受体阻断抗体：通过ELISA测定人ACE2受体阻断抗体。将九十六孔板在4℃下用1.0μg mL-1 SARS-CoV-2 S蛋白包被过夜。在用PBS-T洗涤并用StartingBlock(TBS)封闭缓冲液(ThermoFisher Scientific)封闭后，将来自免疫的小鼠、狒狒或人组的连续稀释血清添加至包被的孔中，并在室温下孵育1小时。洗涤后，在室温下将30ng mL-1组氨酸标记的hACE2(Sino Biologics，北京，中国)添加至孔中持续1小时。洗涤后，添加HRP缀合的抗组氨酸IgG(Southern Biotech,Birmingham，伯明翰，亚拉巴马州，美国)，然后洗涤并添加TMB底物。用SpectraMax Plus板读取器(Molecular Devices，桑尼维尔，加利福尼亚州，美国)在450nm的OD下读取板，并且用SoftMax Pro 6.5.1GxP软件分析数据。使用SoftMax Pro程序中的以下方程序计算每个样品的每个稀释度的抑制％：100-[(平均结果/阳性对照下的对照值)*100]。

生成血清稀释度与抑制％的关系图，并通过4参数逻辑(4PL)曲线拟合至数据进行曲线拟合。在SoftMax Pro程序中测定了hACE2与SARS-CoV-2S蛋白(BV2373或BV2438)的50％抑制(IC50)时的血清抗体滴度。

通过噬斑减少中和滴度测定(PRNT)测定的SARS-CoV-2中和滴度：如前所述处理PRNT(20)。简而言之，将血清样品在DMEM(Quality Biological)中以1:40的初始稀释度和1:2的连续稀释度稀释，总共11次稀释。每个板上都包括无血清对照。然后将SARS-CoV-2以1:1添加至每个稀释液中，以达到每个噬斑测定孔50PFU的目标，并在37℃(5.0％CO2)下孵育1h。然后通过噬斑测定来测定样品滴度，并与未处理对照相比确定中和滴度。在PRISM(GraphPad，圣地亚哥，加利福尼亚州)中将4参数逻辑曲线拟合至这些中和数据，并基于该曲线拟合计算中和50％病毒所需的稀释度(PRNT50)。

表面和细胞内细胞因子染色：对于表面染色，首先将鼠脾细胞与抗CD16/32抗体一起孵育以阻断Fc受体。为了表征滤泡辅助性T细胞(Tfh)，将脾细胞与以下抗体或染料一起孵育：BV650缀合的抗CD3、APC-H7缀合的抗CD4、FITC缀合的抗CD8、Percp-cy5.5缀合的抗CXCR5、APC缀合的抗PD-1、Alexa Fluor 700缀合的抗CD19、PE缀合的抗CD49b(BDBiosciences，圣何塞，加利福尼亚州)和黄色LIVE/染料(Life Technologies，纽约州)。为了染色生发中心(GC)B细胞，将脾细胞用FITC缀合的抗CD3、PerCP-Cy5.5缀合的抗B220、APC缀合的抗CD19、PE-cy7缀合的抗CD95和BV421缀合的抗GL7(BD Biosciences)和黄色活性染料(LIVE/)(Life Technologies，纽约州)进行标记。

对于鼠脾细胞的细胞内细胞因子染色(ICCS)，将细胞以每孔2×106个细胞在96孔U型底板中培养。用rS-WU1BV2373或rS-B.1.351BV2438刺突蛋白刺激细胞。将板在37℃下孵育6h，最后4h在BD GolgiPlug^TM和BD GolgiStop^TM(BD Biosciences)的存在下孵育。用针对CD3(BV650)、CD4(APC-H7)、CD8(FITC)、CD44(Alexa Fluor 700)和CD62L(PE)(BDPharmingen，加利福尼亚州)的鼠抗体和黄色LIVE/染料标记细胞。用Cytofix/Cytoperm(BD Biosciences)固定后，将细胞与PerCP-Cy5.5缀合的抗IFN-γ、BV421缀合的抗IL-2、PE-cy7缀合的抗TNF-α和APC缀合的抗IL-4(BD Biosciences)一起孵育。使用LSR-Fortessa或FACSymphony流式细胞仪(Becton Dickinson，圣地亚哥，加利福尼亚州)获取所有染色的样品，并使用FlowJo软件Xv10版(Tree Star Inc.，阿什兰，俄勒冈)分析数据。

对于狒狒PBMC的ICS，在图5A中列出的时间点收集的PBMC如上所述用rS-WU1BV2373或rS-B.1.351BV2438进行刺激。用人/NHP抗体BV650缀合的抗CD3、APC-H7缀合的抗CD4、FITC缀合的抗CD8、BV421缀合的抗IL-2、PerCP-Cy5.5缀合的抗IFN-γ、PE-cy7缀合的抗TNF-α、APC缀合的抗IL-15、BV711缀合的抗IL-13(BD Biosciences)和黄色LIVE/活性染料标记细胞。

酶联免疫吸附测定(ELISA)：按照制造商对于小鼠IFN-γ和IL-5ELISpot试剂盒(3321-2H和3321-2A，Mabtech，辛辛那提，俄亥俄州)的程序进行鼠IFN-γ和IL-5ELISpot测定。简而言之，在用抗IFN-γ或抗IL-5抗体预包被的板中用rS-WU1BV2373或rS-B.1.351BV2438刺激200μL体积中的4×10⁵个脾细胞。检测二抗为克隆RS-6A2 IFN-γ和克隆TRFK4。每个刺激条件一式三份进行。将测定板在37℃下在5％CO2培养箱中培养24-48h，并使用BD ELISpot AEC底物组(BD Biosciences，圣地亚哥，加利福尼亚州)进行显影。使用ELISpot读取器和ImmunoSpot软件v6(Cellular Technology,Ltd.，谢克海茨，俄亥俄州)对斑点进行计数和分析。通过减去培养基对照中的背景数量获得IFN-γ或IL-5分泌性细胞的数量。图中显示的数据是一式三份孔的平均值。

类似地，使用来自Mabtech的NHP IFN-γ和人IL-4测定试剂盒进行狒狒IFN-γ和IL-4测定。对于IFN-γ，使用包被抗体人IFN-γ3420-2H和检测抗体克隆7-B6-1。对于IL-4，使用包被抗体人IL-43410-2H(克隆IL4-I)和检测抗体克隆IL4-II。一式三份地进行测定。

统计分析：使用GraphPad Prism 8.0软件(拉荷亚，加利福尼亚州)进行统计分析。如图所示绘制单独动物的血清抗体滴度和几何平均滴度(GMT)和95％置信区间(95％CI)或平均值±SEM。对log10转化数据进行普通单因素方差分析与Tukey多重比较事后检定，以评价组间差异的统计显著性。P值≤0.05被认为是统计上显著的。

BV2438抗原的生物物理特性、结构和功能：纯化的BV2438在还原并进行SDS-PAGE时迁移，且预期分子量为约170kDa(图52A)。通过差示扫描量热法(DSC)比较了BV2438与BV2373的热稳定性；与原型BV2373蛋白相比，BV2438的主峰显示热转变温度(T_max)增加4℃，且转变焓(ΔHCal)为1.3倍高，表明BV2438具有增加的稳定性(图52B，表7)。对16,049个颗粒的透射电子显微术(TEM)结合两轮二维(2D)类平均用于确认BV2438的超威结构。高放大倍率(92,000x和150,000x)TEM图像显示长15nm且宽11nm的灯泡形颗粒外观，这与SARS-CoV-2刺突三聚体的预融合形式一致(PDB ID 6VXX；图52C)。这与我们先前对于原型BV2373蛋白所观察到的一致。

为了确认变体刺突蛋白构建体BV2438的功能特性，如前所述使用生物膜层干涉测量术(BLI)测定该rS蛋白与hACE2受体的结合。发现BV2438与hACE2以3.94×10⁴的结合常数(Ka)紧密且稳定地结合，其表示与hACE2的缔合是与原型蛋白BV2373(Ka＝1.08×10⁴)相比的3.6倍。这两种蛋白质的解离常数基本相同(对于BV2438和BV2373分别为1.46×10^-7和1.56×10^-7)。我们还如前所述使用ELISA评估BV2438与hACE2的结合。在该测定中，BV2438在比原型构建体BV2373(EC₅₀＝9.4ng/mL)略低的浓度(EC₅₀＝8.0ng/mL)下得到了50％的hACE2饱和度，证实了与BV2373对于hACE2的亲和力相比，BV2438展现出更高的对于hACE2的亲和力(表7)。

表7：SARS-CoV-2重组刺突蛋白的热稳定性和hACE2结合

T_max，熔解温度；K_a，结合常数；K_dis，解离常数；EC₅₀，半最大结合。

在小鼠中的BV2438免疫原性：我们评估了与皂苷佐剂一起配制的BV2438和BV2373的抗体和细胞介导的免疫原性。为了评估抗体介导的免疫原性，小鼠组(n＝20)用BV2373或BV2438作为初次免疫和加强免疫二者均进行了免疫，其中用BV2373作为初次免疫以及BV2438作为加强免疫，或其中将两种疫苗组合在二价制剂中以进行初次免疫和加强免疫疫苗接种。安慰剂组接受作为阴性对照的疫苗制剂缓冲液。在单价免疫组中，在第0天和第14天肌内注射1μg的rS和5μg的皂苷佐剂。对于二价免疫，每次免疫时施用1μg的每种rS构建体(总共2μg rS)以及5μg的皂苷佐剂。研究设计示于图53中。用4种疫苗方案中的任何一种进行免疫的小鼠显示出在疫苗接种后第21天通过ELISA测定的针对B.2刺突和B.1.351刺突的抗体滴度升高。用BV2373或BV2438进行单价疫苗接种产生的抗S(WU1)IgG滴度显著低于二价疫苗接种或异源疫苗接种，但两者降低的IgG滴度均不超过2倍(图54A，图54B)。相反，与所有其他免疫方案相比，单独使用BV2373免疫导致显著更低的针对B.1.351刺突的滴度；使用单价BV2438或二价rS免疫导致抗B.1.351刺突IgG滴度在所测试的方案中最高，且在这些方案之间滴度没有显著差异(图54A，图54B)。正如预期的那样，安慰剂组中的动物展现出不可检测的抗B.2刺突和抗B.1.351刺突IgG滴度。

还评估了来自小鼠的血清抑制刺突与hACE2结合的能力。所有免疫方案均导致在第21天产生了阻断hACE2与CoV刺突多肽结合的抗体，且在任何组之间无显著差异(图54C，图54D)。然而，单独使用BV2373免疫导致显著更低的能够破坏B.1.351刺突与hACE2之间的结合的血清滴度；BV2373单独免疫组中的滴度比BV2438单独免疫组中的滴度低4.6倍(p<0.0001)，并且比接受二价rS的组中的滴度低3.1倍(p<0.0001)。

我们接下来评估了不同疫苗接种方案中的中和抗体滴度。在噬斑减少中和滴度测定(PRNT₅₀)中，使用SARS-CoV-2/WA1、SARS-CoV-2/B.1.1.7和SARS-CoV-2/B.1.351毒株评估从疫苗接种后第32天从疫苗接种动物收集的血清。来自单价BV2373组的血清对3种病毒株中的每一种都显示出相似的中和抗体滴度。与B.2毒株相比，来自用单价BV2438免疫的小鼠的血清产生了升高的对B.1.351和B.1.1.7毒株的中和抗体滴度(图54E)。与B.2毒株相比，异源疫苗组产生了类似升高的对B.1.351和B.1.17毒株的中和抗体滴度，二价BV2373/BV2438疫苗接种方案也是如此。

在BALB/c小鼠中针对SARS-CoV-2的BV2438保护：评价了如图53中所述进行疫苗接种的小鼠产生针对用B.1.1.7或B.1.351攻击的保护性免疫的能力。虽然SARS-CoV-2/Wuhan1(B.2)毒株不会在野生型小鼠中复制，但B.1.1.7和B.1.351毒株在刺突ORF中具有501Y突变，从而允许刺突蛋白与小鼠ACE2结合并进入细胞。在疫苗接种后第46天，小鼠鼻内接种7×10⁴PFU的B.1.17(每组n＝10只小鼠)或1×10⁵PFU的B.1.351(每组n＝10只小鼠)。在整个攻击后时间段每天对小鼠称重，在感染后第2天和第4天(研究第48天和第50天)，通过吸入异氟烷吸入处死每组5只小鼠。然后通过噬斑形成测定评估每只小鼠的肺的病毒载量，并通过RT-PCR评估病毒RNA。感染B.1.1.7的安慰剂BALB/c小鼠体重没有减轻，并且在感染这种SARS-CoV-2毒株的任何疫苗接种组中都没有观察到体重减轻。对于B.1.351感染的小鼠，截至用B.1.351感染之后第4天，在安慰剂接种组中观察到20％的体重减轻(图55A，图55B)。用任一方案进行疫苗接种的所有小鼠被保护免于在感染B.1.351后体重减轻，证明了该模型中保护的临床相关性。

在感染后第2天，安慰剂组中的B.1.1.7感染的小鼠展现出4×10⁴pfu/g肺，在安慰剂疫苗接种组中截至感染之后第4天降至不可检测的水平。在用任何BV2373或BV2438方案免疫后，在感染之后第2天或第4天没有可检测到的活病毒，证明在疫苗接种后病毒载量减少超过5个对数并且被保护免于感染(图55C，图55D)。在感染之后第2天，假疫苗接种组中的B.1.351感染的小鼠展现出8×10⁸pfu/g肺，截至感染之后第4天降至2×10⁵pfu/g肺。在用任何rS方案免疫后，在B.1.351感染的小鼠中在感染之后第2天或第4天没有可检测到的活病毒。这证明病毒滴度显著降低，其中在假疫苗接种的小鼠中截至感染之后第2天病毒载量降低>5个对数(图55C，图55D)。还针对在攻击后亚基因组(sgRNA)SARS-CoV-2mRNA产生评估了肺RNA。相对于各个安慰剂组中的水平，我们发现在感染两种毒株之后第2天和第4天免疫的小鼠中的肺sgRNA水平降低>99％(图55E，图55F)。

这些结果证实了与皂苷佐剂一起配制并作为单价、二价或异源方案施用的BV2373和BV2438在小鼠中赋予了针对SARS-CoV-2的两种毒株B.1.1.7和B.1.351的保护。连同体重减轻的减少、高中和抗体滴度以及消除小鼠肺中的病毒复制，我们证明了通过变体刺突靶向疫苗实现的高度保护性疫苗反应。

BV2438在小鼠中的细胞介导的免疫原性：BALB/c小鼠组(n＝8只/组)用相同的上述BV2373或BV2438方案进行免疫，但间隔21天(图56A)。向阴性对照组(n＝4)注射疫苗制剂缓冲液。在加强免疫之后7天，在研究第28天收获脾脏。收集脾细胞并进行ELISpot和细胞内细胞因子染色(ICS)以检查用BV2373或BV2438刺激后的细胞因子分泌。酶联免疫吸附测定(ELISA)显示在所有疫苗接种方案后与IL-5产生性细胞的数量相比更多的IFN-γ产生性细胞的数量，这表明Th1偏斜反应(图56B-图56D)。在用任一rS刺激后，通过ICS观察到强烈的Th1反应，如通过表达IFN-γ、IL-2或TNF-α的CD4+T细胞和表达所有3种细胞因子的多功能CD4+T细胞的存在所测量的(图56E，图57A-图57E)。也存在表达Th2细胞因子IL-4但对IL-2和TNF-α呈阴性的CD4+T细胞，但其比例低于对于Th1细胞因子所观察到的比例(图57A-图57E)。对于用BV2373或BV2438刺激后测试的任何细胞因子，在疫苗接种组之间没有观察到细胞因子阳性细胞数量的显著差异。

滤泡辅助性T细胞(CSCR5+PD-1+CD4+)倾向于代表更大百分比的CD4+T细胞，但与安慰剂组动物相比，在疫苗接种的动物中未观察到统计上显著的升高(图56F)。类似地，通过使用流式细胞术测定CD19+B细胞中GL7+CD95+细胞的百分比来评价生发中心形成，但与安慰剂组相比，在疫苗接种组中观察到更高百分比的生发中心B细胞的趋势，只有用单价BV2438方案免疫的动物显示出显著更高的比例(与安慰剂组相比，p＝0.049；图56G)。

在狒狒中用BV2373初次免疫之后一年用BV2373加强免疫诱导的记忆反应：小的狒狒群组(总共n＝9)进行了使用BV2373的初次免疫系列(1μg、5μg或25μg rS与50μg皂苷佐剂，或25μg未佐剂化rS)。大约一年后，所有动物都用一个或两个剂量的3μg BV2438与50μg皂苷佐剂加强免疫以检查产生的免疫反应(图58A)。在第一次BV2438加强免疫之后七天，最初接受佐剂化BV2373的动物展现出强烈的记忆反应，如通过高于最初在初次免疫系列期间在峰值免疫反应下所观察到的水平的抗S(WU1)IgG滴度水平所展现(图58B)。这种反应似乎并没有被第二个加强剂量的BV2438进一步加强，但本研究中使用的小样本量阻止了有意义的定量分析。在初次免疫系列期间接受未佐剂化BV2373的动物展现出对用BV2438加强免疫的较弱反应，但仍展现出升高的抗S(WU1)IgG反应。BV2438加强免疫引发了针对BV2373和BV2438的可比较的抗体滴度，其中最初接受未佐剂化BV2373的动物展现出较弱的反应(图58C，图58D)。

还在加强免疫前、以及用1或2个剂量的BV2438加强免疫后7、21、35和89天评价了能够破坏野生型CoV S蛋白(SEQ ID NO:2)或B.1.351rS与hACE2之间相互作用的血清抗体滴度。与对于抗S IgG滴度观察到的相似，在初次免疫系列期间接受佐剂化疫苗的动物在BV2438加强免疫之后7天展现出强烈的hACE2抑制性抗体反应，尽管在加强免疫前具有不可检测的滴度。与BV2438-hACE2阻断抗体的水平相比，对于BV2373-hACE2阻断抗体的滴度略高，但小样本量阻碍了有意义的定量分析。在初次免疫系列期间接受未佐剂化疫苗的动物在BV2438加强免疫后展现出较低的hACE2阻断滴度(图58E)。

通过测试血清中和WA1、B.1.351和B.1.1.7的能力，通过活病毒微中和测定分析中和抗体滴度。在BV2438加强免疫之前收集的血清对所有这些病毒具有不可检测的中和抗体水平。截至疫苗接种之后7天，检测到中和所有3种毒株的高滴度抗体，并且该抗体反应直到疫苗接种之后35天仍保持高位。在初次免疫系列中用未佐剂化BV2373免疫的动物显示出显著较低的抗体水平以及更广泛的中和滴度范围(图58F)。总之，这些数据证明甚至在初次疫苗接种系列之后1年仍具有稳健持久的抗体反应。

在狒狒中第一个BV2438加强剂量之后7天也观察到表达3种Th1细胞因子的多功能T细胞，并且这些反应在第一个加强剂量之后35天仍保持(图58G和图59A-图59G)。

通过来自BV2373疫苗接种的成人的血清中和SARS-CoV-2变体：一种含有BV2373和皂苷佐剂的疫苗目前正在全球(包括在B.1.1.7和B.1.351流行的地区)进行临床试验。我们评估了这些试验中来自个体的血清中和USA-WA1、B.1.1.7和B.1.351的能力。使用PRNT₅₀读数进行微量中和测定(图60A，图60B)。在第二个剂量的疫苗后，评估了来自临床试验参与者的三十个随机选择的血清样品。在比较WA1相比于B.1.1.7时，大多数血清样品的中和活性没有变化；只有1个样品具有针对B.1.1.7的中和抗体滴度的统计上显著的变化。WA1相比于B.1.351中和滴度显示中和滴度范围增加，其中30个样品中有五个显示中和降低，与PRNT₅₀测定中的平均值相差1个标准偏差。该数据证明与B.1.1.7相比，在一小部分接受BV2373和皂苷佐剂的疫苗接种者中，对B.1.351的中和降低。

讨论：我们已经证明，使用通过皂苷佐剂进行佐剂化的B.1.351刺突变体的全长、稳定的预融合SARS-CoV-2刺突糖蛋白疫苗可以诱导高水平的功能性免疫，并保护小鼠免于B.1.1.7和B.1.351SARS-CoV-2毒株二者的危害。用BV2438对小鼠或非人灵长类动物进行免疫诱导了抗S抗体、hACE2受体抑制性抗体和SARS-CoV-2中和抗体。另外，BV2438疫苗诱导了CD4⁺T细胞反应，诱导了生发中心形成并提供了针对B.1.351和B.1.1.7攻击的保护。

在小鼠中，用B.1.351变体定向疫苗进行疫苗接种后产生的抗体能够同等程度地抑制hACE2与变体刺突或原始刺突之间的结合，表明这种变体定向疫苗可以有效地“向后”保护免于原始SARS-CoV-2毒株的危害。

对来自我们试验的人疫苗血清的分析显示了稳健的抗体反应和最小的中和损失。我们观察到，与B.1.1.7和WA1相比，B.1.351病毒不会显著降低中和，尽管有证据表明南非的WA1试验参与者中的突破性感染。所有突破性感染均为B.1.351。因此，含有单个或多个变体rS疫苗的加强免疫疫苗接种将增加抗体水平并扩大对如本工作所示的变体的覆盖范围。

实例11

BV2373和皂苷佐剂在单次加强剂量之后诱导针对异源SARS-CoV-2毒株的保护性免疫反应

参与者：招募年龄≥18至≤84岁的健康男性和女性参与者以募入本研究中。如果参与者的体重指数为17至35kg/m²，能够在募入之前提供知情同意书，并且(对于女性参与者)同意保持不活跃的异性恋或使用经批准的避孕形式，则参与者符合条件。有严重急性呼吸综合征(SARS)病史或确诊为COVID-19、严重慢性医学病症(例如，糖尿病、充血性心力衰竭、自身免疫性病症、恶性肿瘤)或目前正在针对未确诊的疾病进行评估(这可能导致新的诊断)的参与者被排除在研究之外。怀孕或哺乳期女性也被排除在外。

随机化：患者被随机分配为五组。在五个治疗组中，一个是安慰剂对照组(A组)并且两个是活性疫苗组，其被考虑用加强免疫进行另外的疫苗接种(B组和C组)。大约6个月后，被随机化以在第0天和第21天(B组)接受两个剂量的BV2373(5μg)和皂苷佐剂(50μg)或在第0天接受一个剂量的BV2373(5μg)和皂苷佐剂(50μg)且在第21天接受安慰剂(C组)的同意的参与者以1:1重新随机化以在第189天接受单次加强剂量的BV2373和相同剂量水平的皂苷佐剂(B2组和C2组)或安慰剂(B1组或C1组)。B组参与者是本实例的主要焦点。

目的和方法：我们在18至84岁的健康成人中进行了一项2期、随机、观察者设盲、安慰剂对照试验，这些成人接受了三次肌内注射5μg剂量的BV2373和50μg皂苷佐剂(级分A和级分C iscom基质，在本实例中也称为MATRIX-M^TM)或安慰剂(1:1)。第一个剂量和第二个剂量间隔21天施用。第一个剂量和第二个剂量被称为“初次疫苗接种系列”。第三个剂量(“加强”剂量)在初次疫苗接种系列之后约6个月施用。所有三个剂量的注射体积均为0.5mL。评估了安全性和免疫原性参数，包括针对原始SARS-CoV-2毒株和选择变体(B.1.351[β]、B.1.1.7[α]、B.1.617.2[δ])的IgG、MN₅₀和hACE2抑制的测定。

参与者使用电子日志记录疫苗接种当天的反应原性，之后再记录6天。在接受加强免疫之后28天收集血液样品以用于免疫原性分析，此时还进行了安全性随访。免疫反应的测量包括对于血清免疫球蛋白G(IgG)抗体、中和抗体活性(抑制浓度>50％[MN₅₀]的微中和测定)和人血管紧张素转化酶2(hACE2)受体结合抑制的测定。使用合格的IgG酶联免疫吸附测定(ELISA)检测对于SARS-CoV-2rS蛋白抗原具有特异性的血清IgG抗体水平。使用合格的野生型病毒MN测定测量对于SARS-CoV-2病毒具有特异性的中和抗体-。收集了原始和β变体SARS-CoV-2毒株二者的血清IgG和MN₅₀测定资料。符合目的的功能性hACE2抑制测定和抗rS(抗重组刺突)IgG活性测定二者均用于分析对SARS-CoV-2的原始毒株、B.1.351(β)、B.1.1.7(α)和B.1.617.2(δ)变异株的反应。

安全性结局包括在加强免疫之后7天内参与者报告的反应原性事件，以及截至加强免疫之后28天发生的非征集性不良事件。通过征集性局部和全身性不良事件分别记录了加强免疫反应原性。记录了从加强免疫疫苗接种到加强免疫之后28天的征集性不良事件。还收集了有关不良事件是否严重、是否与疫苗接种相关、是否与COVID-19相关、是否为潜在的免疫介导的医学病症(PIMMC)或是否导致中断或计划外就诊的资料。在加强免疫之前立即以及之后28天收集用于免疫原性分析的参与者样品。

统计资料：分析包括来自B组中的参与者的在初次疫苗接种系列期间和之后(第0天、第21天、第35天、第105天和第189天)获得的安全性和免疫原性数据，以用于与在接受加强剂量之后28天(第217天)从B2组收集的数据进行比较。结果还按参与者年龄组进行了分析：≥18至≤84岁、≥18至≤59岁和≥60至≤84岁。

安全性分析包括所有接受BV2373和皂苷佐剂的单次加强免疫注射(B2组)或安慰剂(B1组)的参与者。安全性分析表示为具有截至每次疫苗接种之后7天分析的征集性局部和全身性不良事件以及截至加强免疫之后28天非征集性不良事件的参与者的数量和百分比。

结果：共筛选1610名参与者。除了三名参与者之外全部被随机化为B组(n＝257)的参与者在他们的初次疫苗接种系列中接受了两个剂量的BV2373和皂苷佐剂，并被考虑进行相同剂量水平下单次加强剂量的研究(图63)。B组参与者在第189天重新随机化，其中210名同意的参与者按1:1分配以在B2组(n＝104)中接受BV2373和皂苷佐剂的单次加强免疫或在B1组(n＝106)中接受安慰剂。在B2组中，除一名参与者外，所有参与者都接受了作为加强免疫的活性疫苗。B1组中除六名参与者外所有参与者都接受了作为加强免疫的安慰剂；在其余六名参与者中，四名未接受任何加强免疫(其中一名因持续的不良事件而出于安全性的考虑被包括在B1组中)，并且两名参与者错误地接受了作为加强免疫的活性疫苗，并在B2组进行了安全性评估。A组中除一名参与者外，所有参与者都接受了全部三个剂量的安慰剂，且其余参与者接受了作为加强剂量的活性疫苗。

除了B1组(58％)中女性参与者的比例高于B2组(45％)之外，活性(B2组)与安慰剂(B1组)加强免疫组之间的人口统计学和基线特征总体上是平衡的(表8)。在A、B1和B2组中，中值年龄为约57岁，并且45％的参与者年龄≥60至≤84岁。大多数参与者是白人(87％)和非西班牙人或拉丁美洲人(95％)。基线SARS-CoV-2血清状态主要为阴性(98％)。

征集性局部和全身性反应原性事件的安全性报告显示在BV2373和皂苷佐剂的全部三个剂量下都是增加的趋势(图64A-图64B)。加强免疫后，B2组中的参与者报告任何局部反应(压痛、疼痛、肿胀和红斑)的发生率为82.5％(13.4％≥3级)，相比之下，在初次疫苗接种系列后为70.0％(5.2％≥3级)。4级局部反应很少见，且B2组中的一名参与者报告了两个事件(疼痛和压痛)，相比之下，在初次疫苗接种系列后没有参与者报告。加强免疫后，局部反应是短暂的，其中除红斑(2.5天)外，所有事件的中值持续时间为2.0天。在初次疫苗接种系列后，局部反应也是短暂，其中疼痛和压痛的中值持续时间为2.0天且红斑和肿胀的中值持续时间为1.0天。

全身性反应显示出相似的模式，其中任何事件(疲劳、头痛、肌肉疼痛、不适、关节疼痛、恶心/呕吐和发热)的发生率为76.5％(15.3％≥3级)，相比之下，在初次疫苗接种系列后为52.8％(5.6％≥3级)。4级全身性反应很少见，其中B2组中的一名参与者报告了三个事件(头痛、不适和肌肉疼痛)，相比之下，在初次疫苗接种系列后没有参与者报告。加强免疫后，全身性反应在本质上是短暂的，其中所有事件的中值持续时间为1.0天，但肌肉疼痛持续时间为2.0天。在初次疫苗接种系列后，所有全身性反应也是短暂的，其中所有事件的中值持续时间为1.0天。

当与较年轻的成人(≥18至≤59岁)相比时，在初次疫苗接种系列或加强剂量后，在较年长的成人(≥60至≤84岁)中的局部和全身性反应原性事件不太频繁且不太严重。在较年轻的群组中，分别在84.9％(18.9％≥3级)和84.9％(26.4％≥3级)的参与者中报告了加强免疫后的局部和全身性反应，相比之下，在较年长的群组中分别为79.5％(6.8％≥3级)和66.7％(2.2％≥3级)的参与者。

总计了在活性加强免疫的参与者(B2组)、安慰剂加强免疫的参与者(B1组)和在整个研究期间接受三个剂量的安慰剂的参与者(A组)中的征集性不良事件。截至加强免疫之后28天，与仅接受安慰剂的参与者(A组)相比，最初接受活性疫苗用于其初次疫苗接种系列的参与者(B2组和B1组)经历了征集性不良事件的更高发生率，分别有12.4％、12.7％、和11.0％的参与者报告了此类事件。对于非征集性重度不良事件也观察到类似的趋势(分别为5.7％、3.9％和2.4％)。报告的其他类型的AE包括医疗护理AE(需要医疗保健就诊的事件；MAAE)、潜在的免疫介导的医学病症(PIMMC)、与COVID 19相关的事件和严重不良事件(SAE)。

总体而言，三组中活性加强免疫的参与者发生MAAE的频率略高(B2、B1和A组分别为30.5％、26.1％和23.2％)，且少数参与者报告了相关事件(分别为1.9％、0％和1.2％)。在整个研究中，被认为是PIMMC的事件很少见，其中B2组和A组中的一名参与者各自报告了一个事件；这两个事件都被评估为与研究治疗无关。没有参与者报告与COVID-19相关的不良事件。

SAE在整个研究中也不常见，分别发生在B2、B1和A组中的5.7％、3.3％和1.6％的参与者中，且所有事件均被评估为与研究治疗无关。

对于B2和B1组参与者的SAE的评价未显示出与活性加强免疫的关系，因为在剂量1、剂量2和加强免疫之后SAE分别发生在B2组中的0％、4.8％和1.0％的参与者中以及B1组中的0％、2.0％和2.0％的参与者中。

在初次疫苗接种系列(第35天)至第189天后，观察到B组IgG和MN50几何平均滴度(GMT)的下降(分别地对于IgG，43,905个ELISA单位[EU]至6,064EU；且对于MN50，1,470至63)。加强免疫后二十八天(第217天)，IgG和MN50滴度与加强免疫前滴度和初次免疫系列产生的第35天滴度相比稳健地增加(图65，图66)。

对于原始SARS-CoV-2毒株，血清IgG GMT从初次疫苗接种系列(第35天)后的43,905EU增加至加强免疫(第217天)后的204,367EU，约4.7倍。与较年轻的成人(4.1倍)相比，在较年长的成人(5.1倍)中观察到加强免疫后的更高倍数增加。类似地，特定于原始SARS-CoV-2毒株的MN50测定GMT经历相同的各自时间点从1,470增加至6,023，约4.1倍；且在较年长的成人和较年轻的成人中分别为4.0倍和3.8倍的增加。

对于β变体，IgG GMT从加强免疫前第189天的4,317EU增加至第217天的175,190EU，反映了加强免疫后约40.6倍的增加。这些滴度比第35天对于原始毒株观察到的滴度高4倍(GMT 175,190EU相比于43,905EU)。β变体MN50测定数据显示，从加强免疫前(第189天)至加强免疫后(第217天)的滴度约50.1倍的相似倍数增加(GMT 13相比于661)，但滴度低于在第35天对于原始毒株所观察到的滴度(GMT 661相比于1,470)。(表9，表10)。

开发了两种测定以使用第35天(B组)和第217天(B2组)的参与者血清评估针对另外的SARS-CoV-2变体的免疫反应。使用功能性hACE2抑制测定来比较针对原始毒株(与SEQID NO:1相比，包含具有D614G突变的CoV S多肽的SARS-CoV-2病毒)以及SARS-CoV-2的δ、β和α变体的活性。以相应的顺序，观察到hACE2抑制滴度的6倍、6.6倍、10.8倍、8.1倍和19.9倍增加(表12A、表12B、图62A-图62B)。比较相同的SARS-CoV-2毒株之间的抗rS IgG活性的第二种测定发现，在加强免疫后观察到5.4倍(原始)、11.1倍(δ)、6.5倍(β)、9.7倍(α)更高的滴度(表11A、表11B、图61A-图61B)。

结果：在初次两个剂量系列后大约6个月，施用单次加强剂量的疫苗导致反应原性事件的增量式增加以及显著增强的免疫原性。

在第189天加强免疫之前，当与第35天初次疫苗接种系列后采集的样品相比时，免疫的参与者中抗SARS-CoV-2抗体滴度显著降低(B组IgG和MN₅₀ GMT分别从43,905EU降至6,064EU以及从1,470降至63)。中和抗体的存在强烈表明了保护免于症状性COVID-19的危害。

在本研究中，对于原始疫苗毒株以及最近的SARS-CoV-2变体(包括α、β和δ)，评估了对加强免疫的抗体反应。对于原始毒株，第217天的IgG滴度比加强免疫前第189天的滴度高约34倍，而加强免疫后的中和抗体滴度增加约96倍。加强免疫后的IgG和MN滴度均比第35天初次两剂量系列后观察到的滴度高>4倍，这是值得注意的，因为第35天的滴度对应于英国3期研究(89.7％)以及美国/墨西哥3期研究(90.4％)中的高水平的临床功效。按年龄组细分时，对于较年长的成人(≥60至≤84岁)观察到的倍数增加高于较年轻的成人(≥18至≤59岁)。这一发现表明，加强剂量可能在较年长的成人中有额外的益处，因为他们在初次两个剂量疫苗接种系列后的抗体反应低于在较年轻的成人中观察到的抗体反应。

对于β变体，在加强免疫后观察到IgG和MN抗体滴度的40至50倍增加，并且IgG滴度比初次疫苗接种系列后对于原始毒株所观察到的滴度高约4倍。与IgG的观察结果不同，加强免疫后β变体的MN₅₀ GMT低于初次疫苗接种系列后对于原始毒株的那些(GMT 661相比于1,470)，这与已知的对于该变体的降低的中和反应一致。

对于SARS-CoV-2的δ变体，当将加强免疫后第217天的滴度与第35天的滴度进行比较时，观察到功能性hACE2抑制滴度的6.6倍增加。在这些相同时间点比较的抗rS IgG活性发现高9.7倍(δ)的与加强免疫相关的滴度。

与先前的剂量相比，在第6个月加强剂量之后，局部和全身性反应原性的发生率均更高，这反映了在第三个剂量下观察到的增加的免疫原性。然而，3级或更高级别事件的发生率仍然相对较低，其中只有10％的参与者记录了疲劳(12.2％)。总共报告了五个4级(可能危及生命)征集性局部和全身性不良事件。所有这五个事件(疼痛、压痛、头痛、不适和肌肉疼痛)均由活性加强免疫组中的同一参与者报告，同时发生了与疫苗相关的药物超敏反应的不良事件。药物超敏反应事件的严重程度被评估为轻度。参与者没有为此事件寻求任何医疗护理，并且参与者的所有症状在6天的时间内消退。

表13显示了用于与SEQ ID NO:1相比具有D614G突变的SARS-CoV-2病毒或SARS-CoV-2δ变体的99％的中和的几何平均滴度。图67显示了实例11的包含BV2373和皂苷佐剂的免疫原性组合物针对抗含有D614G突变的SARS-CoV-2毒株和B.1.617.2(δ变体)的中和抗体99(neut99)值。

总体而言，在初次免疫系列之后约6个月施用的单次加强剂量的BV2373和皂苷佐剂诱导了体液抗体的显著增加，其比两项3期研究中与高水平功效相关的抗体滴度高>4倍，同时还显示出可接受的安全性特征。这些发现支持在加强免疫计划中使用所述疫苗。

表8：A、B1和B2组的人口统计学和基线特征

BMI＝体重指数；SARS-CoV-2＝严重急性呼吸综合征冠状病毒2；SD＝标准偏差A＝第0天、第21天和第189天安慰剂

B1＝第0天和第21天5μg BV2373+50μg皂苷佐剂以及第189天安慰剂

B2＝第0天、第21天和第189天5μg BV2373+50μg皂苷佐剂

1安全性分析组中的参与者根据所接受的治疗进行计数，以适应治疗错误。

表9：接受BV2373和皂苷佐剂的参与者按研究日针对原始和β变体SARS-CoV-2毒株的初次免疫和加强免疫疫苗接种后的血清IgG几何平均滴度

血清IgG GMT(EU[95％CI])

CI＝置信区间；ELISA＝酶联免疫吸附测定；EU＝ELISA单位；GMT＝几何平均滴度

表10：接受BV2373和皂苷佐剂的参与者按研究日针对原始和β变体SARS-CoV-2毒株的初次免疫和加强免疫疫苗接种后的中和抗体活性

MN₅₀ GMT(95％CI)

CI＝置信区间；GMT＝几何平均滴度；MN₅₀＝抑制浓度>50％下的微量中和测定

表11A：随时间而变的抗CoV S IgG

表11B：接受BV2373和皂苷佐剂的参与者按研究日针对原始和变体SARS-CoV-2毒株的初次免疫和加强免疫疫苗接种后的rS IgG几何平均滴度

表12A：随时间而变的50％hACE2抑制滴度

表12B：接受BV2373和皂苷佐剂的参与者按研究日针对原始和变体SARS-CoV-2毒株的初次免疫和加强免疫疫苗接种后的hACE2抑制几何平均滴度

表13：用于具有D614G突变的SARS-CoV-2病毒和B.1.617-2δ变体的中和的几何平均滴度

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序列表

<110> 诺瓦瓦克斯股份有限公司

<120> 冠状病毒疫苗制剂

<130> NOVV-092/01WO 305047-2992

<150> US 63/293,468

<151> 2021-12-23

<150> US 63/292,196

<151> 2021-12-21

<150> US 63/290,439

<151> 2021-12-16

<150> US 63/280,395

<151> 2021-11-17

<150> US 63/195,986

<151> 2021-06-02

<150> US 63/177,059

<151> 2021-04-20

<150> US 63/176,825

<151> 2021-04-19

<150> US 63/164,487

<151> 2021-03-22

<160> 168

<170> PatentIn version 3.5

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Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala

965 970 975

Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln

980 985 990

Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser

995 1000 1005

Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln

1010 1015 1020

Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser

1025 1030 1035

Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr

1040 1045 1050

Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile

1055 1060 1065

Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val

1070 1075 1080

Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu

1085 1090 1095

Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys

1100 1105 1110

Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu

1115 1120 1125

Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe

1130 1135 1140

Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly

1145 1150 1155

Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu

1160 1165 1170

Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln

1175 1180 1185

Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile

1190 1195 1200

Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr

1205 1210 1215

Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly

1220 1225 1230

Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser

1235 1240 1245

Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255 1260

<210> 3

<211> 1273

<212> PRT

<213> 人工序列

<220>

<223> 重组S蛋白

<400> 3

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln Ser Val Ala

675 680 685

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser

690 695 700

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

705 710 715 720

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

725 730 735

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

740 745 750

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

755 760 765

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

770 775 780

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe

785 790 795 800

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

805 810 815

Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly

820 825 830

Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp

835 840 845

Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu

850 855 860

Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly

865 870 875 880

Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile

885 890 895

Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr

900 905 910

Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn

915 920 925

Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala

930 935 940

Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn

945 950 955 960

Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val

965 970 975

Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln

980 985 990

Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val

995 1000 1005

Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn

1010 1015 1020

Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

1025 1030 1035

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro

1040 1045 1050

Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val

1055 1060 1065

Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

1070 1075 1080

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

1085 1090 1095

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln

1100 1105 1110

Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val

1115 1120 1125

Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro

1130 1135 1140

Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn

1145 1150 1155

His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn

1160 1165 1170

Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu

1175 1180 1185

Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu

1190 1195 1200

Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu

1205 1210 1215

Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met

1220 1225 1230

Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys

1235 1240 1245

Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro

1250 1255 1260

Val Leu Lys Gly Val Lys Leu His Tyr Thr

1265 1270

<210> 4

<211> 1260

<212> PRT

<213> 人工序列

<220>

<223> 重组S蛋白

<400> 4

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln

660 665 670

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala

675 680 685

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn

690 695 700

Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys

705 710 715 720

Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys

725 730 735

Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg

740 745 750

Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val

755 760 765

Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe

770 775 780

Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser

785 790 795 800

Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala

805 810 815

Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala

820 825 830

Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu

835 840 845

Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu

850 855 860

Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala

865 870 875 880

Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile

885 890 895

Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn

900 905 910

Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr

915 920 925

Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln

930 935 940

Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile

945 950 955 960

Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala

965 970 975

Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln

980 985 990

Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser

995 1000 1005

Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln

1010 1015 1020

Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser

1025 1030 1035

Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr

1040 1045 1050

Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile

1055 1060 1065

Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val

1070 1075 1080

Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu

1085 1090 1095

Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys

1100 1105 1110

Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu

1115 1120 1125

Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe

1130 1135 1140

Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly

1145 1150 1155

Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu

1160 1165 1170

Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln

1175 1180 1185

Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile

1190 1195 1200

Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr

1205 1210 1215

Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly

1220 1225 1230

Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser

1235 1240 1245

Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255 1260

<210> 5

<211> 13

<212> PRT

<213> 人工序列

<220>

<223> 信号肽

<400> 5

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser

1 5 10

<210> 6

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 6

Arg Arg Ala Arg

1

<210> 7

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 7

Gln Gln Ala Gln

1

<210> 8

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 8

Gln Arg Ala Arg

1

<210> 9

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 9

Arg Gln Ala Arg

1

<210> 10

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 10

Arg Arg Ala Gln

1

<210> 11

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 11

Gln Gln Ala Arg

1

<210> 12

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 12

Arg Gln Ala Gln

1

<210> 13

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 13

Gln Arg Ala Gln

1

<210> 14

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 14

Asn Asn Ala Asn

1

<210> 15

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 15

Asn Arg Ala Arg

1

<210> 16

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 16

Arg Asn Ala Arg

1

<210> 17

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 17

Arg Arg Ala Asn

1

<210> 18

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 18

Asn Asn Ala Arg

1

<210> 19

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 19

Arg Asn Ala Asn

1

<210> 20

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 20

Asn Arg Ala Asn

1

<210> 21

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 21

Ala Ala Ala Ala

1

<210> 22

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 22

Ala Arg Ala Arg

1

<210> 23

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 23

Arg Ala Ala Arg

1

<210> 24

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 24

Arg Arg Ala Ala

1

<210> 25

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 25

Ala Ala Ala Arg

1

<210> 26

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 26

Arg Ala Ala Ala

1

<210> 27

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 27

Ala Arg Ala Ala

1

<210> 28

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 28

Gly Gly Ala Gly

1

<210> 29

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 29

Gly Arg Ala Arg

1

<210> 30

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 30

Arg Gly Ala Arg

1

<210> 31

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 31

Arg Arg Ala Gly

1

<210> 32

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 32

Gly Gly Ala Arg

1

<210> 33

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 33

Arg Gly Ala Gly

1

<210> 34

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 34

Gly Arg Ala Gly

1

<210> 35

<211> 3819

<212> DNA

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 35

atgttcgtct tcctggtgct gctgcccctg gtgtccagcc agtgcgtgaa cctgaccact 60

aggactcagc tgcctcccgc ttacaccaac tcattcactc gcggtgtgta ctaccctgac 120

aaggtcttcc gttcttcagt gctgcactca actcaggacc tgttcctgcc cttcttctcc 180

aacgtcacct ggttccacgc catccacgtg tccggcacca acggcactaa gcgcttcgac 240

aacccagtgc tgcctttcaa cgacggtgtc tacttcgctt caaccgagaa gtccaacatc 300

atccgtggat ggatcttcgg caccactctg gacagcaaga ctcagtctct gctgatcgtc 360

aacaacgcca ccaacgtggt catcaaggtc tgcgaattcc agttctgcaa cgacccattc 420

ctgggcgtct actaccacaa gaacaacaag tcatggatgg agtccgaatt ccgcgtctac 480

tccagcgcta acaactgcac tttcgagtac gtgtcccagc ctttcctgat ggacctggaa 540

ggaaagcagg gtaacttcaa gaacctgagg gagttcgtgt tcaagaacat cgacggatac 600

ttcaagattt acagcaagca caccccaatc aacctggtgc gcgacctgcc tcagggtttc 660

tctgctctgg agccactggt ggacctgcct atcggcatca acatcacccg cttccagact 720

ctgctggctc tgcaccgttc ctacctgact ccaggcgact catcttctgg atggactgct 780

ggagctgctg cttactacgt gggctacctg cagcctcgca ccttcctgct gaagtacaac 840

gaaaacggaa ccatcactga cgccgtcgac tgcgctctgg accctctgtc agaaaccaag 900

tgcactctga agtccttcac cgtggagaag ggcatctacc agacttcaaa cttcagggtg 960

cagcccaccg aatccatcgt cagattccct aacatcacta acctgtgccc cttcggagag 1020

gtcttcaacg ccacccgctt cgcttccgtg tacgcctgga acaggaagag aatctcaaac 1080

tgcgtcgctg actactccgt gctgtacaac tcagcctcct tcagcacctt caagtgctac 1140

ggcgtgtcac caactaagct gaacgacctg tgcttcacca acgtctacgc cgactccttc 1200

gtgatcaggg gagacgaggt cagacagatc gctcctggcc agactggaaa gatcgccgac 1260

tacaactaca agctgcccga cgacttcacc ggttgcgtca tcgcttggaa cagcaacaac 1320

ctggactcta aagtgggtgg caactacaac tacctgtacc gcctgttccg taagtcaaac 1380

ctgaagccat tcgagaggga catcagcact gaaatctacc aggccggatc taccccttgc 1440

aacggtgtcg agggcttcaa ctgctacttc cccctgcagt cctacggttt ccagccaact 1500

aacggtgtgg gctaccagcc ttacagagtg gtcgtgctga gcttcgaact gctccacgct 1560

cctgctactg tgtgcggtcc aaagaagtct accaacctgg tcaagaacaa gtgcgtgaac 1620

ttcaacttca acggcctgac cggaactggt gtcctgaccg agagcaacaa gaagttcctg 1680

cccttccagc agttcggaag ggacatcgct gacaccactg acgctgtgcg cgaccctcag 1740

accctggaaa tcctggacat cactccatgc tcattcggag gtgtctccgt gatcacccct 1800

ggcaccaaca cttctaacca ggtcgctgtg ctgtaccagg acgtcaactg caccgaggtc 1860

cctgtggcca tccacgctga ccagctgacc cccacttggc gcgtgtactc caccggcagc 1920

aacgtgttcc agactcgtgc tggttgcctg atcggcgccg agcacgtgaa caacagctac 1980

gaatgcgaca tccccatcgg cgctggaatc tgcgcctctt accagaccca gactaacagc 2040

ccacgcaggg ctcgctctgt ggcctctcag tcaatcatcg cttacaccat gtcactgggc 2100

gctgaaaact ccgtggccta ctctaacaac tcaatcgcca tccccaccaa cttcactatc 2160

agcgtgacca ctgagatcct gccagtcagc atgaccaaga cttctgtgga ctgcactatg 2220

tacatctgcg gagacagcac cgaatgctct aacctgctgc tgcagtacgg ctctttctgc 2280

acccagctga accgtgctct gactggaatc gccgtggagc aggacaagaa cactcaggaa 2340

gtcttcgctc aggtgaagca aatctacaag accccaccta tcaaggactt cggcggattc 2400

aacttctccc agatcctgcc tgacccctcc aagccaagca agcgctcttt catcgaggac 2460

ctgctgttca acaaggtcac tctggccgac gctggattca tcaagcagta cggagactgc 2520

ctgggtgaca tcgccgctcg tgacctgatc tgcgctcaga agttcaacgg tctgaccgtg 2580

ctgcccccac tgctgactga cgaaatgatc gcccagtaca ctagcgccct gctggctgga 2640

accatcactt ctggttggac cttcggtgct ggcgccgctc tgcagatccc tttcgctatg 2700

cagatggcct accgtttcaa cggaatcggt gtcacccaga acgtgctgta cgagaaccag 2760

aagctgatcg ctaaccagtt caactcagcc atcggaaaga tccaggacag cctgagctct 2820

actgcctctg ctctgggcaa gctgcaggac gtcgtgaacc agaacgccca ggctctgaac 2880

accctggtca agcagctgtc atccaacttc ggtgctatca gctctgtgct gaacgacatc 2940

ctgtcccgcc tggacaaggt cgaggccgaa gtgcagatcg accgcctgat cactggccgt 3000

ctgcagtcac tgcagaccta cgtgactcag cagctgatca gggccgctga aatcagagcc 3060

tccgctaacc tggccgctac caagatgagc gagtgcgtcc tgggtcaatc taagcgtgtg 3120

gacttctgcg gcaagggata ccacctgatg tcattccctc agtctgctcc ccacggtgtg 3180

gtgttcctgc acgtcaccta cgtgccagcc caggagaaga acttcaccac tgcccctgct 3240

atctgccacg acggcaaggc tcacttcccc agggaaggtg tcttcgtgag caacggcacc 3300

cactggttcg tcactcagag aaacttctac gagccacaga tcatcaccac tgacaacact 3360

ttcgtgtctg gaaactgcga cgtggtcatc ggtatcgtca acaacaccgt gtacgacccc 3420

ctgcagccag agctggactc attcaaggag gaactggaca agtacttcaa gaaccacacc 3480

tcccctgacg tcgacctggg cgacatctca ggaatcaacg cttccgtcgt gaacatccag 3540

aaggagatcg accgcctgaa cgaagtggcc aagaacctga acgaaagcct gatcgacctg 3600

caggagctgg gcaagtacga acagtacatc aagtggcctt ggtacatctg gctgggtttc 3660

atcgctggcc tcatcgctat cgtgatggtg accatcatgc tgtgctgcat gacttcatgc 3720

tgctcctgcc tgaagggctg ctgcagctgc ggatcttgct gcaagttcga cgaggacgac 3780

tctgaacccg tcctgaaggg cgtgaagctg cactacacc 3819

<210> 36

<211> 1279

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 36

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg Ser Val Ala

675 680 685

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser

690 695 700

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

705 710 715 720

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

725 730 735

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

740 745 750

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

755 760 765

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

770 775 780

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe

785 790 795 800

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

805 810 815

Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly

820 825 830

Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp

835 840 845

Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu

850 855 860

Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly

865 870 875 880

Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile

885 890 895

Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr

900 905 910

Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn

915 920 925

Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala

930 935 940

Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn

945 950 955 960

Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val

965 970 975

Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln

980 985 990

Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val

995 1000 1005

Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn

1010 1015 1020

Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

1025 1030 1035

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro

1040 1045 1050

Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val

1055 1060 1065

Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

1070 1075 1080

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

1085 1090 1095

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln

1100 1105 1110

Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val

1115 1120 1125

Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro

1130 1135 1140

Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn

1145 1150 1155

His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn

1160 1165 1170

Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu

1175 1180 1185

Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu

1190 1195 1200

Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu

1205 1210 1215

Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met

1220 1225 1230

Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys

1235 1240 1245

Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro

1250 1255 1260

Val Leu Lys Gly Val Lys Leu His Tyr Thr His His His His His

1265 1270 1275

His

<210> 37

<211> 3837

<212> DNA

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 37

atgttcgtct tcctggtgct gctgcccctg gtgtccagcc agtgcgtgaa cctgaccact 60

aggactcagc tgcctcccgc ttacaccaac tcattcactc gcggtgtgta ctaccctgac 120

aaggtcttcc gttcttcagt gctgcactca actcaggacc tgttcctgcc cttcttctcc 180

aacgtcacct ggttccacgc catccacgtg tccggcacca acggcactaa gcgcttcgac 240

aacccagtgc tgcctttcaa cgacggtgtc tacttcgctt caaccgagaa gtccaacatc 300

atccgtggat ggatcttcgg caccactctg gacagcaaga ctcagtctct gctgatcgtc 360

aacaacgcca ccaacgtggt catcaaggtc tgcgaattcc agttctgcaa cgacccattc 420

ctgggcgtct actaccacaa gaacaacaag tcatggatgg agtccgaatt ccgcgtctac 480

tccagcgcta acaactgcac tttcgagtac gtgtcccagc ctttcctgat ggacctggaa 540

ggaaagcagg gtaacttcaa gaacctgagg gagttcgtgt tcaagaacat cgacggatac 600

ttcaagattt acagcaagca caccccaatc aacctggtgc gcgacctgcc tcagggtttc 660

tctgctctgg agccactggt ggacctgcct atcggcatca acatcacccg cttccagact 720

ctgctggctc tgcaccgttc ctacctgact ccaggcgact catcttctgg atggactgct 780

ggagctgctg cttactacgt gggctacctg cagcctcgca ccttcctgct gaagtacaac 840

gaaaacggaa ccatcactga cgccgtcgac tgcgctctgg accctctgtc agaaaccaag 900

tgcactctga agtccttcac cgtggagaag ggcatctacc agacttcaaa cttcagggtg 960

cagcccaccg aatccatcgt cagattccct aacatcacta acctgtgccc cttcggagag 1020

gtcttcaacg ccacccgctt cgcttccgtg tacgcctgga acaggaagag aatctcaaac 1080

tgcgtcgctg actactccgt gctgtacaac tcagcctcct tcagcacctt caagtgctac 1140

ggcgtgtcac caactaagct gaacgacctg tgcttcacca acgtctacgc cgactccttc 1200

gtgatcaggg gagacgaggt cagacagatc gctcctggcc agactggaaa gatcgccgac 1260

tacaactaca agctgcccga cgacttcacc ggttgcgtca tcgcttggaa cagcaacaac 1320

ctggactcta aagtgggtgg caactacaac tacctgtacc gcctgttccg taagtcaaac 1380

ctgaagccat tcgagaggga catcagcact gaaatctacc aggccggatc taccccttgc 1440

aacggtgtcg agggcttcaa ctgctacttc cccctgcagt cctacggttt ccagccaact 1500

aacggtgtgg gctaccagcc ttacagagtg gtcgtgctga gcttcgaact gctccacgct 1560

cctgctactg tgtgcggtcc aaagaagtct accaacctgg tcaagaacaa gtgcgtgaac 1620

ttcaacttca acggcctgac cggaactggt gtcctgaccg agagcaacaa gaagttcctg 1680

cccttccagc agttcggaag ggacatcgct gacaccactg acgctgtgcg cgaccctcag 1740

accctggaaa tcctggacat cactccatgc tcattcggag gtgtctccgt gatcacccct 1800

ggcaccaaca cttctaacca ggtcgctgtg ctgtaccagg acgtcaactg caccgaggtc 1860

cctgtggcca tccacgctga ccagctgacc cccacttggc gcgtgtactc caccggcagc 1920

aacgtgttcc agactcgtgc tggttgcctg atcggcgccg agcacgtgaa caacagctac 1980

gaatgcgaca tccccatcgg cgctggaatc tgcgcctctt accagaccca gactaacagc 2040

ccacgcaggg ctcgctctgt ggcctctcag tcaatcatcg cttacaccat gtcactgggc 2100

gctgaaaact ccgtggccta ctctaacaac tcaatcgcca tccccaccaa cttcactatc 2160

agcgtgacca ctgagatcct gccagtcagc atgaccaaga cttctgtgga ctgcactatg 2220

tacatctgcg gagacagcac cgaatgctct aacctgctgc tgcagtacgg ctctttctgc 2280

acccagctga accgtgctct gactggaatc gccgtggagc aggacaagaa cactcaggaa 2340

gtcttcgctc aggtgaagca aatctacaag accccaccta tcaaggactt cggcggattc 2400

aacttctccc agatcctgcc tgacccctcc aagccaagca agcgctcttt catcgaggac 2460

ctgctgttca acaaggtcac tctggccgac gctggattca tcaagcagta cggagactgc 2520

ctgggtgaca tcgccgctcg tgacctgatc tgcgctcaga agttcaacgg tctgaccgtg 2580

ctgcccccac tgctgactga cgaaatgatc gcccagtaca ctagcgccct gctggctgga 2640

accatcactt ctggttggac cttcggtgct ggcgccgctc tgcagatccc tttcgctatg 2700

cagatggcct accgtttcaa cggaatcggt gtcacccaga acgtgctgta cgagaaccag 2760

aagctgatcg ctaaccagtt caactcagcc atcggaaaga tccaggacag cctgagctct 2820

actgcctctg ctctgggcaa gctgcaggac gtcgtgaacc agaacgccca ggctctgaac 2880

accctggtca agcagctgtc atccaacttc ggtgctatca gctctgtgct gaacgacatc 2940

ctgtcccgcc tggacaaggt cgaggccgaa gtgcagatcg accgcctgat cactggccgt 3000

ctgcagtcac tgcagaccta cgtgactcag cagctgatca gggccgctga aatcagagcc 3060

tccgctaacc tggccgctac caagatgagc gagtgcgtcc tgggtcaatc taagcgtgtg 3120

gacttctgcg gcaagggata ccacctgatg tcattccctc agtctgctcc ccacggtgtg 3180

gtgttcctgc acgtcaccta cgtgccagcc caggagaaga acttcaccac tgcccctgct 3240

atctgccacg acggcaaggc tcacttcccc agggaaggtg tcttcgtgag caacggcacc 3300

cactggttcg tcactcagag aaacttctac gagccacaga tcatcaccac tgacaacact 3360

ttcgtgtctg gaaactgcga cgtggtcatc ggtatcgtca acaacaccgt gtacgacccc 3420

ctgcagccag agctggactc attcaaggag gaactggaca agtacttcaa gaaccacacc 3480

tcccctgacg tcgacctggg cgacatctca ggaatcaacg cttccgtcgt gaacatccag 3540

aaggagatcg accgcctgaa cgaagtggcc aagaacctga acgaaagcct gatcgacctg 3600

caggagctgg gcaagtacga acagtacatc aagtggcctt ggtacatctg gctgggtttc 3660

atcgctggcc tcatcgctat cgtgatggtg accatcatgc tgtgctgcat gacttcatgc 3720

tgctcctgcc tgaagggctg ctgcagctgc ggatcttgct gcaagttcga cgaggacgac 3780

tctgaacccg tcctgaaggg cgtgaagctg cactacaccc accaccacca ccaccac 3837

<210> 38

<211> 1266

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 38

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg

660 665 670

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala

675 680 685

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn

690 695 700

Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys

705 710 715 720

Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys

725 730 735

Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg

740 745 750

Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val

755 760 765

Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe

770 775 780

Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser

785 790 795 800

Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala

805 810 815

Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala

820 825 830

Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu

835 840 845

Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu

850 855 860

Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala

865 870 875 880

Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile

885 890 895

Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn

900 905 910

Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr

915 920 925

Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln

930 935 940

Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile

945 950 955 960

Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala

965 970 975

Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln

980 985 990

Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser

995 1000 1005

Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln

1010 1015 1020

Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser

1025 1030 1035

Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr

1040 1045 1050

Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile

1055 1060 1065

Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val

1070 1075 1080

Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu

1085 1090 1095

Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys

1100 1105 1110

Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu

1115 1120 1125

Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe

1130 1135 1140

Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly

1145 1150 1155

Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu

1160 1165 1170

Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln

1175 1180 1185

Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile

1190 1195 1200

Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr

1205 1210 1215

Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly

1220 1225 1230

Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser

1235 1240 1245

Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr His His His

1250 1255 1260

His His His

1265

<210> 39

<211> 60

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 39

Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met

1 5 10 15

Val Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys

20 25 30

Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser

35 40 45

Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

50 55 60

<210> 40

<211> 1213

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 40

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg Ser Val Ala

675 680 685

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser

690 695 700

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

705 710 715 720

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

725 730 735

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

740 745 750

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

755 760 765

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

770 775 780

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe

785 790 795 800

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

805 810 815

Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly

820 825 830

Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp

835 840 845

Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu

850 855 860

Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly

865 870 875 880

Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile

885 890 895

Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr

900 905 910

Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn

915 920 925

Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala

930 935 940

Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn

945 950 955 960

Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val

965 970 975

Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln

980 985 990

Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val

995 1000 1005

Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn

1010 1015 1020

Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

1025 1030 1035

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro

1040 1045 1050

Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val

1055 1060 1065

Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

1070 1075 1080

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

1085 1090 1095

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln

1100 1105 1110

Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val

1115 1120 1125

Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro

1130 1135 1140

Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn

1145 1150 1155

His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn

1160 1165 1170

Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu

1175 1180 1185

Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu

1190 1195 1200

Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro

1205 1210

<210> 41

<211> 1200

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 41

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg

660 665 670

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala

675 680 685

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn

690 695 700

Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys

705 710 715 720

Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys

725 730 735

Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg

740 745 750

Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val

755 760 765

Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe

770 775 780

Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser

785 790 795 800

Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala

805 810 815

Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala

820 825 830

Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu

835 840 845

Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu

850 855 860

Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala

865 870 875 880

Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile

885 890 895

Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn

900 905 910

Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr

915 920 925

Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln

930 935 940

Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile

945 950 955 960

Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala

965 970 975

Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln

980 985 990

Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser

995 1000 1005

Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln

1010 1015 1020

Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser

1025 1030 1035

Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr

1040 1045 1050

Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile

1055 1060 1065

Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val

1070 1075 1080

Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu

1085 1090 1095

Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys

1100 1105 1110

Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu

1115 1120 1125

Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe

1130 1135 1140

Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly

1145 1150 1155

Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu

1160 1165 1170

Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln

1175 1180 1185

Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro

1190 1195 1200

<210> 42

<211> 1219

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 42

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg Ser Val Ala

675 680 685

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser

690 695 700

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

705 710 715 720

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

725 730 735

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

740 745 750

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

755 760 765

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

770 775 780

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe

785 790 795 800

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

805 810 815

Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly

820 825 830

Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp

835 840 845

Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu

850 855 860

Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly

865 870 875 880

Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile

885 890 895

Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr

900 905 910

Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn

915 920 925

Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala

930 935 940

Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn

945 950 955 960

Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val

965 970 975

Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln

980 985 990

Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val

995 1000 1005

Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn

1010 1015 1020

Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

1025 1030 1035

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro

1040 1045 1050

Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val

1055 1060 1065

Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

1070 1075 1080

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

1085 1090 1095

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln

1100 1105 1110

Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val

1115 1120 1125

Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro

1130 1135 1140

Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn

1145 1150 1155

His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn

1160 1165 1170

Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu

1175 1180 1185

Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu

1190 1195 1200

Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro His His His His His

1205 1210 1215

His

<210> 43

<211> 3657

<212> DNA

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 43

atgttcgtct tcctggtgct gctgcccctg gtgtccagcc agtgcgtgaa cctgaccact 60

aggactcagc tgcctcccgc ttacaccaac tcattcactc gcggtgtgta ctaccctgac 120

aaggtcttcc gttcttcagt gctgcactca actcaggacc tgttcctgcc cttcttctcc 180

aacgtcacct ggttccacgc catccacgtg tccggcacca acggcactaa gcgcttcgac 240

aacccagtgc tgcctttcaa cgacggtgtc tacttcgctt caaccgagaa gtccaacatc 300

atccgtggat ggatcttcgg caccactctg gacagcaaga ctcagtctct gctgatcgtc 360

aacaacgcca ccaacgtggt catcaaggtc tgcgaattcc agttctgcaa cgacccattc 420

ctgggcgtct actaccacaa gaacaacaag tcatggatgg agtccgaatt ccgcgtctac 480

tccagcgcta acaactgcac tttcgagtac gtgtcccagc ctttcctgat ggacctggaa 540

ggaaagcagg gtaacttcaa gaacctgagg gagttcgtgt tcaagaacat cgacggatac 600

ttcaagattt acagcaagca caccccaatc aacctggtgc gcgacctgcc tcagggtttc 660

tctgctctgg agccactggt ggacctgcct atcggcatca acatcacccg cttccagact 720

ctgctggctc tgcaccgttc ctacctgact ccaggcgact catcttctgg atggactgct 780

ggagctgctg cttactacgt gggctacctg cagcctcgca ccttcctgct gaagtacaac 840

gaaaacggaa ccatcactga cgccgtcgac tgcgctctgg accctctgtc agaaaccaag 900

tgcactctga agtccttcac cgtggagaag ggcatctacc agacttcaaa cttcagggtg 960

cagcccaccg aatccatcgt cagattccct aacatcacta acctgtgccc cttcggagag 1020

gtcttcaacg ccacccgctt cgcttccgtg tacgcctgga acaggaagag aatctcaaac 1080

tgcgtcgctg actactccgt gctgtacaac tcagcctcct tcagcacctt caagtgctac 1140

ggcgtgtcac caactaagct gaacgacctg tgcttcacca acgtctacgc cgactccttc 1200

gtgatcaggg gagacgaggt cagacagatc gctcctggcc agactggaaa gatcgccgac 1260

tacaactaca agctgcccga cgacttcacc ggttgcgtca tcgcttggaa cagcaacaac 1320

ctggactcta aagtgggtgg caactacaac tacctgtacc gcctgttccg taagtcaaac 1380

ctgaagccat tcgagaggga catcagcact gaaatctacc aggccggatc taccccttgc 1440

aacggtgtcg agggcttcaa ctgctacttc cccctgcagt cctacggttt ccagccaact 1500

aacggtgtgg gctaccagcc ttacagagtg gtcgtgctga gcttcgaact gctccacgct 1560

cctgctactg tgtgcggtcc aaagaagtct accaacctgg tcaagaacaa gtgcgtgaac 1620

ttcaacttca acggcctgac cggaactggt gtcctgaccg agagcaacaa gaagttcctg 1680

cccttccagc agttcggaag ggacatcgct gacaccactg acgctgtgcg cgaccctcag 1740

accctggaaa tcctggacat cactccatgc tcattcggag gtgtctccgt gatcacccct 1800

ggcaccaaca cttctaacca ggtcgctgtg ctgtaccagg acgtcaactg caccgaggtc 1860

cctgtggcca tccacgctga ccagctgacc cccacttggc gcgtgtactc caccggcagc 1920

aacgtgttcc agactcgtgc tggttgcctg atcggcgccg agcacgtgaa caacagctac 1980

gaatgcgaca tccccatcgg cgctggaatc tgcgcctctt accagaccca gactaacagc 2040

ccacgcaggg ctcgctctgt ggcctctcag tcaatcatcg cttacaccat gtcactgggc 2100

gctgaaaact ccgtggccta ctctaacaac tcaatcgcca tccccaccaa cttcactatc 2160

agcgtgacca ctgagatcct gccagtcagc atgaccaaga cttctgtgga ctgcactatg 2220

tacatctgcg gagacagcac cgaatgctct aacctgctgc tgcagtacgg ctctttctgc 2280

acccagctga accgtgctct gactggaatc gccgtggagc aggacaagaa cactcaggaa 2340

gtcttcgctc aggtgaagca aatctacaag accccaccta tcaaggactt cggcggattc 2400

aacttctccc agatcctgcc tgacccctcc aagccaagca agcgctcttt catcgaggac 2460

ctgctgttca acaaggtcac tctggccgac gctggattca tcaagcagta cggagactgc 2520

ctgggtgaca tcgccgctcg tgacctgatc tgcgctcaga agttcaacgg tctgaccgtg 2580

ctgcccccac tgctgactga cgaaatgatc gcccagtaca ctagcgccct gctggctgga 2640

accatcactt ctggttggac cttcggtgct ggcgccgctc tgcagatccc tttcgctatg 2700

cagatggcct accgtttcaa cggaatcggt gtcacccaga acgtgctgta cgagaaccag 2760

aagctgatcg ctaaccagtt caactcagcc atcggaaaga tccaggacag cctgagctct 2820

actgcctctg ctctgggcaa gctgcaggac gtcgtgaacc agaacgccca ggctctgaac 2880

accctggtca agcagctgtc atccaacttc ggtgctatca gctctgtgct gaacgacatc 2940

ctgtcccgcc tggacaaggt cgaggccgaa gtgcagatcg accgcctgat cactggccgt 3000

ctgcagtcac tgcagaccta cgtgactcag cagctgatca gggccgctga aatcagagcc 3060

tccgctaacc tggccgctac caagatgagc gagtgcgtcc tgggtcaatc taagcgtgtg 3120

gacttctgcg gcaagggata ccacctgatg tcattccctc agtctgctcc ccacggtgtg 3180

gtgttcctgc acgtcaccta cgtgccagcc caggagaaga acttcaccac tgcccctgct 3240

atctgccacg acggcaaggc tcacttcccc agggaaggtg tcttcgtgag caacggcacc 3300

cactggttcg tcactcagag aaacttctac gagccacaga tcatcaccac tgacaacact 3360

ttcgtgtctg gaaactgcga cgtggtcatc ggtatcgtca acaacaccgt gtacgacccc 3420

ctgcagccag agctggactc attcaaggag gaactggaca agtacttcaa gaaccacacc 3480

tcccctgacg tcgacctggg cgacatctca ggaatcaacg cttccgtcgt gaacatccag 3540

aaggagatcg accgcctgaa cgaagtggcc aagaacctga acgaaagcct gatcgacctg 3600

caggagctgg gcaagtacga acagtacatc aagtggcctc accaccacca ccaccac 3657

<210> 44

<211> 1219

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 44

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg Ser Val Ala

675 680 685

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser

690 695 700

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

705 710 715 720

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

725 730 735

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

740 745 750

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

755 760 765

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

770 775 780

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe

785 790 795 800

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

805 810 815

Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly

820 825 830

Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp

835 840 845

Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu

850 855 860

Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly

865 870 875 880

Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile

885 890 895

Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr

900 905 910

Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn

915 920 925

Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala

930 935 940

Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn

945 950 955 960

Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val

965 970 975

Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln

980 985 990

Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val

995 1000 1005

Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn

1010 1015 1020

Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

1025 1030 1035

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro

1040 1045 1050

Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val

1055 1060 1065

Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

1070 1075 1080

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

1085 1090 1095

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln

1100 1105 1110

Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val

1115 1120 1125

Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro

1130 1135 1140

Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn

1145 1150 1155

His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn

1160 1165 1170

Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu

1175 1180 1185

Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu

1190 1195 1200

Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro His His His His His

1205 1210 1215

His

<210> 45

<211> 318

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 45

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn

305 310 315

<210> 46

<211> 955

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 46

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Ile Thr Asn

1 5 10 15

Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val

20 25 30

Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser

35 40 45

Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val

50 55 60

Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp

65 70 75 80

Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln

85 90 95

Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr

100 105 110

Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly

115 120 125

Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys

130 135 140

Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr

145 150 155 160

Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser

165 170 175

Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val

180 185 190

Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly

195 200 205

Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn

210 215 220

Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys

225 230 235 240

Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp

245 250 255

Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys

260 265 270

Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn

275 280 285

Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val

290 295 300

Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr

305 310 315 320

Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu

325 330 335

His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile

340 345 350

Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg Ser

355 360 365

Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu

370 375 380

Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe

385 390 395 400

Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr

405 410 415

Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser

420 425 430

Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala

435 440 445

Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe

450 455 460

Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly

465 470 475 480

Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys

485 490 495

Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp

500 505 510

Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala

515 520 525

Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro

530 535 540

Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu

545 550 555 560

Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu

565 570 575

Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly

580 585 590

Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln

595 600 605

Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala

610 615 620

Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala

625 630 635 640

Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser

645 650 655

Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu

660 665 670

Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr

675 680 685

Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala

690 695 700

Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

705 710 715 720

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln

725 730 735

Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala

740 745 750

Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys

755 760 765

Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His Trp

770 775 780

Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp

785 790 795 800

Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn

805 810 815

Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu

820 825 830

Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu

835 840 845

Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu

850 855 860

Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile

865 870 875 880

Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp

885 890 895

Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val

900 905 910

Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly

915 920 925

Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu

930 935 940

Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

945 950 955

<210> 47

<211> 2868

<212> DNA

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 47

atgttcgtct tcctggtgct gctgcccctg gtgtccagca tcactaacct gtgccccttc 60

ggagaggtct tcaacgccac ccgcttcgct tccgtgtacg cctggaacag gaagagaatc 120

tcaaactgcg tcgctgacta ctccgtgctg tacaactcag cctccttcag caccttcaag 180

tgctacggcg tgtcaccaac taagctgaac gacctgtgct tcaccaacgt ctacgccgac 240

tccttcgtga tcaggggaga cgaggtcaga cagatcgctc ctggccagac tggaaagatc 300

gccgactaca actacaagct gcccgacgac ttcaccggtt gcgtcatcgc ttggaacagc 360

aacaacctgg actctaaagt gggtggcaac tacaactacc tgtaccgcct gttccgtaag 420

tcaaacctga agccattcga gagggacatc agcactgaaa tctaccaggc cggatctacc 480

ccttgcaacg gtgtcgaggg cttcaactgc tacttccccc tgcagtccta cggtttccag 540

ccaactaacg gtgtgggcta ccagccttac agagtggtcg tgctgagctt cgaactgctc 600

cacgctcctg ctactgtgtg cggtccaaag aagtctacca acctggtcaa gaacaagtgc 660

gtgaacttca acttcaacgg cctgaccgga actggtgtcc tgaccgagag caacaagaag 720

ttcctgccct tccagcagtt cggaagggac atcgctgaca ccactgacgc tgtgcgcgac 780

cctcagaccc tggaaatcct ggacatcact ccatgctcat tcggaggtgt ctccgtgatc 840

acccctggca ccaacacttc taaccaggtc gctgtgctgt accaggacgt caactgcacc 900

gaggtccctg tggccatcca cgctgaccag ctgaccccca cttggcgcgt gtactccacc 960

ggcagcaacg tgttccagac tcgtgctggt tgcctgatcg gcgccgagca cgtgaacaac 1020

agctacgaat gcgacatccc catcggcgct ggaatctgcg cctcttacca gacccagact 1080

aacagcccac gcagggctcg ctctgtggcc tctcagtcaa tcatcgctta caccatgtca 1140

ctgggcgctg aaaactccgt ggcctactct aacaactcaa tcgccatccc caccaacttc 1200

actatcagcg tgaccactga gatcctgcca gtcagcatga ccaagacttc tgtggactgc 1260

actatgtaca tctgcggaga cagcaccgaa tgctctaacc tgctgctgca gtacggctct 1320

ttctgcaccc agctgaaccg tgctctgact ggaatcgccg tggagcagga caagaacact 1380

caggaagtct tcgctcaggt gaagcaaatc tacaagaccc cacctatcaa ggacttcggc 1440

ggattcaact tctcccagat cctgcctgac ccctccaagc caagcaagcg ctctttcatc 1500

gaggacctgc tgttcaacaa ggtcactctg gccgacgctg gattcatcaa gcagtacgga 1560

gactgcctgg gtgacatcgc cgctcgtgac ctgatctgcg ctcagaagtt caacggtctg 1620

accgtgctgc ccccactgct gactgacgaa atgatcgccc agtacactag cgccctgctg 1680

gctggaacca tcacttctgg ttggaccttc ggtgctggcg ccgctctgca gatccctttc 1740

gctatgcaga tggcctaccg tttcaacgga atcggtgtca cccagaacgt gctgtacgag 1800

aaccagaagc tgatcgctaa ccagttcaac tcagccatcg gaaagatcca ggacagcctg 1860

agctctactg cctctgctct gggcaagctg caggacgtcg tgaaccagaa cgcccaggct 1920

ctgaacaccc tggtcaagca gctgtcatcc aacttcggtg ctatcagctc tgtgctgaac 1980

gacatcctgt cccgcctgga caaggtcgag gccgaagtgc agatcgaccg cctgatcact 2040

ggccgtctgc agtcactgca gacctacgtg actcagcagc tgatcagggc cgctgaaatc 2100

agagcctccg ctaacctggc cgctaccaag atgagcgagt gcgtcctggg tcaatctaag 2160

cgtgtggact tctgcggcaa gggataccac ctgatgtcat tccctcagtc tgctccccac 2220

ggtgtggtgt tcctgcacgt cacctacgtg ccagcccagg agaagaactt caccactgcc 2280

cctgctatct gccacgacgg caaggctcac ttccccaggg aaggtgtctt cgtgagcaac 2340

ggcacccact ggttcgtcac tcagagaaac ttctacgagc cacagatcat caccactgac 2400

aacactttcg tgtctggaaa ctgcgacgtg gtcatcggta tcgtcaacaa caccgtgtac 2460

gaccccctgc agccagagct ggactcattc aaggaggaac tggacaagta cttcaagaac 2520

cacacctccc ctgacgtcga cctgggcgac atctcaggaa tcaacgcttc cgtcgtgaac 2580

atccagaagg agatcgaccg cctgaacgaa gtggccaaga acctgaacga aagcctgatc 2640

gacctgcagg agctgggcaa gtacgaacag tacatcaagt ggccttggta catctggctg 2700

ggtttcatcg ctggcctcat cgctatcgtg atggtgacca tcatgctgtg ctgcatgact 2760

tcatgctgct cctgcctgaa gggctgctgc agctgcggat cttgctgcaa gttcgacgag 2820

gacgactctg aacccgtcct gaagggcgtg aagctgcact acacctaa 2868

<210> 48

<211> 942

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 48

Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe

1 5 10 15

Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala

20 25 30

Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys

35 40 45

Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val

50 55 60

Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala

65 70 75 80

Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp

85 90 95

Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser

100 105 110

Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser

115 120 125

Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala

130 135 140

Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro

145 150 155 160

Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro

165 170 175

Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr

180 185 190

Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val

195 200 205

Asn Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser

210 215 220

Asn Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp

225 230 235 240

Thr Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile

245 250 255

Thr Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn

260 265 270

Thr Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu

275 280 285

Val Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val

290 295 300

Tyr Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile

305 310 315 320

Gly Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly

325 330 335

Ala Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg

340 345 350

Ala Arg Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu

355 360 365

Gly Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro

370 375 380

Thr Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met

385 390 395 400

Thr Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr

405 410 415

Glu Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu

420 425 430

Asn Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln

435 440 445

Glu Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys

450 455 460

Asp Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys

465 470 475 480

Pro Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr

485 490 495

Leu Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp

500 505 510

Ile Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr

515 520 525

Val Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser

530 535 540

Ala Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly

545 550 555 560

Ala Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn

565 570 575

Gly Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile

580 585 590

Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser

595 600 605

Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn

610 615 620

Ala Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly

625 630 635 640

Ala Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val

645 650 655

Glu Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser

660 665 670

Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg

675 680 685

Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly

690 695 700

Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser

705 710 715 720

Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr

725 730 735

Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

740 745 750

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly

755 760 765

Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile

770 775 780

Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly

785 790 795 800

Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser

805 810 815

Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp

820 825 830

Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile

835 840 845

Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu

850 855 860

Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys

865 870 875 880

Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile

885 890 895

Val Met Val Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys

900 905 910

Leu Lys Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp

915 920 925

Asp Ser Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

930 935 940

<210> 49

<211> 960

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 49

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Ile Thr Asn

1 5 10 15

Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val

20 25 30

Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser

35 40 45

Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val

50 55 60

Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp

65 70 75 80

Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln

85 90 95

Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr

100 105 110

Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly

115 120 125

Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys

130 135 140

Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr

145 150 155 160

Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser

165 170 175

Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val

180 185 190

Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly

195 200 205

Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn

210 215 220

Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys

225 230 235 240

Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp

245 250 255

Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys

260 265 270

Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn

275 280 285

Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val

290 295 300

Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr

305 310 315 320

Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu

325 330 335

His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile

340 345 350

Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg Ser

355 360 365

Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu

370 375 380

Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe

385 390 395 400

Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr

405 410 415

Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser

420 425 430

Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala

435 440 445

Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe

450 455 460

Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly

465 470 475 480

Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys

485 490 495

Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp

500 505 510

Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala

515 520 525

Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro

530 535 540

Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu

545 550 555 560

Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu

565 570 575

Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly

580 585 590

Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln

595 600 605

Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala

610 615 620

Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala

625 630 635 640

Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser

645 650 655

Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu

660 665 670

Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr

675 680 685

Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala

690 695 700

Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

705 710 715 720

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln

725 730 735

Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala

740 745 750

Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys

755 760 765

Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His Trp

770 775 780

Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp

785 790 795 800

Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn

805 810 815

Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu

820 825 830

Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu

835 840 845

Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu

850 855 860

Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile

865 870 875 880

Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp

885 890 895

Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val

900 905 910

Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly

915 920 925

Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu

930 935 940

Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr His His His His His

945 950 955 960

<210> 50

<211> 2883

<212> DNA

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 50

atgttcgtct tcctggtgct gctgcccctg gtgtccagca tcactaacct gtgccccttc 60

ggagaggtct tcaacgccac ccgcttcgct tccgtgtacg cctggaacag gaagagaatc 120

tcaaactgcg tcgctgacta ctccgtgctg tacaactcag cctccttcag caccttcaag 180

tgctacggcg tgtcaccaac taagctgaac gacctgtgct tcaccaacgt ctacgccgac 240

tccttcgtga tcaggggaga cgaggtcaga cagatcgctc ctggccagac tggaaagatc 300

gccgactaca actacaagct gcccgacgac ttcaccggtt gcgtcatcgc ttggaacagc 360

aacaacctgg actctaaagt gggtggcaac tacaactacc tgtaccgcct gttccgtaag 420

tcaaacctga agccattcga gagggacatc agcactgaaa tctaccaggc cggatctacc 480

ccttgcaacg gtgtcgaggg cttcaactgc tacttccccc tgcagtccta cggtttccag 540

ccaactaacg gtgtgggcta ccagccttac agagtggtcg tgctgagctt cgaactgctc 600

cacgctcctg ctactgtgtg cggtccaaag aagtctacca acctggtcaa gaacaagtgc 660

gtgaacttca acttcaacgg cctgaccgga actggtgtcc tgaccgagag caacaagaag 720

ttcctgccct tccagcagtt cggaagggac atcgctgaca ccactgacgc tgtgcgcgac 780

cctcagaccc tggaaatcct ggacatcact ccatgctcat tcggaggtgt ctccgtgatc 840

acccctggca ccaacacttc taaccaggtc gctgtgctgt accaggacgt caactgcacc 900

gaggtccctg tggccatcca cgctgaccag ctgaccccca cttggcgcgt gtactccacc 960

ggcagcaacg tgttccagac tcgtgctggt tgcctgatcg gcgccgagca cgtgaacaac 1020

agctacgaat gcgacatccc catcggcgct ggaatctgcg cctcttacca gacccagact 1080

aacagcccac gcagggctcg ctctgtggcc tctcagtcaa tcatcgctta caccatgtca 1140

ctgggcgctg aaaactccgt ggcctactct aacaactcaa tcgccatccc caccaacttc 1200

actatcagcg tgaccactga gatcctgcca gtcagcatga ccaagacttc tgtggactgc 1260

actatgtaca tctgcggaga cagcaccgaa tgctctaacc tgctgctgca gtacggctct 1320

ttctgcaccc agctgaaccg tgctctgact ggaatcgccg tggagcagga caagaacact 1380

caggaagtct tcgctcaggt gaagcaaatc tacaagaccc cacctatcaa ggacttcggc 1440

ggattcaact tctcccagat cctgcctgac ccctccaagc caagcaagcg ctctttcatc 1500

gaggacctgc tgttcaacaa ggtcactctg gccgacgctg gattcatcaa gcagtacgga 1560

gactgcctgg gtgacatcgc cgctcgtgac ctgatctgcg ctcagaagtt caacggtctg 1620

accgtgctgc ccccactgct gactgacgaa atgatcgccc agtacactag cgccctgctg 1680

gctggaacca tcacttctgg ttggaccttc ggtgctggcg ccgctctgca gatccctttc 1740

gctatgcaga tggcctaccg tttcaacgga atcggtgtca cccagaacgt gctgtacgag 1800

aaccagaagc tgatcgctaa ccagttcaac tcagccatcg gaaagatcca ggacagcctg 1860

agctctactg cctctgctct gggcaagctg caggacgtcg tgaaccagaa cgcccaggct 1920

ctgaacaccc tggtcaagca gctgtcatcc aacttcggtg ctatcagctc tgtgctgaac 1980

gacatcctgt cccgcctgga caaggtcgag gccgaagtgc agatcgaccg cctgatcact 2040

ggccgtctgc agtcactgca gacctacgtg actcagcagc tgatcagggc cgctgaaatc 2100

agagcctccg ctaacctggc cgctaccaag atgagcgagt gcgtcctggg tcaatctaag 2160

cgtgtggact tctgcggcaa gggataccac ctgatgtcat tccctcagtc tgctccccac 2220

ggtgtggtgt tcctgcacgt cacctacgtg ccagcccagg agaagaactt caccactgcc 2280

cctgctatct gccacgacgg caaggctcac ttccccaggg aaggtgtctt cgtgagcaac 2340

ggcacccact ggttcgtcac tcagagaaac ttctacgagc cacagatcat caccactgac 2400

aacactttcg tgtctggaaa ctgcgacgtg gtcatcggta tcgtcaacaa caccgtgtac 2460

gaccccctgc agccagagct ggactcattc aaggaggaac tggacaagta cttcaagaac 2520

cacacctccc ctgacgtcga cctgggcgac atctcaggaa tcaacgcttc cgtcgtgaac 2580

atccagaagg agatcgaccg cctgaacgaa gtggccaaga acctgaacga aagcctgatc 2640

gacctgcagg agctgggcaa gtacgaacag tacatcaagt ggccttggta catctggctg 2700

ggtttcatcg ctggcctcat cgctatcgtg atggtgacca tcatgctgtg ctgcatgact 2760

tcatgctgct cctgcctgaa gggctgctgc agctgcggat cttgctgcaa gttcgacgag 2820

gacgactctg aacccgtcct gaagggcgtg aagctgcact acacccacca ccaccaccac 2880

cac 2883

<210> 51

<211> 947

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 51

Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe

1 5 10 15

Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala

20 25 30

Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys

35 40 45

Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val

50 55 60

Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala

65 70 75 80

Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp

85 90 95

Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser

100 105 110

Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser

115 120 125

Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala

130 135 140

Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro

145 150 155 160

Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro

165 170 175

Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr

180 185 190

Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val

195 200 205

Asn Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser

210 215 220

Asn Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp

225 230 235 240

Thr Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile

245 250 255

Thr Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn

260 265 270

Thr Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu

275 280 285

Val Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val

290 295 300

Tyr Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile

305 310 315 320

Gly Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly

325 330 335

Ala Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg

340 345 350

Ala Arg Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu

355 360 365

Gly Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro

370 375 380

Thr Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met

385 390 395 400

Thr Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr

405 410 415

Glu Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu

420 425 430

Asn Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln

435 440 445

Glu Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys

450 455 460

Asp Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys

465 470 475 480

Pro Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr

485 490 495

Leu Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp

500 505 510

Ile Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr

515 520 525

Val Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser

530 535 540

Ala Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly

545 550 555 560

Ala Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn

565 570 575

Gly Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile

580 585 590

Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser

595 600 605

Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn

610 615 620

Ala Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly

625 630 635 640

Ala Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val

645 650 655

Glu Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser

660 665 670

Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg

675 680 685

Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly

690 695 700

Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser

705 710 715 720

Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr

725 730 735

Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

740 745 750

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly

755 760 765

Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile

770 775 780

Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly

785 790 795 800

Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser

805 810 815

Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp

820 825 830

Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile

835 840 845

Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu

850 855 860

Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys

865 870 875 880

Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile

885 890 895

Val Met Val Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys

900 905 910

Leu Lys Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp

915 920 925

Asp Ser Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr His His

930 935 940

His His His

945

<210> 52

<211> 901

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 52

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Ile Thr Asn

1 5 10 15

Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val

20 25 30

Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser

35 40 45

Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val

50 55 60

Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp

65 70 75 80

Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln

85 90 95

Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr

100 105 110

Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly

115 120 125

Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys

130 135 140

Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr

145 150 155 160

Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser

165 170 175

Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val

180 185 190

Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly

195 200 205

Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn

210 215 220

Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys

225 230 235 240

Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp

245 250 255

Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys

260 265 270

Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn

275 280 285

Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val

290 295 300

Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr

305 310 315 320

Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu

325 330 335

His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile

340 345 350

Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg Ser

355 360 365

Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu

370 375 380

Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe

385 390 395 400

Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr

405 410 415

Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser

420 425 430

Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala

435 440 445

Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe

450 455 460

Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly

465 470 475 480

Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys

485 490 495

Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp

500 505 510

Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala

515 520 525

Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro

530 535 540

Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu

545 550 555 560

Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu

565 570 575

Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly

580 585 590

Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln

595 600 605

Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala

610 615 620

Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala

625 630 635 640

Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser

645 650 655

Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu

660 665 670

Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr

675 680 685

Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala

690 695 700

Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

705 710 715 720

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln

725 730 735

Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala

740 745 750

Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys

755 760 765

Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His Trp

770 775 780

Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp

785 790 795 800

Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn

805 810 815

Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu

820 825 830

Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu

835 840 845

Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu

850 855 860

Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile

865 870 875 880

Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro His

885 890 895

His His His His His

900

<210> 53

<211> 2706

<212> DNA

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 53

atgttcgtct tcctggtgct gctgcccctg gtgtccagca tcactaacct gtgccccttc 60

ggagaggtct tcaacgccac ccgcttcgct tccgtgtacg cctggaacag gaagagaatc 120

tcaaactgcg tcgctgacta ctccgtgctg tacaactcag cctccttcag caccttcaag 180

tgctacggcg tgtcaccaac taagctgaac gacctgtgct tcaccaacgt ctacgccgac 240

tccttcgtga tcaggggaga cgaggtcaga cagatcgctc ctggccagac tggaaagatc 300

gccgactaca actacaagct gcccgacgac ttcaccggtt gcgtcatcgc ttggaacagc 360

aacaacctgg actctaaagt gggtggcaac tacaactacc tgtaccgcct gttccgtaag 420

tcaaacctga agccattcga gagggacatc agcactgaaa tctaccaggc cggatctacc 480

ccttgcaacg gtgtcgaggg cttcaactgc tacttccccc tgcagtccta cggtttccag 540

ccaactaacg gtgtgggcta ccagccttac agagtggtcg tgctgagctt cgaactgctc 600

cacgctcctg ctactgtgtg cggtccaaag aagtctacca acctggtcaa gaacaagtgc 660

gtgaacttca acttcaacgg cctgaccgga actggtgtcc tgaccgagag caacaagaag 720

ttcctgccct tccagcagtt cggaagggac atcgctgaca ccactgacgc tgtgcgcgac 780

cctcagaccc tggaaatcct ggacatcact ccatgctcat tcggaggtgt ctccgtgatc 840

acccctggca ccaacacttc taaccaggtc gctgtgctgt accaggacgt caactgcacc 900

gaggtccctg tggccatcca cgctgaccag ctgaccccca cttggcgcgt gtactccacc 960

ggcagcaacg tgttccagac tcgtgctggt tgcctgatcg gcgccgagca cgtgaacaac 1020

agctacgaat gcgacatccc catcggcgct ggaatctgcg cctcttacca gacccagact 1080

aacagcccac gcagggctcg ctctgtggcc tctcagtcaa tcatcgctta caccatgtca 1140

ctgggcgctg aaaactccgt ggcctactct aacaactcaa tcgccatccc caccaacttc 1200

actatcagcg tgaccactga gatcctgcca gtcagcatga ccaagacttc tgtggactgc 1260

actatgtaca tctgcggaga cagcaccgaa tgctctaacc tgctgctgca gtacggctct 1320

ttctgcaccc agctgaaccg tgctctgact ggaatcgccg tggagcagga caagaacact 1380

caggaagtct tcgctcaggt gaagcaaatc tacaagaccc cacctatcaa ggacttcggc 1440

ggattcaact tctcccagat cctgcctgac ccctccaagc caagcaagcg ctctttcatc 1500

gaggacctgc tgttcaacaa ggtcactctg gccgacgctg gattcatcaa gcagtacgga 1560

gactgcctgg gtgacatcgc cgctcgtgac ctgatctgcg ctcagaagtt caacggtctg 1620

accgtgctgc ccccactgct gactgacgaa atgatcgccc agtacactag cgccctgctg 1680

gctggaacca tcacttctgg ttggaccttc ggtgctggcg ccgctctgca gatccctttc 1740

gctatgcaga tggcctaccg tttcaacgga atcggtgtca cccagaacgt gctgtacgag 1800

aaccagaagc tgatcgctaa ccagttcaac tcagccatcg gaaagatcca ggacagcctg 1860

agctctactg cctctgctct gggcaagctg caggacgtcg tgaaccagaa cgcccaggct 1920

ctgaacaccc tggtcaagca gctgtcatcc aacttcggtg ctatcagctc tgtgctgaac 1980

gacatcctgt cccgcctgga caaggtcgag gccgaagtgc agatcgaccg cctgatcact 2040

ggccgtctgc agtcactgca gacctacgtg actcagcagc tgatcagggc cgctgaaatc 2100

agagcctccg ctaacctggc cgctaccaag atgagcgagt gcgtcctggg tcaatctaag 2160

cgtgtggact tctgcggcaa gggataccac ctgatgtcat tccctcagtc tgctccccac 2220

ggtgtggtgt tcctgcacgt cacctacgtg ccagcccagg agaagaactt caccactgcc 2280

cctgctatct gccacgacgg caaggctcac ttccccaggg aaggtgtctt cgtgagcaac 2340

ggcacccact ggttcgtcac tcagagaaac ttctacgagc cacagatcat caccactgac 2400

aacactttcg tgtctggaaa ctgcgacgtg gtcatcggta tcgtcaacaa caccgtgtac 2460

gaccccctgc agccagagct ggactcattc aaggaggaac tggacaagta cttcaagaac 2520

cacacctccc ctgacgtcga cctgggcgac atctcaggaa tcaacgcttc cgtcgtgaac 2580

atccagaagg agatcgaccg cctgaacgaa gtggccaaga acctgaacga aagcctgatc 2640

gacctgcagg agctgggcaa gtacgaacag tacatcaagt ggcctcacca ccaccaccac 2700

caccac 2706

<210> 54

<211> 888

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 54

Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe

1 5 10 15

Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala

20 25 30

Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys

35 40 45

Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val

50 55 60

Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala

65 70 75 80

Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp

85 90 95

Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser

100 105 110

Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser

115 120 125

Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala

130 135 140

Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro

145 150 155 160

Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro

165 170 175

Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr

180 185 190

Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val

195 200 205

Asn Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser

210 215 220

Asn Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp

225 230 235 240

Thr Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile

245 250 255

Thr Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn

260 265 270

Thr Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu

275 280 285

Val Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val

290 295 300

Tyr Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile

305 310 315 320

Gly Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly

325 330 335

Ala Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg

340 345 350

Ala Arg Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu

355 360 365

Gly Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro

370 375 380

Thr Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met

385 390 395 400

Thr Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr

405 410 415

Glu Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu

420 425 430

Asn Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln

435 440 445

Glu Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys

450 455 460

Asp Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys

465 470 475 480

Pro Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr

485 490 495

Leu Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp

500 505 510

Ile Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr

515 520 525

Val Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser

530 535 540

Ala Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly

545 550 555 560

Ala Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn

565 570 575

Gly Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile

580 585 590

Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser

595 600 605

Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn

610 615 620

Ala Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly

625 630 635 640

Ala Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val

645 650 655

Glu Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser

660 665 670

Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg

675 680 685

Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly

690 695 700

Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser

705 710 715 720

Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr

725 730 735

Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

740 745 750

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly

755 760 765

Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile

770 775 780

Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly

785 790 795 800

Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser

805 810 815

Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp

820 825 830

Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile

835 840 845

Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu

850 855 860

Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys

865 870 875 880

Trp Pro His His His His His His

885

<210> 55

<211> 3819

<212> DNA

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 55

atgttcgtct tcctggtgct gctgcccctg gtgtccagcc agtgcgtgaa cctgaccact 60

aggactcagc tgcctcccgc ttacaccaac tcattcactc gcggtgtgta ctaccctgac 120

aaggtcttcc gttcttcagt gctgcactca actcaggacc tgttcctgcc cttcttctcc 180

aacgtcacct ggttccacgc catccacgtg tccggcacca acggcactaa gcgcttcgac 240

aacccagtgc tgcctttcaa cgacggtgtc tacttcgctt caaccgagaa gtccaacatc 300

atccgtggat ggatcttcgg caccactctg gacagcaaga ctcagtctct gctgatcgtc 360

aacaacgcca ccaacgtggt catcaaggtc tgcgaattcc agttctgcaa cgacccattc 420

ctgggcgtct actaccacaa gaacaacaag tcatggatgg agtccgaatt ccgcgtctac 480

tccagcgcta acaactgcac tttcgagtac gtgtcccagc ctttcctgat ggacctggaa 540

ggaaagcagg gtaacttcaa gaacctgagg gagttcgtgt tcaagaacat cgacggatac 600

ttcaagattt acagcaagca caccccaatc aacctggtgc gcgacctgcc tcagggtttc 660

tctgctctgg agccactggt ggacctgcct atcggcatca acatcacccg cttccagact 720

ctgctggctc tgcaccgttc ctacctgact ccaggcgact catcttctgg atggactgct 780

ggagctgctg cttactacgt gggctacctg cagcctcgca ccttcctgct gaagtacaac 840

gaaaacggaa ccatcactga cgccgtcgac tgcgctctgg accctctgtc agaaaccaag 900

tgcactctga agtccttcac cgtggagaag ggcatctacc agacttcaaa cttcagggtg 960

cagcccaccg aatccatcgt cagattccct aacatcacta acctgtgccc cttcggagag 1020

gtcttcaacg ccacccgctt cgcttccgtg tacgcctgga acaggaagag aatctcaaac 1080

tgcgtcgctg actactccgt gctgtacaac tcagcctcct tcagcacctt caagtgctac 1140

ggcgtgtcac caactaagct gaacgacctg tgcttcacca acgtctacgc cgactccttc 1200

gtgatcaggg gagacgaggt cagacagatc gctcctggcc agactggaaa gatcgccgac 1260

tacaactaca agctgcccga cgacttcacc ggttgcgtca tcgcttggaa cagcaacaac 1320

ctggactcta aagtgggtgg caactacaac tacctgtacc gcctgttccg taagtcaaac 1380

ctgaagccat tcgagaggga catcagcact gaaatctacc aggccggatc taccccttgc 1440

aacggtgtcg agggcttcaa ctgctacttc cccctgcagt cctacggttt ccagccaact 1500

aacggtgtgg gctaccagcc ttacagagtg gtcgtgctga gcttcgaact gctccacgct 1560

cctgctactg tgtgcggtcc aaagaagtct accaacctgg tcaagaacaa gtgcgtgaac 1620

ttcaacttca acggcctgac cggaactggt gtcctgaccg agagcaacaa gaagttcctg 1680

cccttccagc agttcggaag ggacatcgct gacaccactg acgctgtgcg cgaccctcag 1740

accctggaaa tcctggacat cactccatgc tcattcggag gtgtctccgt gatcacccct 1800

ggcaccaaca cttctaacca ggtcgctgtg ctgtaccagg acgtcaactg caccgaggtc 1860

cctgtggcca tccacgctga ccagctgacc cccacttggc gcgtgtactc caccggcagc 1920

aacgtgttcc agactcgtgc tggttgcctg atcggcgccg agcacgtgaa caacagctac 1980

gaatgcgaca tccccatcgg cgctggaatc tgcgcctctt accagaccca gactaacagc 2040

ccacagcagg ctcagtctgt ggcctctcag tcaatcatcg cttacaccat gtcactgggc 2100

gctgaaaact ccgtggccta ctctaacaac tcaatcgcca tccccaccaa cttcactatc 2160

agcgtgacca ctgagatcct gccagtcagc atgaccaaga cttctgtgga ctgcactatg 2220

tacatctgcg gagacagcac cgaatgctct aacctgctgc tgcagtacgg ctctttctgc 2280

acccagctga accgtgctct gactggaatc gccgtggagc aggacaagaa cactcaggaa 2340

gtcttcgctc aggtgaagca aatctacaag accccaccta tcaaggactt cggcggattc 2400

aacttctccc agatcctgcc tgacccctcc aagccaagca agcgctcttt catcgaggac 2460

ctgctgttca acaaggtcac tctggccgac gctggattca tcaagcagta cggagactgc 2520

ctgggtgaca tcgccgctcg tgacctgatc tgcgctcaga agttcaacgg tctgaccgtg 2580

ctgcccccac tgctgactga cgaaatgatc gcccagtaca ctagcgccct gctggctgga 2640

accatcactt ctggttggac cttcggtgct ggcgccgctc tgcagatccc tttcgctatg 2700

cagatggcct accgtttcaa cggaatcggt gtcacccaga acgtgctgta cgagaaccag 2760

aagctgatcg ctaaccagtt caactcagcc atcggaaaga tccaggacag cctgagctct 2820

actgcctctg ctctgggcaa gctgcaggac gtcgtgaacc agaacgccca ggctctgaac 2880

accctggtca agcagctgtc atccaacttc ggtgctatca gctctgtgct gaacgacatc 2940

ctgtcccgcc tggacaaggt cgaggccgaa gtgcagatcg accgcctgat cactggccgt 3000

ctgcagtcac tgcagaccta cgtgactcag cagctgatca gggccgctga aatcagagcc 3060

tccgctaacc tggccgctac caagatgagc gagtgcgtcc tgggtcaatc taagcgtgtg 3120

gacttctgcg gcaagggata ccacctgatg tcattccctc agtctgctcc ccacggtgtg 3180

gtgttcctgc acgtcaccta cgtgccagcc caggagaaga acttcaccac tgcccctgct 3240

atctgccacg acggcaaggc tcacttcccc agggaaggtg tcttcgtgag caacggcacc 3300

cactggttcg tcactcagag aaacttctac gagccacaga tcatcaccac tgacaacact 3360

ttcgtgtctg gaaactgcga cgtggtcatc ggtatcgtca acaacaccgt gtacgacccc 3420

ctgcagccag agctggactc attcaaggag gaactggaca agtacttcaa gaaccacacc 3480

tcccctgacg tcgacctggg cgacatctca ggaatcaacg cttccgtcgt gaacatccag 3540

aaggagatcg accgcctgaa cgaagtggcc aagaacctga acgaaagcct gatcgacctg 3600

caggagctgg gcaagtacga acagtacatc aagtggcctt ggtacatctg gctgggtttc 3660

atcgctggcc tcatcgctat cgtgatggtg accatcatgc tgtgctgcat gacttcatgc 3720

tgctcctgcc tgaagggctg ctgcagctgc ggatcttgct gcaagttcga cgaggacgac 3780

tctgaacccg tcctgaaggg cgtgaagctg cactacacc 3819

<210> 56

<211> 1279

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 56

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln Ser Val Ala

675 680 685

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser

690 695 700

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

705 710 715 720

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

725 730 735

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

740 745 750

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

755 760 765

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

770 775 780

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe

785 790 795 800

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

805 810 815

Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly

820 825 830

Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp

835 840 845

Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu

850 855 860

Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly

865 870 875 880

Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile

885 890 895

Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr

900 905 910

Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn

915 920 925

Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala

930 935 940

Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn

945 950 955 960

Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val

965 970 975

Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln

980 985 990

Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val

995 1000 1005

Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn

1010 1015 1020

Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

1025 1030 1035

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro

1040 1045 1050

Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val

1055 1060 1065

Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

1070 1075 1080

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

1085 1090 1095

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln

1100 1105 1110

Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val

1115 1120 1125

Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro

1130 1135 1140

Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn

1145 1150 1155

His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn

1160 1165 1170

Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu

1175 1180 1185

Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu

1190 1195 1200

Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu

1205 1210 1215

Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met

1220 1225 1230

Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys

1235 1240 1245

Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro

1250 1255 1260

Val Leu Lys Gly Val Lys Leu His Tyr Thr His His His His His

1265 1270 1275

His

<210> 57

<211> 3837

<212> DNA

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 57

atgttcgtct tcctggtgct gctgcccctg gtgtccagcc agtgcgtgaa cctgaccact 60

aggactcagc tgcctcccgc ttacaccaac tcattcactc gcggtgtgta ctaccctgac 120

aaggtcttcc gttcttcagt gctgcactca actcaggacc tgttcctgcc cttcttctcc 180

aacgtcacct ggttccacgc catccacgtg tccggcacca acggcactaa gcgcttcgac 240

aacccagtgc tgcctttcaa cgacggtgtc tacttcgctt caaccgagaa gtccaacatc 300

atccgtggat ggatcttcgg caccactctg gacagcaaga ctcagtctct gctgatcgtc 360

aacaacgcca ccaacgtggt catcaaggtc tgcgaattcc agttctgcaa cgacccattc 420

ctgggcgtct actaccacaa gaacaacaag tcatggatgg agtccgaatt ccgcgtctac 480

tccagcgcta acaactgcac tttcgagtac gtgtcccagc ctttcctgat ggacctggaa 540

ggaaagcagg gtaacttcaa gaacctgagg gagttcgtgt tcaagaacat cgacggatac 600

ttcaagattt acagcaagca caccccaatc aacctggtgc gcgacctgcc tcagggtttc 660

tctgctctgg agccactggt ggacctgcct atcggcatca acatcacccg cttccagact 720

ctgctggctc tgcaccgttc ctacctgact ccaggcgact catcttctgg atggactgct 780

ggagctgctg cttactacgt gggctacctg cagcctcgca ccttcctgct gaagtacaac 840

gaaaacggaa ccatcactga cgccgtcgac tgcgctctgg accctctgtc agaaaccaag 900

tgcactctga agtccttcac cgtggagaag ggcatctacc agacttcaaa cttcagggtg 960

cagcccaccg aatccatcgt cagattccct aacatcacta acctgtgccc cttcggagag 1020

gtcttcaacg ccacccgctt cgcttccgtg tacgcctgga acaggaagag aatctcaaac 1080

tgcgtcgctg actactccgt gctgtacaac tcagcctcct tcagcacctt caagtgctac 1140

ggcgtgtcac caactaagct gaacgacctg tgcttcacca acgtctacgc cgactccttc 1200

gtgatcaggg gagacgaggt cagacagatc gctcctggcc agactggaaa gatcgccgac 1260

tacaactaca agctgcccga cgacttcacc ggttgcgtca tcgcttggaa cagcaacaac 1320

ctggactcta aagtgggtgg caactacaac tacctgtacc gcctgttccg taagtcaaac 1380

ctgaagccat tcgagaggga catcagcact gaaatctacc aggccggatc taccccttgc 1440

aacggtgtcg agggcttcaa ctgctacttc cccctgcagt cctacggttt ccagccaact 1500

aacggtgtgg gctaccagcc ttacagagtg gtcgtgctga gcttcgaact gctccacgct 1560

cctgctactg tgtgcggtcc aaagaagtct accaacctgg tcaagaacaa gtgcgtgaac 1620

ttcaacttca acggcctgac cggaactggt gtcctgaccg agagcaacaa gaagttcctg 1680

cccttccagc agttcggaag ggacatcgct gacaccactg acgctgtgcg cgaccctcag 1740

accctggaaa tcctggacat cactccatgc tcattcggag gtgtctccgt gatcacccct 1800

ggcaccaaca cttctaacca ggtcgctgtg ctgtaccagg acgtcaactg caccgaggtc 1860

cctgtggcca tccacgctga ccagctgacc cccacttggc gcgtgtactc caccggcagc 1920

aacgtgttcc agactcgtgc tggttgcctg atcggcgccg agcacgtgaa caacagctac 1980

gaatgcgaca tccccatcgg cgctggaatc tgcgcctctt accagaccca gactaacagc 2040

ccacagcagg ctcagtctgt ggcctctcag tcaatcatcg cttacaccat gtcactgggc 2100

gctgaaaact ccgtggccta ctctaacaac tcaatcgcca tccccaccaa cttcactatc 2160

agcgtgacca ctgagatcct gccagtcagc atgaccaaga cttctgtgga ctgcactatg 2220

tacatctgcg gagacagcac cgaatgctct aacctgctgc tgcagtacgg ctctttctgc 2280

acccagctga accgtgctct gactggaatc gccgtggagc aggacaagaa cactcaggaa 2340

gtcttcgctc aggtgaagca aatctacaag accccaccta tcaaggactt cggcggattc 2400

aacttctccc agatcctgcc tgacccctcc aagccaagca agcgctcttt catcgaggac 2460

ctgctgttca acaaggtcac tctggccgac gctggattca tcaagcagta cggagactgc 2520

ctgggtgaca tcgccgctcg tgacctgatc tgcgctcaga agttcaacgg tctgaccgtg 2580

ctgcccccac tgctgactga cgaaatgatc gcccagtaca ctagcgccct gctggctgga 2640

accatcactt ctggttggac cttcggtgct ggcgccgctc tgcagatccc tttcgctatg 2700

cagatggcct accgtttcaa cggaatcggt gtcacccaga acgtgctgta cgagaaccag 2760

aagctgatcg ctaaccagtt caactcagcc atcggaaaga tccaggacag cctgagctct 2820

actgcctctg ctctgggcaa gctgcaggac gtcgtgaacc agaacgccca ggctctgaac 2880

accctggtca agcagctgtc atccaacttc ggtgctatca gctctgtgct gaacgacatc 2940

ctgtcccgcc tggacaaggt cgaggccgaa gtgcagatcg accgcctgat cactggccgt 3000

ctgcagtcac tgcagaccta cgtgactcag cagctgatca gggccgctga aatcagagcc 3060

tccgctaacc tggccgctac caagatgagc gagtgcgtcc tgggtcaatc taagcgtgtg 3120

gacttctgcg gcaagggata ccacctgatg tcattccctc agtctgctcc ccacggtgtg 3180

gtgttcctgc acgtcaccta cgtgccagcc caggagaaga acttcaccac tgcccctgct 3240

atctgccacg acggcaaggc tcacttcccc agggaaggtg tcttcgtgag caacggcacc 3300

cactggttcg tcactcagag aaacttctac gagccacaga tcatcaccac tgacaacact 3360

ttcgtgtctg gaaactgcga cgtggtcatc ggtatcgtca acaacaccgt gtacgacccc 3420

ctgcagccag agctggactc attcaaggag gaactggaca agtacttcaa gaaccacacc 3480

tcccctgacg tcgacctggg cgacatctca ggaatcaacg cttccgtcgt gaacatccag 3540

aaggagatcg accgcctgaa cgaagtggcc aagaacctga acgaaagcct gatcgacctg 3600

caggagctgg gcaagtacga acagtacatc aagtggcctt ggtacatctg gctgggtttc 3660

atcgctggcc tcatcgctat cgtgatggtg accatcatgc tgtgctgcat gacttcatgc 3720

tgctcctgcc tgaagggctg ctgcagctgc ggatcttgct gcaagttcga cgaggacgac 3780

tctgaacccg tcctgaaggg cgtgaagctg cactacaccc accaccacca ccaccac 3837

<210> 58

<211> 1266

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 58

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln

660 665 670

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala

675 680 685

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn

690 695 700

Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys

705 710 715 720

Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys

725 730 735

Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg

740 745 750

Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val

755 760 765

Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe

770 775 780

Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser

785 790 795 800

Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala

805 810 815

Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala

820 825 830

Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu

835 840 845

Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu

850 855 860

Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala

865 870 875 880

Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile

885 890 895

Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn

900 905 910

Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr

915 920 925

Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln

930 935 940

Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile

945 950 955 960

Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala

965 970 975

Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln

980 985 990

Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser

995 1000 1005

Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln

1010 1015 1020

Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser

1025 1030 1035

Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr

1040 1045 1050

Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile

1055 1060 1065

Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val

1070 1075 1080

Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu

1085 1090 1095

Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys

1100 1105 1110

Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu

1115 1120 1125

Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe

1130 1135 1140

Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly

1145 1150 1155

Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu

1160 1165 1170

Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln

1175 1180 1185

Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile

1190 1195 1200

Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr

1205 1210 1215

Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly

1220 1225 1230

Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser

1235 1240 1245

Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr His His His

1250 1255 1260

His His His

1265

<210> 59

<211> 1219

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 59

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln Ser Val Ala

675 680 685

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser

690 695 700

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

705 710 715 720

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

725 730 735

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

740 745 750

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

755 760 765

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

770 775 780

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe

785 790 795 800

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

805 810 815

Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly

820 825 830

Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp

835 840 845

Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu

850 855 860

Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly

865 870 875 880

Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile

885 890 895

Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr

900 905 910

Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn

915 920 925

Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala

930 935 940

Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn

945 950 955 960

Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val

965 970 975

Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln

980 985 990

Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val

995 1000 1005

Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn

1010 1015 1020

Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

1025 1030 1035

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro

1040 1045 1050

Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val

1055 1060 1065

Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

1070 1075 1080

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

1085 1090 1095

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln

1100 1105 1110

Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val

1115 1120 1125

Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro

1130 1135 1140

Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn

1145 1150 1155

His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn

1160 1165 1170

Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu

1175 1180 1185

Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu

1190 1195 1200

Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro His His His His His

1205 1210 1215

His

<210> 60

<211> 3657

<212> DNA

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 60

atgttcgtct tcctggtgct gctgcccctg gtgtccagcc agtgcgtgaa cctgaccact 60

aggactcagc tgcctcccgc ttacaccaac tcattcactc gcggtgtgta ctaccctgac 120

aaggtcttcc gttcttcagt gctgcactca actcaggacc tgttcctgcc cttcttctcc 180

aacgtcacct ggttccacgc catccacgtg tccggcacca acggcactaa gcgcttcgac 240

aacccagtgc tgcctttcaa cgacggtgtc tacttcgctt caaccgagaa gtccaacatc 300

atccgtggat ggatcttcgg caccactctg gacagcaaga ctcagtctct gctgatcgtc 360

aacaacgcca ccaacgtggt catcaaggtc tgcgaattcc agttctgcaa cgacccattc 420

ctgggcgtct actaccacaa gaacaacaag tcatggatgg agtccgaatt ccgcgtctac 480

tccagcgcta acaactgcac tttcgagtac gtgtcccagc ctttcctgat ggacctggaa 540

ggaaagcagg gtaacttcaa gaacctgagg gagttcgtgt tcaagaacat cgacggatac 600

ttcaagattt acagcaagca caccccaatc aacctggtgc gcgacctgcc tcagggtttc 660

tctgctctgg agccactggt ggacctgcct atcggcatca acatcacccg cttccagact 720

ctgctggctc tgcaccgttc ctacctgact ccaggcgact catcttctgg atggactgct 780

ggagctgctg cttactacgt gggctacctg cagcctcgca ccttcctgct gaagtacaac 840

gaaaacggaa ccatcactga cgccgtcgac tgcgctctgg accctctgtc agaaaccaag 900

tgcactctga agtccttcac cgtggagaag ggcatctacc agacttcaaa cttcagggtg 960

cagcccaccg aatccatcgt cagattccct aacatcacta acctgtgccc cttcggagag 1020

gtcttcaacg ccacccgctt cgcttccgtg tacgcctgga acaggaagag aatctcaaac 1080

tgcgtcgctg actactccgt gctgtacaac tcagcctcct tcagcacctt caagtgctac 1140

ggcgtgtcac caactaagct gaacgacctg tgcttcacca acgtctacgc cgactccttc 1200

gtgatcaggg gagacgaggt cagacagatc gctcctggcc agactggaaa gatcgccgac 1260

tacaactaca agctgcccga cgacttcacc ggttgcgtca tcgcttggaa cagcaacaac 1320

ctggactcta aagtgggtgg caactacaac tacctgtacc gcctgttccg taagtcaaac 1380

ctgaagccat tcgagaggga catcagcact gaaatctacc aggccggatc taccccttgc 1440

aacggtgtcg agggcttcaa ctgctacttc cccctgcagt cctacggttt ccagccaact 1500

aacggtgtgg gctaccagcc ttacagagtg gtcgtgctga gcttcgaact gctccacgct 1560

cctgctactg tgtgcggtcc aaagaagtct accaacctgg tcaagaacaa gtgcgtgaac 1620

ttcaacttca acggcctgac cggaactggt gtcctgaccg agagcaacaa gaagttcctg 1680

cccttccagc agttcggaag ggacatcgct gacaccactg acgctgtgcg cgaccctcag 1740

accctggaaa tcctggacat cactccatgc tcattcggag gtgtctccgt gatcacccct 1800

ggcaccaaca cttctaacca ggtcgctgtg ctgtaccagg acgtcaactg caccgaggtc 1860

cctgtggcca tccacgctga ccagctgacc cccacttggc gcgtgtactc caccggcagc 1920

aacgtgttcc agactcgtgc tggttgcctg atcggcgccg agcacgtgaa caacagctac 1980

gaatgcgaca tccccatcgg cgctggaatc tgcgcctctt accagaccca gactaacagc 2040

ccacagcagg ctcagtctgt ggcctctcag tcaatcatcg cttacaccat gtcactgggc 2100

gctgaaaact ccgtggccta ctctaacaac tcaatcgcca tccccaccaa cttcactatc 2160

agcgtgacca ctgagatcct gccagtcagc atgaccaaga cttctgtgga ctgcactatg 2220

tacatctgcg gagacagcac cgaatgctct aacctgctgc tgcagtacgg ctctttctgc 2280

acccagctga accgtgctct gactggaatc gccgtggagc aggacaagaa cactcaggaa 2340

gtcttcgctc aggtgaagca aatctacaag accccaccta tcaaggactt cggcggattc 2400

aacttctccc agatcctgcc tgacccctcc aagccaagca agcgctcttt catcgaggac 2460

ctgctgttca acaaggtcac tctggccgac gctggattca tcaagcagta cggagactgc 2520

ctgggtgaca tcgccgctcg tgacctgatc tgcgctcaga agttcaacgg tctgaccgtg 2580

ctgcccccac tgctgactga cgaaatgatc gcccagtaca ctagcgccct gctggctgga 2640

accatcactt ctggttggac cttcggtgct ggcgccgctc tgcagatccc tttcgctatg 2700

cagatggcct accgtttcaa cggaatcggt gtcacccaga acgtgctgta cgagaaccag 2760

aagctgatcg ctaaccagtt caactcagcc atcggaaaga tccaggacag cctgagctct 2820

actgcctctg ctctgggcaa gctgcaggac gtcgtgaacc agaacgccca ggctctgaac 2880

accctggtca agcagctgtc atccaacttc ggtgctatca gctctgtgct gaacgacatc 2940

ctgtcccgcc tggacaaggt cgaggccgaa gtgcagatcg accgcctgat cactggccgt 3000

ctgcagtcac tgcagaccta cgtgactcag cagctgatca gggccgctga aatcagagcc 3060

tccgctaacc tggccgctac caagatgagc gagtgcgtcc tgggtcaatc taagcgtgtg 3120

gacttctgcg gcaagggata ccacctgatg tcattccctc agtctgctcc ccacggtgtg 3180

gtgttcctgc acgtcaccta cgtgccagcc caggagaaga acttcaccac tgcccctgct 3240

atctgccacg acggcaaggc tcacttcccc agggaaggtg tcttcgtgag caacggcacc 3300

cactggttcg tcactcagag aaacttctac gagccacaga tcatcaccac tgacaacact 3360

ttcgtgtctg gaaactgcga cgtggtcatc ggtatcgtca acaacaccgt gtacgacccc 3420

ctgcagccag agctggactc attcaaggag gaactggaca agtacttcaa gaaccacacc 3480

tcccctgacg tcgacctggg cgacatctca ggaatcaacg cttccgtcgt gaacatccag 3540

aaggagatcg accgcctgaa cgaagtggcc aagaacctga acgaaagcct gatcgacctg 3600

caggagctgg gcaagtacga acagtacatc aagtggcctc accaccacca ccaccac 3657

<210> 61

<211> 1205

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 61

Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn

1 5 10 15

Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser

20 25 30

Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val

35 40 45

Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg

50 55 60

Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser

65 70 75 80

Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu

85 90 95

Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val

100 105 110

Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly

115 120 125

Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg

130 135 140

Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro

145 150 155 160

Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg

165 170 175

Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys

180 185 190

His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala

195 200 205

Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe

210 215 220

Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser

225 230 235 240

Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu

245 250 255

Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr

260 265 270

Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr

275 280 285

Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe

290 295 300

Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn

305 310 315 320

Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val

325 330 335

Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser

340 345 350

Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val

355 360 365

Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp

370 375 380

Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln

385 390 395 400

Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr

405 410 415

Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly

420 425 430

Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys

435 440 445

Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr

450 455 460

Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser

465 470 475 480

Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val

485 490 495

Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly

500 505 510

Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn

515 520 525

Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys

530 535 540

Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp

545 550 555 560

Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys

565 570 575

Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn

580 585 590

Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val

595 600 605

Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr

610 615 620

Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu

625 630 635 640

His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile

645 650 655

Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln Ser

660 665 670

Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu

675 680 685

Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe

690 695 700

Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr

705 710 715 720

Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser

725 730 735

Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala

740 745 750

Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe

755 760 765

Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly

770 775 780

Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys

785 790 795 800

Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp

805 810 815

Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala

820 825 830

Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro

835 840 845

Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu

850 855 860

Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu

865 870 875 880

Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly

885 890 895

Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln

900 905 910

Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala

915 920 925

Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala

930 935 940

Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser

945 950 955 960

Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu

965 970 975

Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr

980 985 990

Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala

995 1000 1005

Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser

1010 1015 1020

Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe

1025 1030 1035

Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr

1040 1045 1050

Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys

1055 1060 1065

His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser

1070 1075 1080

Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro

1085 1090 1095

Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp

1100 1105 1110

Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln

1115 1120 1125

Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys

1130 1135 1140

Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile

1145 1150 1155

Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn

1160 1165 1170

Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu

1175 1180 1185

Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro His His His His

1190 1195 1200

His His

1205

<210> 62

<211> 1263

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 62

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln Ser Val Ala

675 680 685

Leu Gly Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile

690 695 700

Pro Thr Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser

705 710 715 720

Met Thr Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser

725 730 735

Thr Glu Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln

740 745 750

Leu Asn Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr

755 760 765

Gln Glu Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile

770 775 780

Lys Asp Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser

785 790 795 800

Lys Pro Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val

805 810 815

Thr Leu Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly

820 825 830

Asp Ile Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu

835 840 845

Thr Val Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr

850 855 860

Ser Ala Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala

865 870 875 880

Gly Ala Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe

885 890 895

Asn Gly Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu

900 905 910

Ile Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu

915 920 925

Ser Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln

930 935 940

Asn Ala Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe

945 950 955 960

Gly Ala Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys

965 970 975

Val Glu Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln

980 985 990

Ser Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile

995 1000 1005

Arg Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val

1010 1015 1020

Leu Gly Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His

1025 1030 1035

Leu Met Ser Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu

1040 1045 1050

His Val Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala

1055 1060 1065

Pro Ala Ile Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly

1070 1075 1080

Val Phe Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn

1085 1090 1095

Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser

1100 1105 1110

Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr

1115 1120 1125

Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp

1130 1135 1140

Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp

1145 1150 1155

Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile

1160 1165 1170

Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile

1175 1180 1185

Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro

1190 1195 1200

Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val

1205 1210 1215

Met Val Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys

1220 1225 1230

Leu Lys Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu

1235 1240 1245

Asp Asp Ser Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255 1260

<210> 63

<211> 1250

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 63

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln

660 665 670

Ser Val Ala Leu Gly Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser

675 680 685

Ile Ala Ile Pro Thr Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu

690 695 700

Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys

705 710 715 720

Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe

725 730 735

Cys Thr Gln Leu Asn Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp

740 745 750

Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr

755 760 765

Pro Pro Ile Lys Asp Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro

770 775 780

Asp Pro Ser Lys Pro Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe

785 790 795 800

Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp

805 810 815

Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe

820 825 830

Asn Gly Leu Thr Val Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala

835 840 845

Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr

850 855 860

Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala

865 870 875 880

Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn

885 890 895

Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln

900 905 910

Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val

915 920 925

Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser

930 935 940

Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg

945 950 955 960

Leu Asp Lys Val Glu Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly

965 970 975

Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala

980 985 990

Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu

995 1000 1005

Cys Val Leu Gly Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly

1010 1015 1020

Tyr His Leu Met Ser Phe Pro Gln Ser Ala Pro His Gly Val Val

1025 1030 1035

Phe Leu His Val Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr

1040 1045 1050

Thr Ala Pro Ala Ile Cys His Asp Gly Lys Ala His Phe Pro Arg

1055 1060 1065

Glu Gly Val Phe Val Ser Asn Gly Thr His Trp Phe Val Thr Gln

1070 1075 1080

Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe

1085 1090 1095

Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr

1100 1105 1110

Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu

1115 1120 1125

Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu

1130 1135 1140

Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys

1145 1150 1155

Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser

1160 1165 1170

Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys

1175 1180 1185

Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala

1190 1195 1200

Ile Val Met Val Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys

1205 1210 1215

Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe

1220 1225 1230

Asp Glu Asp Asp Ser Glu Pro Val Leu Lys Gly Val Lys Leu His

1235 1240 1245

Tyr Thr

1250

<210> 64

<211> 1263

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 64

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln Ser Val Ala

675 680 685

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser

690 695 700

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Glu Ile Leu Pro Val Ser

705 710 715 720

Met Thr Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser

725 730 735

Thr Glu Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln

740 745 750

Leu Asn Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr

755 760 765

Gln Glu Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile

770 775 780

Lys Asp Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser

785 790 795 800

Lys Pro Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val

805 810 815

Thr Leu Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly

820 825 830

Asp Ile Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu

835 840 845

Thr Val Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr

850 855 860

Ser Ala Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala

865 870 875 880

Gly Ala Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe

885 890 895

Asn Gly Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu

900 905 910

Ile Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu

915 920 925

Ser Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln

930 935 940

Asn Ala Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe

945 950 955 960

Gly Ala Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys

965 970 975

Val Glu Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln

980 985 990

Ser Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile

995 1000 1005

Arg Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val

1010 1015 1020

Leu Gly Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His

1025 1030 1035

Leu Met Ser Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu

1040 1045 1050

His Val Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala

1055 1060 1065

Pro Ala Ile Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly

1070 1075 1080

Val Phe Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn

1085 1090 1095

Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser

1100 1105 1110

Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr

1115 1120 1125

Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp

1130 1135 1140

Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp

1145 1150 1155

Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile

1160 1165 1170

Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile

1175 1180 1185

Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro

1190 1195 1200

Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val

1205 1210 1215

Met Val Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys

1220 1225 1230

Leu Lys Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu

1235 1240 1245

Asp Asp Ser Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255 1260

<210> 65

<211> 1250

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 65

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln

660 665 670

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala

675 680 685

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Glu Ile Leu

690 695 700

Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys

705 710 715 720

Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe

725 730 735

Cys Thr Gln Leu Asn Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp

740 745 750

Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr

755 760 765

Pro Pro Ile Lys Asp Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro

770 775 780

Asp Pro Ser Lys Pro Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe

785 790 795 800

Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp

805 810 815

Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe

820 825 830

Asn Gly Leu Thr Val Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala

835 840 845

Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr

850 855 860

Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala

865 870 875 880

Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn

885 890 895

Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln

900 905 910

Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val

915 920 925

Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser

930 935 940

Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg

945 950 955 960

Leu Asp Lys Val Glu Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly

965 970 975

Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala

980 985 990

Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu

995 1000 1005

Cys Val Leu Gly Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly

1010 1015 1020

Tyr His Leu Met Ser Phe Pro Gln Ser Ala Pro His Gly Val Val

1025 1030 1035

Phe Leu His Val Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr

1040 1045 1050

Thr Ala Pro Ala Ile Cys His Asp Gly Lys Ala His Phe Pro Arg

1055 1060 1065

Glu Gly Val Phe Val Ser Asn Gly Thr His Trp Phe Val Thr Gln

1070 1075 1080

Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe

1085 1090 1095

Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr

1100 1105 1110

Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu

1115 1120 1125

Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu

1130 1135 1140

Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys

1145 1150 1155

Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser

1160 1165 1170

Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys

1175 1180 1185

Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala

1190 1195 1200

Ile Val Met Val Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys

1205 1210 1215

Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe

1220 1225 1230

Asp Glu Asp Asp Ser Glu Pro Val Leu Lys Gly Val Lys Leu His

1235 1240 1245

Tyr Thr

1250

<210> 66

<211> 1263

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 66

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln Ser Val Ala

675 680 685

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser

690 695 700

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

705 710 715 720

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

725 730 735

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

740 745 750

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

755 760 765

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

770 775 780

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe

785 790 795 800

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

805 810 815

Phe Ile Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly

820 825 830

Asp Ile Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu

835 840 845

Thr Val Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr

850 855 860

Ser Ala Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala

865 870 875 880

Gly Ala Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe

885 890 895

Asn Gly Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu

900 905 910

Ile Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu

915 920 925

Ser Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln

930 935 940

Asn Ala Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe

945 950 955 960

Gly Ala Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys

965 970 975

Val Glu Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln

980 985 990

Ser Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile

995 1000 1005

Arg Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val

1010 1015 1020

Leu Gly Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His

1025 1030 1035

Leu Met Ser Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu

1040 1045 1050

His Val Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala

1055 1060 1065

Pro Ala Ile Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly

1070 1075 1080

Val Phe Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn

1085 1090 1095

Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser

1100 1105 1110

Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr

1115 1120 1125

Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp

1130 1135 1140

Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp

1145 1150 1155

Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile

1160 1165 1170

Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile

1175 1180 1185

Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro

1190 1195 1200

Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val

1205 1210 1215

Met Val Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys

1220 1225 1230

Leu Lys Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu

1235 1240 1245

Asp Asp Ser Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255 1260

<210> 67

<211> 1250

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 67

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln

660 665 670

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala

675 680 685

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn

690 695 700

Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys

705 710 715 720

Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys

725 730 735

Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg

740 745 750

Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val

755 760 765

Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe

770 775 780

Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser

785 790 795 800

Lys Arg Ser Phe Ile Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp

805 810 815

Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe

820 825 830

Asn Gly Leu Thr Val Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala

835 840 845

Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr

850 855 860

Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala

865 870 875 880

Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn

885 890 895

Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln

900 905 910

Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val

915 920 925

Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser

930 935 940

Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg

945 950 955 960

Leu Asp Lys Val Glu Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly

965 970 975

Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala

980 985 990

Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu

995 1000 1005

Cys Val Leu Gly Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly

1010 1015 1020

Tyr His Leu Met Ser Phe Pro Gln Ser Ala Pro His Gly Val Val

1025 1030 1035

Phe Leu His Val Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr

1040 1045 1050

Thr Ala Pro Ala Ile Cys His Asp Gly Lys Ala His Phe Pro Arg

1055 1060 1065

Glu Gly Val Phe Val Ser Asn Gly Thr His Trp Phe Val Thr Gln

1070 1075 1080

Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe

1085 1090 1095

Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr

1100 1105 1110

Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu

1115 1120 1125

Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu

1130 1135 1140

Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys

1145 1150 1155

Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser

1160 1165 1170

Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys

1175 1180 1185

Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala

1190 1195 1200

Ile Val Met Val Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys

1205 1210 1215

Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe

1220 1225 1230

Asp Glu Asp Asp Ser Glu Pro Val Leu Lys Gly Val Lys Leu His

1235 1240 1245

Tyr Thr

1250

<210> 68

<211> 31

<212> PRT

<213> 人工序列

<220>

<223> 折叠子

<400> 68

Ser Ala Ile Gly Gly Tyr Ile Pro Glu Ala Pro Arg Asp Gly Gln Ala

1 5 10 15

Tyr Val Arg Lys Asp Gly Glu Trp Val Leu Leu Ser Thr Phe Leu

20 25 30

<210> 69

<211> 202

<212> PRT

<213> β冠状病毒严重急性呼吸系统综合征冠状病毒 2

<400> 69

Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn

1 5 10 15

Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser

20 25 30

Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser

35 40 45

Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys

50 55 60

Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val

65 70 75 80

Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr

85 90 95

Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn

100 105 110

Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu

115 120 125

Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu

130 135 140

Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn

145 150 155 160

Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val

165 170 175

Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His

180 185 190

Ala Pro Ala Thr Val Cys Gly Pro Lys Lys

195 200

<210> 70

<211> 201

<212> PRT

<213> β冠状病毒严重急性呼吸系统综合征相关的冠状病毒

<400> 70

Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn

1 5 10 15

Ala Thr Lys Phe Pro Ser Val Tyr Ala Trp Glu Arg Lys Lys Ile Ser

20 25 30

Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Thr Phe Phe Ser

35 40 45

Thr Phe Lys Cys Tyr Gly Val Ser Ala Thr Lys Leu Asn Asp Leu Cys

50 55 60

Phe Ser Asn Val Tyr Ala Asp Ser Phe Val Val Lys Gly Asp Asp Val

65 70 75 80

Arg Gln Ile Ala Pro Gly Gln Thr Gly Val Ile Ala Asp Tyr Asn Tyr

85 90 95

Lys Leu Pro Asp Asp Phe Met Gly Cys Val Leu Ala Trp Asn Thr Arg

100 105 110

Asn Ile Asp Ala Thr Ser Thr Gly Asn Tyr Asn Tyr Lys Tyr Arg Tyr

115 120 125

Leu Arg His Gly Lys Leu Arg Pro Phe Glu Arg Asp Ile Ser Asn Val

130 135 140

Pro Phe Ser Pro Asp Gly Lys Pro Cys Thr Pro Pro Ala Leu Asn Cys

145 150 155 160

Tyr Trp Pro Leu Asn Asp Tyr Gly Phe Tyr Thr Thr Thr Gly Ile Gly

165 170 175

Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu Asn Ala

180 185 190

Pro Ala Thr Val Cys Gly Pro Lys Leu

195 200

<210> 71

<211> 239

<212> PRT

<213> β冠状病毒中东呼吸系统综合征相关冠状病毒

<400> 71

Phe Glu Ala Lys Pro Ser Gly Ser Val Val Ala Glu Gly Val Glu Cys

1 5 10 15

Asp Phe Ser Pro Leu Leu Ser Gly Thr Pro Pro Gln Val Tyr Asn Phe

20 25 30

Lys Arg Leu Val Phe Thr Asn Cys Asn Tyr Asn Leu Thr Lys Leu Leu

35 40 45

Ser Leu Phe Ser Val Asn Asp Phe Thr Cys Ser Gln Ile Ser Pro Ala

50 55 60

Ala Ile Ala Ser Asn Cys Tyr Ser Ser Leu Ile Leu Asp Tyr Phe Ser

65 70 75 80

Tyr Pro Leu Ser Met Lys Ser Asp Leu Ser Val Ser Ser Ala Gly Pro

85 90 95

Ile Ser Gln Phe Asn Tyr Lys Gln Ser Phe Ser Asn Pro Thr Cys Leu

100 105 110

Ile Leu Ala Thr Val Pro His Asn Leu Thr Thr Ile Thr Lys Pro Leu

115 120 125

Lys Tyr Ser Tyr Ile Asn Lys Cys Ser Arg Leu Leu Ser Asp Asp Arg

130 135 140

Thr Glu Val Pro Gln Leu Val Asn Ala Asn Gln Tyr Ser Pro Cys Val

145 150 155 160

Ser Ile Val Pro Ser Thr Val Trp Glu Asp Gly Asp Tyr Tyr Arg Lys

165 170 175

Gln Leu Ser Pro Leu Glu Gly Gly Gly Trp Leu Val Ala Ser Gly Ser

180 185 190

Thr Val Ala Met Thr Glu Gln Leu Gln Met Gly Phe Gly Ile Thr Val

195 200 205

Gln Tyr Gly Thr Asp Thr Asn Ser Val Cys Pro Lys Leu Glu Phe Ala

210 215 220

Asn Asp Thr Lys Ile Ala Ser Gln Leu Gly Asn Cys Val Glu Tyr

225 230 235

<210> 72

<211> 544

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 72

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Arg Phe Pro

1 5 10 15

Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg

20 25 30

Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val

35 40 45

Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys

50 55 60

Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn

65 70 75 80

Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile

85 90 95

Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro

100 105 110

Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp

115 120 125

Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys

130 135 140

Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln

145 150 155 160

Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe

165 170 175

Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln

180 185 190

Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala

195 200 205

Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys

210 215 220

Val Asn Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu

225 230 235 240

Ser Asn Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala

245 250 255

Asp Thr Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp

260 265 270

Ile Thr Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr

275 280 285

Asn Thr Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr

290 295 300

Glu Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe

305 310 315 320

Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile

325 330 335

Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe

340 345 350

Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu

355 360 365

Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu

370 375 380

Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn

385 390 395 400

Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser

405 410 415

Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg

420 425 430

Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr

435 440 445

Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe

450 455 460

Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly

465 470 475 480

Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu

485 490 495

His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Ala Ile Gly Gly

500 505 510

Tyr Ile Pro Glu Ala Pro Arg Asp Gly Gln Ala Tyr Val Arg Lys Asp

515 520 525

Gly Glu Trp Val Leu Leu Ser Thr Phe Leu His His His His His His

530 535 540

<210> 73

<211> 531

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 73

Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn

1 5 10 15

Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser

20 25 30

Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser

35 40 45

Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys

50 55 60

Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val

65 70 75 80

Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr

85 90 95

Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn

100 105 110

Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu

115 120 125

Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu

130 135 140

Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn

145 150 155 160

Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val

165 170 175

Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His

180 185 190

Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys

195 200 205

Asn Lys Cys Val Asn Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val

210 215 220

Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg

225 230 235 240

Asp Ile Ala Asp Thr Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu

245 250 255

Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr

260 265 270

Pro Gly Thr Asn Thr Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val

275 280 285

Asn Cys Thr Glu Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly

290 295 300

Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg

305 310 315 320

Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser

325 330 335

Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu

340 345 350

Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg

355 360 365

Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala

370 375 380

Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala

385 390 395 400

Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr

405 410 415

Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp

420 425 430

Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val

435 440 445

Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro

450 455 460

Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe

465 470 475 480

Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Ala

485 490 495

Ile Gly Gly Tyr Ile Pro Glu Ala Pro Arg Asp Gly Gln Ala Tyr Val

500 505 510

Arg Lys Asp Gly Glu Trp Val Leu Leu Ser Thr Phe Leu His His His

515 520 525

His His His

530

<210> 74

<211> 544

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 74

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Arg Phe Pro

1 5 10 15

Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg

20 25 30

Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val

35 40 45

Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys

50 55 60

Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn

65 70 75 80

Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile

85 90 95

Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro

100 105 110

Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp

115 120 125

Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys

130 135 140

Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln

145 150 155 160

Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe

165 170 175

Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln

180 185 190

Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala

195 200 205

Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys

210 215 220

Val Asn Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu

225 230 235 240

Ser Asn Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala

245 250 255

Asp Thr Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp

260 265 270

Ile Thr Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr

275 280 285

Asn Thr Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr

290 295 300

Glu Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe

305 310 315 320

Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile

325 330 335

Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe

340 345 350

Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu

355 360 365

Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu

370 375 380

Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn

385 390 395 400

Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser

405 410 415

Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg

420 425 430

Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr

435 440 445

Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe

450 455 460

Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly

465 470 475 480

Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu

485 490 495

His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Ala Ile Gly Gly

500 505 510

Tyr Ile Pro Glu Ala Pro Arg Asp Gly Gln Ala Tyr Val Arg Lys Asp

515 520 525

Gly Glu Trp Val Leu Leu Ser Thr Phe Leu His His His His His His

530 535 540

<210> 75

<211> 531

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 75

Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn

1 5 10 15

Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser

20 25 30

Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser

35 40 45

Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys

50 55 60

Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val

65 70 75 80

Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr

85 90 95

Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn

100 105 110

Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu

115 120 125

Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu

130 135 140

Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn

145 150 155 160

Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val

165 170 175

Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His

180 185 190

Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys

195 200 205

Asn Lys Cys Val Asn Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val

210 215 220

Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg

225 230 235 240

Asp Ile Ala Asp Thr Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu

245 250 255

Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr

260 265 270

Pro Gly Thr Asn Thr Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val

275 280 285

Asn Cys Thr Glu Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly

290 295 300

Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg

305 310 315 320

Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser

325 330 335

Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu

340 345 350

Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg

355 360 365

Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala

370 375 380

Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala

385 390 395 400

Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr

405 410 415

Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp

420 425 430

Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val

435 440 445

Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro

450 455 460

Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe

465 470 475 480

Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Ala

485 490 495

Ile Gly Gly Tyr Ile Pro Glu Ala Pro Arg Asp Gly Gln Ala Tyr Val

500 505 510

Arg Lys Asp Gly Glu Trp Val Leu Leu Ser Thr Phe Leu His His His

515 520 525

His His His

530

<210> 76

<211> 470

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 76

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Arg Phe Pro

1 5 10 15

Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg

20 25 30

Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val

35 40 45

Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys

50 55 60

Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn

65 70 75 80

Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile

85 90 95

Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro

100 105 110

Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp

115 120 125

Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys

130 135 140

Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln

145 150 155 160

Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe

165 170 175

Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln

180 185 190

Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala

195 200 205

Thr Val Cys Gly Pro Lys Lys Ser Gly Gly Gly Ser Gly Gly Gly Ser

210 215 220

Gly Gly Gly Ser Gly Gly Gly Ser Arg Phe Pro Asn Ile Thr Asn Leu

225 230 235 240

Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Lys Phe Pro Ser Val Tyr

245 250 255

Ala Trp Glu Arg Lys Lys Ile Ser Asn Cys Val Ala Asp Tyr Ser Val

260 265 270

Leu Tyr Asn Ser Thr Phe Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser

275 280 285

Ala Thr Lys Leu Asn Asp Leu Cys Phe Ser Asn Val Tyr Ala Asp Ser

290 295 300

Phe Val Val Lys Gly Asp Asp Val Arg Gln Ile Ala Pro Gly Gln Thr

305 310 315 320

Gly Val Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Met Gly

325 330 335

Cys Val Leu Ala Trp Asn Thr Arg Asn Ile Asp Ala Thr Ser Thr Gly

340 345 350

Asn Tyr Asn Tyr Lys Tyr Arg Tyr Leu Arg His Gly Lys Leu Arg Pro

355 360 365

Phe Glu Arg Asp Ile Ser Asn Val Pro Phe Ser Pro Asp Gly Lys Pro

370 375 380

Cys Thr Pro Pro Ala Leu Asn Cys Tyr Trp Pro Leu Asn Asp Tyr Gly

385 390 395 400

Phe Tyr Thr Thr Thr Gly Ile Gly Tyr Gln Pro Tyr Arg Val Val Val

405 410 415

Leu Ser Phe Glu Leu Leu Asn Ala Pro Ala Thr Val Cys Gly Pro Lys

420 425 430

Leu Ser Ala Ile Gly Gly Tyr Ile Pro Glu Ala Pro Arg Asp Gly Gln

435 440 445

Ala Tyr Val Arg Lys Asp Gly Glu Trp Val Leu Leu Ser Thr Phe Leu

450 455 460

His His His His His His

465 470

<210> 77

<211> 464

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 77

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Arg Phe Pro

1 5 10 15

Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg

20 25 30

Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val

35 40 45

Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys

50 55 60

Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn

65 70 75 80

Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile

85 90 95

Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro

100 105 110

Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp

115 120 125

Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys

130 135 140

Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln

145 150 155 160

Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe

165 170 175

Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln

180 185 190

Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala

195 200 205

Thr Val Cys Gly Pro Lys Lys Ser Gly Gly Gly Ser Gly Gly Gly Ser

210 215 220

Gly Gly Gly Ser Gly Gly Gly Ser Arg Phe Pro Asn Ile Thr Asn Leu

225 230 235 240

Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Lys Phe Pro Ser Val Tyr

245 250 255

Ala Trp Glu Arg Lys Lys Ile Ser Asn Cys Val Ala Asp Tyr Ser Val

260 265 270

Leu Tyr Asn Ser Thr Phe Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser

275 280 285

Ala Thr Lys Leu Asn Asp Leu Cys Phe Ser Asn Val Tyr Ala Asp Ser

290 295 300

Phe Val Val Lys Gly Asp Asp Val Arg Gln Ile Ala Pro Gly Gln Thr

305 310 315 320

Gly Val Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Met Gly

325 330 335

Cys Val Leu Ala Trp Asn Thr Arg Asn Ile Asp Ala Thr Ser Thr Gly

340 345 350

Asn Tyr Asn Tyr Lys Tyr Arg Tyr Leu Arg His Gly Lys Leu Arg Pro

355 360 365

Phe Glu Arg Asp Ile Ser Asn Val Pro Phe Ser Pro Asp Gly Lys Pro

370 375 380

Cys Thr Pro Pro Ala Leu Asn Cys Tyr Trp Pro Leu Asn Asp Tyr Gly

385 390 395 400

Phe Tyr Thr Thr Thr Gly Ile Gly Tyr Gln Pro Tyr Arg Val Val Val

405 410 415

Leu Ser Phe Glu Leu Leu Asn Ala Pro Ala Thr Val Cys Gly Pro Lys

420 425 430

Leu Ser Ala Ile Gly Gly Tyr Ile Pro Glu Ala Pro Arg Asp Gly Gln

435 440 445

Ala Tyr Val Arg Lys Asp Gly Glu Trp Val Leu Leu Ser Thr Phe Leu

450 455 460

<210> 78

<211> 457

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 78

Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn

1 5 10 15

Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser

20 25 30

Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser

35 40 45

Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys

50 55 60

Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val

65 70 75 80

Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr

85 90 95

Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn

100 105 110

Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu

115 120 125

Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu

130 135 140

Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn

145 150 155 160

Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val

165 170 175

Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His

180 185 190

Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Gly Gly Gly Ser Gly

195 200 205

Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Phe Pro Asn Ile

210 215 220

Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Lys Phe Pro

225 230 235 240

Ser Val Tyr Ala Trp Glu Arg Lys Lys Ile Ser Asn Cys Val Ala Asp

245 250 255

Tyr Ser Val Leu Tyr Asn Ser Thr Phe Phe Ser Thr Phe Lys Cys Tyr

260 265 270

Gly Val Ser Ala Thr Lys Leu Asn Asp Leu Cys Phe Ser Asn Val Tyr

275 280 285

Ala Asp Ser Phe Val Val Lys Gly Asp Asp Val Arg Gln Ile Ala Pro

290 295 300

Gly Gln Thr Gly Val Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp

305 310 315 320

Phe Met Gly Cys Val Leu Ala Trp Asn Thr Arg Asn Ile Asp Ala Thr

325 330 335

Ser Thr Gly Asn Tyr Asn Tyr Lys Tyr Arg Tyr Leu Arg His Gly Lys

340 345 350

Leu Arg Pro Phe Glu Arg Asp Ile Ser Asn Val Pro Phe Ser Pro Asp

355 360 365

Gly Lys Pro Cys Thr Pro Pro Ala Leu Asn Cys Tyr Trp Pro Leu Asn

370 375 380

Asp Tyr Gly Phe Tyr Thr Thr Thr Gly Ile Gly Tyr Gln Pro Tyr Arg

385 390 395 400

Val Val Val Leu Ser Phe Glu Leu Leu Asn Ala Pro Ala Thr Val Cys

405 410 415

Gly Pro Lys Leu Ser Ala Ile Gly Gly Tyr Ile Pro Glu Ala Pro Arg

420 425 430

Asp Gly Gln Ala Tyr Val Arg Lys Asp Gly Glu Trp Val Leu Leu Ser

435 440 445

Thr Phe Leu His His His His His His

450 455

<210> 79

<211> 451

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 79

Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn

1 5 10 15

Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser

20 25 30

Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser

35 40 45

Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys

50 55 60

Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val

65 70 75 80

Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr

85 90 95

Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn

100 105 110

Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu

115 120 125

Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu

130 135 140

Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn

145 150 155 160

Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val

165 170 175

Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His

180 185 190

Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Gly Gly Gly Ser Gly

195 200 205

Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Phe Pro Asn Ile

210 215 220

Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Lys Phe Pro

225 230 235 240

Ser Val Tyr Ala Trp Glu Arg Lys Lys Ile Ser Asn Cys Val Ala Asp

245 250 255

Tyr Ser Val Leu Tyr Asn Ser Thr Phe Phe Ser Thr Phe Lys Cys Tyr

260 265 270

Gly Val Ser Ala Thr Lys Leu Asn Asp Leu Cys Phe Ser Asn Val Tyr

275 280 285

Ala Asp Ser Phe Val Val Lys Gly Asp Asp Val Arg Gln Ile Ala Pro

290 295 300

Gly Gln Thr Gly Val Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp

305 310 315 320

Phe Met Gly Cys Val Leu Ala Trp Asn Thr Arg Asn Ile Asp Ala Thr

325 330 335

Ser Thr Gly Asn Tyr Asn Tyr Lys Tyr Arg Tyr Leu Arg His Gly Lys

340 345 350

Leu Arg Pro Phe Glu Arg Asp Ile Ser Asn Val Pro Phe Ser Pro Asp

355 360 365

Gly Lys Pro Cys Thr Pro Pro Ala Leu Asn Cys Tyr Trp Pro Leu Asn

370 375 380

Asp Tyr Gly Phe Tyr Thr Thr Thr Gly Ile Gly Tyr Gln Pro Tyr Arg

385 390 395 400

Val Val Val Leu Ser Phe Glu Leu Leu Asn Ala Pro Ala Thr Val Cys

405 410 415

Gly Pro Lys Leu Ser Ala Ile Gly Gly Tyr Ile Pro Glu Ala Pro Arg

420 425 430

Asp Gly Gln Ala Tyr Val Arg Lys Asp Gly Glu Trp Val Leu Leu Ser

435 440 445

Thr Phe Leu

450

<210> 80

<211> 726

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 80

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Arg Phe Pro

1 5 10 15

Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg

20 25 30

Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val

35 40 45

Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys

50 55 60

Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn

65 70 75 80

Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile

85 90 95

Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro

100 105 110

Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp

115 120 125

Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys

130 135 140

Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln

145 150 155 160

Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe

165 170 175

Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln

180 185 190

Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala

195 200 205

Thr Val Cys Gly Pro Lys Lys Ser Gly Gly Gly Ser Gly Gly Gly Ser

210 215 220

Gly Gly Gly Ser Gly Gly Gly Ser Phe Glu Ala Lys Pro Ser Gly Ser

225 230 235 240

Val Val Ala Glu Gly Val Glu Cys Asp Phe Ser Pro Leu Leu Ser Gly

245 250 255

Thr Pro Pro Gln Val Tyr Asn Phe Lys Arg Leu Val Phe Thr Asn Cys

260 265 270

Asn Tyr Asn Leu Thr Lys Leu Leu Ser Leu Phe Ser Val Asn Asp Phe

275 280 285

Thr Cys Ser Gln Ile Ser Pro Ala Ala Ile Ala Ser Asn Cys Tyr Ser

290 295 300

Ser Leu Ile Leu Asp Tyr Phe Ser Tyr Pro Leu Ser Met Lys Ser Asp

305 310 315 320

Leu Ser Val Ser Ser Ala Gly Pro Ile Ser Gln Phe Asn Tyr Lys Gln

325 330 335

Ser Phe Ser Asn Pro Thr Cys Leu Ile Leu Ala Thr Val Pro His Asn

340 345 350

Leu Thr Thr Ile Thr Lys Pro Leu Lys Tyr Ser Tyr Ile Asn Lys Cys

355 360 365

Ser Arg Leu Leu Ser Asp Asp Arg Thr Glu Val Pro Gln Leu Val Asn

370 375 380

Ala Asn Gln Tyr Ser Pro Cys Val Ser Ile Val Pro Ser Thr Val Trp

385 390 395 400

Glu Asp Gly Asp Tyr Tyr Arg Lys Gln Leu Ser Pro Leu Glu Gly Gly

405 410 415

Gly Trp Leu Val Ala Ser Gly Ser Thr Val Ala Met Thr Glu Gln Leu

420 425 430

Gln Met Gly Phe Gly Ile Thr Val Gln Tyr Gly Thr Asp Thr Asn Ser

435 440 445

Val Cys Pro Lys Leu Glu Phe Ala Asn Asp Thr Lys Ile Ala Ser Gln

450 455 460

Leu Gly Asn Cys Val Glu Tyr Ser Gly Gly Gly Ser Gly Gly Gly Ser

465 470 475 480

Gly Gly Gly Ser Gly Gly Gly Ser Arg Phe Pro Asn Ile Thr Asn Leu

485 490 495

Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Lys Phe Pro Ser Val Tyr

500 505 510

Ala Trp Glu Arg Lys Lys Ile Ser Asn Cys Val Ala Asp Tyr Ser Val

515 520 525

Leu Tyr Asn Ser Thr Phe Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser

530 535 540

Ala Thr Lys Leu Asn Asp Leu Cys Phe Ser Asn Val Tyr Ala Asp Ser

545 550 555 560

Phe Val Val Lys Gly Asp Asp Val Arg Gln Ile Ala Pro Gly Gln Thr

565 570 575

Gly Val Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Met Gly

580 585 590

Cys Val Leu Ala Trp Asn Thr Arg Asn Ile Asp Ala Thr Ser Thr Gly

595 600 605

Asn Tyr Asn Tyr Lys Tyr Arg Tyr Leu Arg His Gly Lys Leu Arg Pro

610 615 620

Phe Glu Arg Asp Ile Ser Asn Val Pro Phe Ser Pro Asp Gly Lys Pro

625 630 635 640

Cys Thr Pro Pro Ala Leu Asn Cys Tyr Trp Pro Leu Asn Asp Tyr Gly

645 650 655

Phe Tyr Thr Thr Thr Gly Ile Gly Tyr Gln Pro Tyr Arg Val Val Val

660 665 670

Leu Ser Phe Glu Leu Leu Asn Ala Pro Ala Thr Val Cys Gly Pro Lys

675 680 685

Leu Ser Ala Ile Gly Gly Tyr Ile Pro Glu Ala Pro Arg Asp Gly Gln

690 695 700

Ala Tyr Val Arg Lys Asp Gly Glu Trp Val Leu Leu Ser Thr Phe Leu

705 710 715 720

His His His His His His

725

<210> 81

<211> 720

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 81

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Arg Phe Pro

1 5 10 15

Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg

20 25 30

Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val

35 40 45

Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys

50 55 60

Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn

65 70 75 80

Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile

85 90 95

Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro

100 105 110

Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp

115 120 125

Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys

130 135 140

Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln

145 150 155 160

Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe

165 170 175

Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln

180 185 190

Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala

195 200 205

Thr Val Cys Gly Pro Lys Lys Ser Gly Gly Gly Ser Gly Gly Gly Ser

210 215 220

Gly Gly Gly Ser Gly Gly Gly Ser Phe Glu Ala Lys Pro Ser Gly Ser

225 230 235 240

Val Val Ala Glu Gly Val Glu Cys Asp Phe Ser Pro Leu Leu Ser Gly

245 250 255

Thr Pro Pro Gln Val Tyr Asn Phe Lys Arg Leu Val Phe Thr Asn Cys

260 265 270

Asn Tyr Asn Leu Thr Lys Leu Leu Ser Leu Phe Ser Val Asn Asp Phe

275 280 285

Thr Cys Ser Gln Ile Ser Pro Ala Ala Ile Ala Ser Asn Cys Tyr Ser

290 295 300

Ser Leu Ile Leu Asp Tyr Phe Ser Tyr Pro Leu Ser Met Lys Ser Asp

305 310 315 320

Leu Ser Val Ser Ser Ala Gly Pro Ile Ser Gln Phe Asn Tyr Lys Gln

325 330 335

Ser Phe Ser Asn Pro Thr Cys Leu Ile Leu Ala Thr Val Pro His Asn

340 345 350

Leu Thr Thr Ile Thr Lys Pro Leu Lys Tyr Ser Tyr Ile Asn Lys Cys

355 360 365

Ser Arg Leu Leu Ser Asp Asp Arg Thr Glu Val Pro Gln Leu Val Asn

370 375 380

Ala Asn Gln Tyr Ser Pro Cys Val Ser Ile Val Pro Ser Thr Val Trp

385 390 395 400

Glu Asp Gly Asp Tyr Tyr Arg Lys Gln Leu Ser Pro Leu Glu Gly Gly

405 410 415

Gly Trp Leu Val Ala Ser Gly Ser Thr Val Ala Met Thr Glu Gln Leu

420 425 430

Gln Met Gly Phe Gly Ile Thr Val Gln Tyr Gly Thr Asp Thr Asn Ser

435 440 445

Val Cys Pro Lys Leu Glu Phe Ala Asn Asp Thr Lys Ile Ala Ser Gln

450 455 460

Leu Gly Asn Cys Val Glu Tyr Ser Gly Gly Gly Ser Gly Gly Gly Ser

465 470 475 480

Gly Gly Gly Ser Gly Gly Gly Ser Arg Phe Pro Asn Ile Thr Asn Leu

485 490 495

Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Lys Phe Pro Ser Val Tyr

500 505 510

Ala Trp Glu Arg Lys Lys Ile Ser Asn Cys Val Ala Asp Tyr Ser Val

515 520 525

Leu Tyr Asn Ser Thr Phe Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser

530 535 540

Ala Thr Lys Leu Asn Asp Leu Cys Phe Ser Asn Val Tyr Ala Asp Ser

545 550 555 560

Phe Val Val Lys Gly Asp Asp Val Arg Gln Ile Ala Pro Gly Gln Thr

565 570 575

Gly Val Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Met Gly

580 585 590

Cys Val Leu Ala Trp Asn Thr Arg Asn Ile Asp Ala Thr Ser Thr Gly

595 600 605

Asn Tyr Asn Tyr Lys Tyr Arg Tyr Leu Arg His Gly Lys Leu Arg Pro

610 615 620

Phe Glu Arg Asp Ile Ser Asn Val Pro Phe Ser Pro Asp Gly Lys Pro

625 630 635 640

Cys Thr Pro Pro Ala Leu Asn Cys Tyr Trp Pro Leu Asn Asp Tyr Gly

645 650 655

Phe Tyr Thr Thr Thr Gly Ile Gly Tyr Gln Pro Tyr Arg Val Val Val

660 665 670

Leu Ser Phe Glu Leu Leu Asn Ala Pro Ala Thr Val Cys Gly Pro Lys

675 680 685

Leu Ser Ala Ile Gly Gly Tyr Ile Pro Glu Ala Pro Arg Asp Gly Gln

690 695 700

Ala Tyr Val Arg Lys Asp Gly Glu Trp Val Leu Leu Ser Thr Phe Leu

705 710 715 720

<210> 82

<211> 713

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 82

Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn

1 5 10 15

Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser

20 25 30

Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser

35 40 45

Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys

50 55 60

Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val

65 70 75 80

Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr

85 90 95

Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn

100 105 110

Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu

115 120 125

Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu

130 135 140

Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn

145 150 155 160

Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val

165 170 175

Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His

180 185 190

Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Gly Gly Gly Ser Gly

195 200 205

Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Phe Glu Ala Lys Pro

210 215 220

Ser Gly Ser Val Val Ala Glu Gly Val Glu Cys Asp Phe Ser Pro Leu

225 230 235 240

Leu Ser Gly Thr Pro Pro Gln Val Tyr Asn Phe Lys Arg Leu Val Phe

245 250 255

Thr Asn Cys Asn Tyr Asn Leu Thr Lys Leu Leu Ser Leu Phe Ser Val

260 265 270

Asn Asp Phe Thr Cys Ser Gln Ile Ser Pro Ala Ala Ile Ala Ser Asn

275 280 285

Cys Tyr Ser Ser Leu Ile Leu Asp Tyr Phe Ser Tyr Pro Leu Ser Met

290 295 300

Lys Ser Asp Leu Ser Val Ser Ser Ala Gly Pro Ile Ser Gln Phe Asn

305 310 315 320

Tyr Lys Gln Ser Phe Ser Asn Pro Thr Cys Leu Ile Leu Ala Thr Val

325 330 335

Pro His Asn Leu Thr Thr Ile Thr Lys Pro Leu Lys Tyr Ser Tyr Ile

340 345 350

Asn Lys Cys Ser Arg Leu Leu Ser Asp Asp Arg Thr Glu Val Pro Gln

355 360 365

Leu Val Asn Ala Asn Gln Tyr Ser Pro Cys Val Ser Ile Val Pro Ser

370 375 380

Thr Val Trp Glu Asp Gly Asp Tyr Tyr Arg Lys Gln Leu Ser Pro Leu

385 390 395 400

Glu Gly Gly Gly Trp Leu Val Ala Ser Gly Ser Thr Val Ala Met Thr

405 410 415

Glu Gln Leu Gln Met Gly Phe Gly Ile Thr Val Gln Tyr Gly Thr Asp

420 425 430

Thr Asn Ser Val Cys Pro Lys Leu Glu Phe Ala Asn Asp Thr Lys Ile

435 440 445

Ala Ser Gln Leu Gly Asn Cys Val Glu Tyr Ser Gly Gly Gly Ser Gly

450 455 460

Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Phe Pro Asn Ile

465 470 475 480

Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Lys Phe Pro

485 490 495

Ser Val Tyr Ala Trp Glu Arg Lys Lys Ile Ser Asn Cys Val Ala Asp

500 505 510

Tyr Ser Val Leu Tyr Asn Ser Thr Phe Phe Ser Thr Phe Lys Cys Tyr

515 520 525

Gly Val Ser Ala Thr Lys Leu Asn Asp Leu Cys Phe Ser Asn Val Tyr

530 535 540

Ala Asp Ser Phe Val Val Lys Gly Asp Asp Val Arg Gln Ile Ala Pro

545 550 555 560

Gly Gln Thr Gly Val Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp

565 570 575

Phe Met Gly Cys Val Leu Ala Trp Asn Thr Arg Asn Ile Asp Ala Thr

580 585 590

Ser Thr Gly Asn Tyr Asn Tyr Lys Tyr Arg Tyr Leu Arg His Gly Lys

595 600 605

Leu Arg Pro Phe Glu Arg Asp Ile Ser Asn Val Pro Phe Ser Pro Asp

610 615 620

Gly Lys Pro Cys Thr Pro Pro Ala Leu Asn Cys Tyr Trp Pro Leu Asn

625 630 635 640

Asp Tyr Gly Phe Tyr Thr Thr Thr Gly Ile Gly Tyr Gln Pro Tyr Arg

645 650 655

Val Val Val Leu Ser Phe Glu Leu Leu Asn Ala Pro Ala Thr Val Cys

660 665 670

Gly Pro Lys Leu Ser Ala Ile Gly Gly Tyr Ile Pro Glu Ala Pro Arg

675 680 685

Asp Gly Gln Ala Tyr Val Arg Lys Asp Gly Glu Trp Val Leu Leu Ser

690 695 700

Thr Phe Leu His His His His His His

705 710

<210> 83

<211> 707

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 83

Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn

1 5 10 15

Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser

20 25 30

Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser

35 40 45

Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys

50 55 60

Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val

65 70 75 80

Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr

85 90 95

Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn

100 105 110

Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu

115 120 125

Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu

130 135 140

Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn

145 150 155 160

Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val

165 170 175

Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His

180 185 190

Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Gly Gly Gly Ser Gly

195 200 205

Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Phe Glu Ala Lys Pro

210 215 220

Ser Gly Ser Val Val Ala Glu Gly Val Glu Cys Asp Phe Ser Pro Leu

225 230 235 240

Leu Ser Gly Thr Pro Pro Gln Val Tyr Asn Phe Lys Arg Leu Val Phe

245 250 255

Thr Asn Cys Asn Tyr Asn Leu Thr Lys Leu Leu Ser Leu Phe Ser Val

260 265 270

Asn Asp Phe Thr Cys Ser Gln Ile Ser Pro Ala Ala Ile Ala Ser Asn

275 280 285

Cys Tyr Ser Ser Leu Ile Leu Asp Tyr Phe Ser Tyr Pro Leu Ser Met

290 295 300

Lys Ser Asp Leu Ser Val Ser Ser Ala Gly Pro Ile Ser Gln Phe Asn

305 310 315 320

Tyr Lys Gln Ser Phe Ser Asn Pro Thr Cys Leu Ile Leu Ala Thr Val

325 330 335

Pro His Asn Leu Thr Thr Ile Thr Lys Pro Leu Lys Tyr Ser Tyr Ile

340 345 350

Asn Lys Cys Ser Arg Leu Leu Ser Asp Asp Arg Thr Glu Val Pro Gln

355 360 365

Leu Val Asn Ala Asn Gln Tyr Ser Pro Cys Val Ser Ile Val Pro Ser

370 375 380

Thr Val Trp Glu Asp Gly Asp Tyr Tyr Arg Lys Gln Leu Ser Pro Leu

385 390 395 400

Glu Gly Gly Gly Trp Leu Val Ala Ser Gly Ser Thr Val Ala Met Thr

405 410 415

Glu Gln Leu Gln Met Gly Phe Gly Ile Thr Val Gln Tyr Gly Thr Asp

420 425 430

Thr Asn Ser Val Cys Pro Lys Leu Glu Phe Ala Asn Asp Thr Lys Ile

435 440 445

Ala Ser Gln Leu Gly Asn Cys Val Glu Tyr Ser Gly Gly Gly Ser Gly

450 455 460

Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Phe Pro Asn Ile

465 470 475 480

Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Lys Phe Pro

485 490 495

Ser Val Tyr Ala Trp Glu Arg Lys Lys Ile Ser Asn Cys Val Ala Asp

500 505 510

Tyr Ser Val Leu Tyr Asn Ser Thr Phe Phe Ser Thr Phe Lys Cys Tyr

515 520 525

Gly Val Ser Ala Thr Lys Leu Asn Asp Leu Cys Phe Ser Asn Val Tyr

530 535 540

Ala Asp Ser Phe Val Val Lys Gly Asp Asp Val Arg Gln Ile Ala Pro

545 550 555 560

Gly Gln Thr Gly Val Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp

565 570 575

Phe Met Gly Cys Val Leu Ala Trp Asn Thr Arg Asn Ile Asp Ala Thr

580 585 590

Ser Thr Gly Asn Tyr Asn Tyr Lys Tyr Arg Tyr Leu Arg His Gly Lys

595 600 605

Leu Arg Pro Phe Glu Arg Asp Ile Ser Asn Val Pro Phe Ser Pro Asp

610 615 620

Gly Lys Pro Cys Thr Pro Pro Ala Leu Asn Cys Tyr Trp Pro Leu Asn

625 630 635 640

Asp Tyr Gly Phe Tyr Thr Thr Thr Gly Ile Gly Tyr Gln Pro Tyr Arg

645 650 655

Val Val Val Leu Ser Phe Glu Leu Leu Asn Ala Pro Ala Thr Val Cys

660 665 670

Gly Pro Lys Leu Ser Ala Ile Gly Gly Tyr Ile Pro Glu Ala Pro Arg

675 680 685

Asp Gly Gln Ala Tyr Val Arg Lys Asp Gly Glu Trp Val Leu Leu Ser

690 695 700

Thr Phe Leu

705

<210> 84

<211> 1273

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 84

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg Ser Val Ala

675 680 685

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser

690 695 700

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

705 710 715 720

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

725 730 735

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

740 745 750

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

755 760 765

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

770 775 780

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe

785 790 795 800

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

805 810 815

Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly

820 825 830

Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp

835 840 845

Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu

850 855 860

Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly

865 870 875 880

Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile

885 890 895

Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr

900 905 910

Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn

915 920 925

Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala

930 935 940

Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn

945 950 955 960

Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val

965 970 975

Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln

980 985 990

Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val

995 1000 1005

Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn

1010 1015 1020

Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

1025 1030 1035

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro

1040 1045 1050

Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val

1055 1060 1065

Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

1070 1075 1080

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

1085 1090 1095

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln

1100 1105 1110

Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val

1115 1120 1125

Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro

1130 1135 1140

Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn

1145 1150 1155

His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn

1160 1165 1170

Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu

1175 1180 1185

Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu

1190 1195 1200

Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu

1205 1210 1215

Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met

1220 1225 1230

Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys

1235 1240 1245

Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro

1250 1255 1260

Val Leu Lys Gly Val Lys Leu His Tyr Thr

1265 1270

<210> 85

<211> 1260

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 85

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg

660 665 670

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala

675 680 685

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn

690 695 700

Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys

705 710 715 720

Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys

725 730 735

Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg

740 745 750

Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val

755 760 765

Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe

770 775 780

Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser

785 790 795 800

Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala

805 810 815

Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala

820 825 830

Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu

835 840 845

Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu

850 855 860

Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala

865 870 875 880

Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile

885 890 895

Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn

900 905 910

Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr

915 920 925

Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln

930 935 940

Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile

945 950 955 960

Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala

965 970 975

Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln

980 985 990

Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser

995 1000 1005

Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln

1010 1015 1020

Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser

1025 1030 1035

Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr

1040 1045 1050

Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile

1055 1060 1065

Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val

1070 1075 1080

Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu

1085 1090 1095

Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys

1100 1105 1110

Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu

1115 1120 1125

Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe

1130 1135 1140

Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly

1145 1150 1155

Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu

1160 1165 1170

Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln

1175 1180 1185

Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile

1190 1195 1200

Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr

1205 1210 1215

Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly

1220 1225 1230

Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser

1235 1240 1245

Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255 1260

<210> 86

<211> 1273

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 86

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln Ser Val Ala

675 680 685

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser

690 695 700

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

705 710 715 720

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

725 730 735

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

740 745 750

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

755 760 765

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

770 775 780

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe

785 790 795 800

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

805 810 815

Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly

820 825 830

Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp

835 840 845

Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu

850 855 860

Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly

865 870 875 880

Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile

885 890 895

Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr

900 905 910

Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn

915 920 925

Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala

930 935 940

Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn

945 950 955 960

Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val

965 970 975

Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln

980 985 990

Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val

995 1000 1005

Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn

1010 1015 1020

Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

1025 1030 1035

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro

1040 1045 1050

Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val

1055 1060 1065

Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

1070 1075 1080

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

1085 1090 1095

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln

1100 1105 1110

Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val

1115 1120 1125

Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro

1130 1135 1140

Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn

1145 1150 1155

His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn

1160 1165 1170

Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu

1175 1180 1185

Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu

1190 1195 1200

Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu

1205 1210 1215

Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met

1220 1225 1230

Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys

1235 1240 1245

Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro

1250 1255 1260

Val Leu Lys Gly Val Lys Leu His Tyr Thr

1265 1270

<210> 87

<211> 1260

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 87

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln

660 665 670

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala

675 680 685

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn

690 695 700

Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys

705 710 715 720

Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys

725 730 735

Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg

740 745 750

Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val

755 760 765

Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe

770 775 780

Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser

785 790 795 800

Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala

805 810 815

Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala

820 825 830

Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu

835 840 845

Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu

850 855 860

Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala

865 870 875 880

Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile

885 890 895

Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn

900 905 910

Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr

915 920 925

Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln

930 935 940

Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile

945 950 955 960

Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala

965 970 975

Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln

980 985 990

Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser

995 1000 1005

Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln

1010 1015 1020

Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser

1025 1030 1035

Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr

1040 1045 1050

Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile

1055 1060 1065

Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val

1070 1075 1080

Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu

1085 1090 1095

Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys

1100 1105 1110

Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu

1115 1120 1125

Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe

1130 1135 1140

Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly

1145 1150 1155

Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu

1160 1165 1170

Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln

1175 1180 1185

Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile

1190 1195 1200

Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr

1205 1210 1215

Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly

1220 1225 1230

Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser

1235 1240 1245

Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255 1260

<210> 88

<211> 1219

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 88

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln Ser Val Ala

675 680 685

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser

690 695 700

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

705 710 715 720

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

725 730 735

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

740 745 750

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

755 760 765

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

770 775 780

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe

785 790 795 800

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

805 810 815

Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly

820 825 830

Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp

835 840 845

Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu

850 855 860

Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly

865 870 875 880

Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile

885 890 895

Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr

900 905 910

Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn

915 920 925

Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala

930 935 940

Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn

945 950 955 960

Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val

965 970 975

Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln

980 985 990

Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val

995 1000 1005

Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn

1010 1015 1020

Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

1025 1030 1035

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro

1040 1045 1050

Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val

1055 1060 1065

Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

1070 1075 1080

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

1085 1090 1095

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln

1100 1105 1110

Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val

1115 1120 1125

Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro

1130 1135 1140

Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn

1145 1150 1155

His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn

1160 1165 1170

Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu

1175 1180 1185

Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu

1190 1195 1200

Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro His His His His His

1205 1210 1215

His

<210> 89

<211> 1206

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 89

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln

660 665 670

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala

675 680 685

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn

690 695 700

Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys

705 710 715 720

Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys

725 730 735

Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg

740 745 750

Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val

755 760 765

Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe

770 775 780

Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser

785 790 795 800

Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala

805 810 815

Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala

820 825 830

Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu

835 840 845

Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu

850 855 860

Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala

865 870 875 880

Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile

885 890 895

Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn

900 905 910

Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr

915 920 925

Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln

930 935 940

Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile

945 950 955 960

Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala

965 970 975

Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln

980 985 990

Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser

995 1000 1005

Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln

1010 1015 1020

Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser

1025 1030 1035

Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr

1040 1045 1050

Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile

1055 1060 1065

Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val

1070 1075 1080

Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu

1085 1090 1095

Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys

1100 1105 1110

Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu

1115 1120 1125

Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe

1130 1135 1140

Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly

1145 1150 1155

Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu

1160 1165 1170

Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln

1175 1180 1185

Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro His His His

1190 1195 1200

His His His

1205

<210> 90

<211> 17

<212> PRT

<213> 人工序列

<220>

<223> 接头

<400> 90

Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly

1 5 10 15

Ser

<210> 91

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 接头

<400> 91

Ser Gly Gly Gly

1

<210> 92

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 接头

<400> 92

Gly Gly Gly Ser

1

<210> 93

<211> 5

<212> PRT

<213> 人工序列

<220>

<223> 接头

<400> 93

Gly Gly Gly Gly Ser

1 5

<210> 94

<211> 90

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 94

Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn Gly

1 5 10 15

Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro

20 25 30

Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val Arg

35 40 45

Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly

50 55 60

Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val Ala

65 70 75 80

Val Leu Tyr Gln Asp Val Asn Cys Thr Glu

85 90

<210> 95

<211> 3639

<212> DNA

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 95

atgttcgtct tcctggtgct gctgcccctg gtgtccagcc agtgcgtgaa cctgaccact 60

aggactcagc tgcctcccgc ttacaccaac tcattcactc gcggtgtgta ctaccctgac 120

aaggtcttcc gttcttcagt gctgcactca actcaggacc tgttcctgcc cttcttctcc 180

aacgtcacct ggttccacgc catccacgtg tccggcacca acggcactaa gcgcttcgac 240

aacccagtgc tgcctttcaa cgacggtgtc tacttcgctt caaccgagaa gtccaacatc 300

atccgtggat ggatcttcgg caccactctg gacagcaaga ctcagtctct gctgatcgtc 360

aacaacgcca ccaacgtggt catcaaggtc tgcgaattcc agttctgcaa cgacccattc 420

ctgggcgtct actaccacaa gaacaacaag tcatggatgg agtccgaatt ccgcgtctac 480

tccagcgcta acaactgcac tttcgagtac gtgtcccagc ctttcctgat ggacctggaa 540

ggaaagcagg gtaacttcaa gaacctgagg gagttcgtgt tcaagaacat cgacggatac 600

ttcaagattt acagcaagca caccccaatc aacctggtgc gcgacctgcc tcagggtttc 660

tctgctctgg agccactggt ggacctgcct atcggcatca acatcacccg cttccagact 720

ctgctggctc tgcaccgttc ctacctgact ccaggcgact catcttctgg atggactgct 780

ggagctgctg cttactacgt gggctacctg cagcctcgca ccttcctgct gaagtacaac 840

gaaaacggaa ccatcactga cgccgtcgac tgcgctctgg accctctgtc agaaaccaag 900

tgcactctga agtccttcac cgtggagaag ggcatctacc agacttcaaa cttcagggtg 960

cagcccaccg aatccatcgt cagattccct aacatcacta acctgtgccc cttcggagag 1020

gtcttcaacg ccacccgctt cgcttccgtg tacgcctgga acaggaagag aatctcaaac 1080

tgcgtcgctg actactccgt gctgtacaac tcagcctcct tcagcacctt caagtgctac 1140

ggcgtgtcac caactaagct gaacgacctg tgcttcacca acgtctacgc cgactccttc 1200

gtgatcaggg gagacgaggt cagacagatc gctcctggcc agactggaaa gatcgccgac 1260

tacaactaca agctgcccga cgacttcacc ggttgcgtca tcgcttggaa cagcaacaac 1320

ctggactcta aagtgggtgg caactacaac tacctgtacc gcctgttccg taagtcaaac 1380

ctgaagccat tcgagaggga catcagcact gaaatctacc aggccggatc taccccttgc 1440

aacggtgtcg agggcttcaa ctgctacttc cccctgcagt cctacggttt ccagccaact 1500

aacggtgtgg gctaccagcc ttacagagtg gtcgtgctga gcttcgaact gctccacgct 1560

cctgctactg tgtgcggtcc aaagaagtct accaacctgg tcaagaacaa gtgcgtgaac 1620

ttcaacttca acggcctgac cggaactggt gtcctgaccg agagcaacaa gaagttcctg 1680

cccttccagc agttcggaag ggacatcgct gacaccactg acgctgtgcg cgaccctcag 1740

accctggaaa tcctggacat cactccatgc tcattcggag gtgtctccgt gatcacccct 1800

ggcaccaaca cttctaacca ggtcgctgtg ctgtaccagg acgtcaactg caccgaggtc 1860

cctgtggcca tccacgctga ccagctgacc cccacttggc gcgtgtactc caccggcagc 1920

aacgtgttcc agactcgtgc tggttgcctg atcggcgccg agcacgtgaa caacagctac 1980

gaatgcgaca tccccatcgg cgctggaatc tgcgcctctt accagaccca gactaacagc 2040

ccacgcaggg ctcgctctgt ggcctctcag tcaatcatcg cttacaccat gtcactgggc 2100

gctgaaaact ccgtggccta ctctaacaac tcaatcgcca tccccaccaa cttcactatc 2160

agcgtgacca ctgagatcct gccagtcagc atgaccaaga cttctgtgga ctgcactatg 2220

tacatctgcg gagacagcac cgaatgctct aacctgctgc tgcagtacgg ctctttctgc 2280

acccagctga accgtgctct gactggaatc gccgtggagc aggacaagaa cactcaggaa 2340

gtcttcgctc aggtgaagca aatctacaag accccaccta tcaaggactt cggcggattc 2400

aacttctccc agatcctgcc tgacccctcc aagccaagca agcgctcttt catcgaggac 2460

ctgctgttca acaaggtcac tctggccgac gctggattca tcaagcagta cggagactgc 2520

ctgggtgaca tcgccgctcg tgacctgatc tgcgctcaga agttcaacgg tctgaccgtg 2580

ctgcccccac tgctgactga cgaaatgatc gcccagtaca ctagcgccct gctggctgga 2640

accatcactt ctggttggac cttcggtgct ggcgccgctc tgcagatccc tttcgctatg 2700

cagatggcct accgtttcaa cggaatcggt gtcacccaga acgtgctgta cgagaaccag 2760

aagctgatcg ctaaccagtt caactcagcc atcggaaaga tccaggacag cctgagctct 2820

actgcctctg ctctgggcaa gctgcaggac gtcgtgaacc agaacgccca ggctctgaac 2880

accctggtca agcagctgtc atccaacttc ggtgctatca gctctgtgct gaacgacatc 2940

ctgtcccgcc tggacaaggt cgaggccgaa gtgcagatcg accgcctgat cactggccgt 3000

ctgcagtcac tgcagaccta cgtgactcag cagctgatca gggccgctga aatcagagcc 3060

tccgctaacc tggccgctac caagatgagc gagtgcgtcc tgggtcaatc taagcgtgtg 3120

gacttctgcg gcaagggata ccacctgatg tcattccctc agtctgctcc ccacggtgtg 3180

gtgttcctgc acgtcaccta cgtgccagcc caggagaaga acttcaccac tgcccctgct 3240

atctgccacg acggcaaggc tcacttcccc agggaaggtg tcttcgtgag caacggcacc 3300

cactggttcg tcactcagag aaacttctac gagccacaga tcatcaccac tgacaacact 3360

ttcgtgtctg gaaactgcga cgtggtcatc ggtatcgtca acaacaccgt gtacgacccc 3420

ctgcagccag agctggactc attcaaggag gaactggaca agtacttcaa gaaccacacc 3480

tcccctgacg tcgacctggg cgacatctca ggaatcaacg cttccgtcgt gaacatccag 3540

aaggagatcg accgcctgaa cgaagtggcc aagaacctga acgaaagcct gatcgacctg 3600

caggagctgg gcaagtacga acagtacatc aagtggcct 3639

<210> 96

<211> 3822

<212> DNA

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 96

atgttcgtct tcctggtgct gctgcccctg gtgtccagcc agtgcgtgaa cctgaccact 60

aggactcagc tgcctcccgc ttacaccaac tcattcactc gcggtgtgta ctaccctgac 120

aaggtcttcc gttcttcagt gctgcactca actcaggacc tgttcctgcc cttcttctcc 180

aacgtcacct ggttccacgc catccacgtg tccggcacca acggcactaa gcgcttcgac 240

aacccagtgc tgcctttcaa cgacggtgtc tacttcgctt caaccgagaa gtccaacatc 300

atccgtggat ggatcttcgg caccactctg gacagcaaga ctcagtctct gctgatcgtc 360

aacaacgcca ccaacgtggt catcaaggtc tgcgaattcc agttctgcaa cgacccattc 420

ctgggcgtct actaccacaa gaacaacaag tcatggatgg agtccgaatt ccgcgtctac 480

tccagcgcta acaactgcac tttcgagtac gtgtcccagc ctttcctgat ggacctggaa 540

ggaaagcagg gtaacttcaa gaacctgagg gagttcgtgt tcaagaacat cgacggatac 600

ttcaagattt acagcaagca caccccaatc aacctggtgc gcgacctgcc tcagggtttc 660

tctgctctgg agccactggt ggacctgcct atcggcatca acatcacccg cttccagact 720

ctgctggctc tgcaccgttc ctacctgact ccaggcgact catcttctgg atggactgct 780

ggagctgctg cttactacgt gggctacctg cagcctcgca ccttcctgct gaagtacaac 840

gaaaacggaa ccatcactga cgccgtcgac tgcgctctgg accctctgtc agaaaccaag 900

tgcactctga agtccttcac cgtggagaag ggcatctacc agacttcaaa cttcagggtg 960

cagcccaccg aatccatcgt cagattccct aacatcacta acctgtgccc cttcggagag 1020

gtcttcaacg ccacccgctt cgcttccgtg tacgcctgga acaggaagag aatctcaaac 1080

tgcgtcgctg actactccgt gctgtacaac tcagcctcct tcagcacctt caagtgctac 1140

ggcgtgtcac caactaagct gaacgacctg tgcttcacca acgtctacgc cgactccttc 1200

gtgatcaggg gagacgaggt cagacagatc gctcctggcc agactggaaa gatcgccgac 1260

tacaactaca agctgcccga cgacttcacc ggttgcgtca tcgcttggaa cagcaacaac 1320

ctggactcta aagtgggtgg caactacaac tacctgtacc gcctgttccg taagtcaaac 1380

ctgaagccat tcgagaggga catcagcact gaaatctacc aggccggatc taccccttgc 1440

aacggtgtcg agggcttcaa ctgctacttc cccctgcagt cctacggttt ccagccaact 1500

aacggtgtgg gctaccagcc ttacagagtg gtcgtgctga gcttcgaact gctccacgct 1560

cctgctactg tgtgcggtcc aaagaagtct accaacctgg tcaagaacaa gtgcgtgaac 1620

ttcaacttca acggcctgac cggaactggt gtcctgaccg agagcaacaa gaagttcctg 1680

cccttccagc agttcggaag ggacatcgct gacaccactg acgctgtgcg cgaccctcag 1740

accctggaaa tcctggacat cactccatgc tcattcggag gtgtctccgt gatcacccct 1800

ggcaccaaca cttctaacca ggtcgctgtg ctgtaccagg acgtcaactg caccgaggtc 1860

cctgtggcca tccacgctga ccagctgacc cccacttggc gcgtgtactc caccggcagc 1920

aacgtgttcc agactcgtgc tggttgcctg atcggcgccg agcacgtgaa caacagctac 1980

gaatgcgaca tccccatcgg cgctggaatc tgcgcctctt accagaccca gactaacagc 2040

ccacagcagg ctcagtctgt ggcctctcag tcaatcatcg cttacaccat gtcactgggc 2100

gctgaaaact ccgtggccta ctctaacaac tcaatcgcca tccccaccaa cttcactatc 2160

agcgtgacca ctgagatcct gccagtcagc atgaccaaga cttctgtgga ctgcactatg 2220

tacatctgcg gagacagcac cgaatgctct aacctgctgc tgcagtacgg ctctttctgc 2280

acccagctga accgtgctct gactggaatc gccgtggagc aggacaagaa cactcaggaa 2340

gtcttcgctc aggtgaagca aatctacaag accccaccta tcaaggactt cggcggattc 2400

aacttctccc agatcctgcc tgacccctcc aagccaagca agcgctcttt catcgaggac 2460

ctgctgttca acaaggtcac tctggccgac gctggattca tcaagcagta cggagactgc 2520

ctgggtgaca tcgccgctcg tgacctgatc tgcgctcaga agttcaacgg tctgaccgtg 2580

ctgcccccac tgctgactga cgaaatgatc gcccagtaca ctagcgccct gctggctgga 2640

accatcactt ctggttggac cttcggtgct ggcgccgctc tgcagatccc tttcgctatg 2700

cagatggcct accgtttcaa cggaatcggt gtcacccaga acgtgctgta cgagaaccag 2760

aagctgatcg ctaaccagtt caactcagcc atcggaaaga tccaggacag cctgagctct 2820

actgcctctg ctctgggcaa gctgcaggac gtcgtgaacc agaacgccca ggctctgaac 2880

accctggtca agcagctgtc atccaacttc ggtgctatca gctctgtgct gaacgacatc 2940

ctgtcccgcc tggaccctcc cgaggccgaa gtgcagatcg accgcctgat cactggccgt 3000

ctgcagtcac tgcagaccta cgtgactcag cagctgatca gggccgctga aatcagagcc 3060

tccgctaacc tggccgctac caagatgagc gagtgcgtcc tgggtcaatc taagcgtgtg 3120

gacttctgcg gcaagggata ccacctgatg tcattccctc agtctgctcc ccacggtgtg 3180

gtgttcctgc acgtcaccta cgtgccagcc caggagaaga acttcaccac tgcccctgct 3240

atctgccacg acggcaaggc tcacttcccc agggaaggtg tcttcgtgag caacggcacc 3300

cactggttcg tcactcagag aaacttctac gagccacaga tcatcaccac tgacaacact 3360

ttcgtgtctg gaaactgcga cgtggtcatc ggtatcgtca acaacaccgt gtacgacccc 3420

ctgcagccag agctggactc attcaaggag gaactggaca agtacttcaa gaaccacacc 3480

tcccctgacg tcgacctggg cgacatctca ggaatcaacg cttccgtcgt gaacatccag 3540

aaggagatcg accgcctgaa cgaagtggcc aagaacctga acgaaagcct gatcgacctg 3600

caggagctgg gcaagtacga acagtacatc aagtggcctt ggtacatctg gctgggtttc 3660

atcgctggcc tcatcgctat cgtgatggtg accatcatgc tgtgctgcat gacttcatgc 3720

tgctcctgcc tgaagggctg ctgcagctgc ggatcttgct gcaagttcga cgaggacgac 3780

tctgaacccg tcctgaaggg cgtgaagctg cactacacct aa 3822

<210> 97

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 97

Gly Ser Ala Ser

1

<210> 98

<211> 8

<212> PRT

<213> 人工序列

<220>

<223> HRV3C 切割位点

<400> 98

Leu Glu Val Leu Phe Gln Gly Pro

1 5

<210> 99

<211> 8

<212> PRT

<213> 人工序列

<220>

<223> Strep-标签

<400> 99

Trp Ser His Pro Gln Phe Glu Lys

1 5

<210> 100

<211> 6

<212> PRT

<213> 人工序列

<220>

<223> 6X-His标签

<400> 100

His His His His His His

1 5

<210> 101

<211> 10

<212> PRT

<213> 人工序列

<220>

<223> His-标签

<220>

<221> 位点

<222> (6)..(6)

<223> 可存在或不存在

<220>

<221> 位点

<222> (7)..(7)

<223> 可存在或不存在

<220>

<221> 位点

<222> (8)..(8)

<223> 可存在或不存在

<220>

<221> 位点

<222> (9)..(9)

<223> 可存在或不存在

<220>

<221> 位点

<222> (10)..(10)

<223> 可存在或不存在

<400> 101

His His His His His His His His His His

1 5 10

<210> 102

<211> 8

<212> PRT

<213> 人工序列

<220>

<223> 8X-His标签

<400> 102

His His His His His His His His

1 5

<210> 103

<211> 30

<212> PRT

<213> 人工序列

<220>

<223> 折叠子

<400> 103

Gly Tyr Ile Pro Glu Ala Pro Arg Asp Gly Gln Ala Tyr Val Arg Lys

1 5 10 15

Asp Gly Glu Trp Val Leu Leu Ser Thr Phe Leu Ser Pro Ala

20 25 30

<210> 104

<211> 10

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 104

Glu Asp Leu Leu Phe Asn Lys Val Thr Leu

1 5 10

<210> 105

<211> 1263

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 105

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln Ser Val Ala

675 680 685

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser

690 695 700

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

705 710 715 720

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

725 730 735

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

740 745 750

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

755 760 765

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

770 775 780

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe

785 790 795 800

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

805 810 815

Phe Ile Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly

820 825 830

Asp Ile Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu

835 840 845

Thr Val Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr

850 855 860

Ser Ala Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala

865 870 875 880

Gly Ala Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe

885 890 895

Asn Gly Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu

900 905 910

Ile Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu

915 920 925

Ser Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln

930 935 940

Asn Ala Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe

945 950 955 960

Gly Ala Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro

965 970 975

Pro Glu Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln

980 985 990

Ser Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile

995 1000 1005

Arg Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val

1010 1015 1020

Leu Gly Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His

1025 1030 1035

Leu Met Ser Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu

1040 1045 1050

His Val Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala

1055 1060 1065

Pro Ala Ile Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly

1070 1075 1080

Val Phe Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn

1085 1090 1095

Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser

1100 1105 1110

Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr

1115 1120 1125

Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp

1130 1135 1140

Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp

1145 1150 1155

Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile

1160 1165 1170

Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile

1175 1180 1185

Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro

1190 1195 1200

Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val

1205 1210 1215

Met Val Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys

1220 1225 1230

Leu Lys Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu

1235 1240 1245

Asp Asp Ser Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255 1260

<210> 106

<211> 1250

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 106

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln

660 665 670

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala

675 680 685

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn

690 695 700

Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys

705 710 715 720

Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys

725 730 735

Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg

740 745 750

Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val

755 760 765

Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe

770 775 780

Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser

785 790 795 800

Lys Arg Ser Phe Ile Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp

805 810 815

Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe

820 825 830

Asn Gly Leu Thr Val Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala

835 840 845

Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr

850 855 860

Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala

865 870 875 880

Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn

885 890 895

Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln

900 905 910

Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val

915 920 925

Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser

930 935 940

Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg

945 950 955 960

Leu Asp Pro Pro Glu Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly

965 970 975

Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala

980 985 990

Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu

995 1000 1005

Cys Val Leu Gly Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly

1010 1015 1020

Tyr His Leu Met Ser Phe Pro Gln Ser Ala Pro His Gly Val Val

1025 1030 1035

Phe Leu His Val Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr

1040 1045 1050

Thr Ala Pro Ala Ile Cys His Asp Gly Lys Ala His Phe Pro Arg

1055 1060 1065

Glu Gly Val Phe Val Ser Asn Gly Thr His Trp Phe Val Thr Gln

1070 1075 1080

Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe

1085 1090 1095

Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr

1100 1105 1110

Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu

1115 1120 1125

Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu

1130 1135 1140

Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys

1145 1150 1155

Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser

1160 1165 1170

Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys

1175 1180 1185

Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala

1190 1195 1200

Ile Val Met Val Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys

1205 1210 1215

Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe

1220 1225 1230

Asp Glu Asp Asp Ser Glu Pro Val Leu Lys Gly Val Lys Leu His

1235 1240 1245

Tyr Thr

1250

<210> 107

<211> 1263

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 107

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg Ser Val Ala

675 680 685

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser

690 695 700

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

705 710 715 720

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

725 730 735

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

740 745 750

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

755 760 765

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

770 775 780

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe

785 790 795 800

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

805 810 815

Phe Ile Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly

820 825 830

Asp Ile Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu

835 840 845

Thr Val Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr

850 855 860

Ser Ala Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala

865 870 875 880

Gly Ala Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe

885 890 895

Asn Gly Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu

900 905 910

Ile Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu

915 920 925

Ser Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln

930 935 940

Asn Ala Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe

945 950 955 960

Gly Ala Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys

965 970 975

Val Glu Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln

980 985 990

Ser Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile

995 1000 1005

Arg Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val

1010 1015 1020

Leu Gly Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His

1025 1030 1035

Leu Met Ser Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu

1040 1045 1050

His Val Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala

1055 1060 1065

Pro Ala Ile Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly

1070 1075 1080

Val Phe Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn

1085 1090 1095

Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser

1100 1105 1110

Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr

1115 1120 1125

Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp

1130 1135 1140

Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp

1145 1150 1155

Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile

1160 1165 1170

Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile

1175 1180 1185

Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro

1190 1195 1200

Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val

1205 1210 1215

Met Val Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys

1220 1225 1230

Leu Lys Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu

1235 1240 1245

Asp Asp Ser Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255 1260

<210> 108

<211> 1250

<212> PRT

<213> 人工序列

<220>

<223> 重组刺突蛋白

<400> 108

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg

660 665 670

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala

675 680 685

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn

690 695 700

Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys

705 710 715 720

Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys

725 730 735

Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg

740 745 750

Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val

755 760 765

Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe

770 775 780

Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser

785 790 795 800

Lys Arg Ser Phe Ile Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp

805 810 815

Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe

820 825 830

Asn Gly Leu Thr Val Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala

835 840 845

Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr

850 855 860

Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala

865 870 875 880

Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn

885 890 895

Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln

900 905 910

Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val

915 920 925

Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser

930 935 940

Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg

945 950 955 960

Leu Asp Lys Val Glu Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly

965 970 975

Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala

980 985 990

Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu

995 1000 1005

Cys Val Leu Gly Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly

1010 1015 1020

Tyr His Leu Met Ser Phe Pro Gln Ser Ala Pro His Gly Val Val

1025 1030 1035

Phe Leu His Val Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr

1040 1045 1050

Thr Ala Pro Ala Ile Cys His Asp Gly Lys Ala His Phe Pro Arg

1055 1060 1065

Glu Gly Val Phe Val Ser Asn Gly Thr His Trp Phe Val Thr Gln

1070 1075 1080

Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe

1085 1090 1095

Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr

1100 1105 1110

Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu

1115 1120 1125

Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu

1130 1135 1140

Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys

1145 1150 1155

Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser

1160 1165 1170

Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys

1175 1180 1185

Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala

1190 1195 1200

Ile Val Met Val Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys

1205 1210 1215

Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe

1220 1225 1230

Asp Glu Asp Asp Ser Glu Pro Val Leu Lys Gly Val Lys Leu His

1235 1240 1245

Tyr Thr

1250

<210> 109

<211> 1273

<212> PRT

<213> 人工序列

<220>

<223> 重组CoV S多肽

<400> 109

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gly Ser Ala Ser Ser Val Ala

675 680 685

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser

690 695 700

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

705 710 715 720

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

725 730 735

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

740 745 750

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

755 760 765

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

770 775 780

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe

785 790 795 800

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

805 810 815

Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly

820 825 830

Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp

835 840 845

Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu

850 855 860

Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly

865 870 875 880

Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile

885 890 895

Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr

900 905 910

Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn

915 920 925

Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala

930 935 940

Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn

945 950 955 960

Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val

965 970 975

Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln

980 985 990

Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val

995 1000 1005

Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn

1010 1015 1020

Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

1025 1030 1035

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro

1040 1045 1050

Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val

1055 1060 1065

Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

1070 1075 1080

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

1085 1090 1095

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln

1100 1105 1110

Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val

1115 1120 1125

Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro

1130 1135 1140

Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn

1145 1150 1155

His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn

1160 1165 1170

Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu

1175 1180 1185

Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu

1190 1195 1200

Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu

1205 1210 1215

Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met

1220 1225 1230

Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys

1235 1240 1245

Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro

1250 1255 1260

Val Leu Lys Gly Val Lys Leu His Tyr Thr

1265 1270

<210> 110

<211> 1260

<212> PRT

<213> 人工序列

<220>

<223> 重组CoV S多肽

<400> 110

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gly Ser Ala Ser

660 665 670

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala

675 680 685

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn

690 695 700

Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys

705 710 715 720

Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys

725 730 735

Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg

740 745 750

Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val

755 760 765

Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe

770 775 780

Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser

785 790 795 800

Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala

805 810 815

Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala

820 825 830

Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu

835 840 845

Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu

850 855 860

Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala

865 870 875 880

Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile

885 890 895

Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn

900 905 910

Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr

915 920 925

Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln

930 935 940

Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile

945 950 955 960

Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala

965 970 975

Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln

980 985 990

Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser

995 1000 1005

Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln

1010 1015 1020

Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser

1025 1030 1035

Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr

1040 1045 1050

Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile

1055 1060 1065

Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val

1070 1075 1080

Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu

1085 1090 1095

Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys

1100 1105 1110

Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu

1115 1120 1125

Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe

1130 1135 1140

Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly

1145 1150 1155

Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu

1160 1165 1170

Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln

1175 1180 1185

Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile

1190 1195 1200

Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr

1205 1210 1215

Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly

1220 1225 1230

Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser

1235 1240 1245

Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255 1260

<210> 111

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 重组弗林蛋白酶切割位点

<400> 111

Gly Ser Gly Ala

1

<210> 112

<211> 1260

<212> PRT

<213> 人工序列

<220>

<223> 重组SARS-CoV-2多刺突蛋白

<400> 112

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln

660 665 670

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala

675 680 685

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn

690 695 700

Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys

705 710 715 720

Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys

725 730 735

Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg

740 745 750

Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val

755 760 765

Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe

770 775 780

Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser

785 790 795 800

Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala

805 810 815

Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala

820 825 830

Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu

835 840 845

Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu

850 855 860

Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala

865 870 875 880

Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile

885 890 895

Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn

900 905 910

Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr

915 920 925

Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln

930 935 940

Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile

945 950 955 960

Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala

965 970 975

Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln

980 985 990

Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser

995 1000 1005

Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln

1010 1015 1020

Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser

1025 1030 1035

Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr

1040 1045 1050

Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile

1055 1060 1065

Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val

1070 1075 1080

Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu

1085 1090 1095

Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys

1100 1105 1110

Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu

1115 1120 1125

Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe

1130 1135 1140

Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly

1145 1150 1155

Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu

1160 1165 1170

Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln

1175 1180 1185

Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile

1190 1195 1200

Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr

1205 1210 1215

Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly

1220 1225 1230

Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser

1235 1240 1245

Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255 1260

<210> 113

<211> 1260

<212> PRT

<213> 人工序列

<220>

<223> 重组SARS-CoV-2刺突蛋白

<400> 113

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln

660 665 670

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala

675 680 685

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn

690 695 700

Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys

705 710 715 720

Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys

725 730 735

Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg

740 745 750

Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val

755 760 765

Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe

770 775 780

Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser

785 790 795 800

Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala

805 810 815

Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala

820 825 830

Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu

835 840 845

Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu

850 855 860

Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala

865 870 875 880

Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile

885 890 895

Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn

900 905 910

Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr

915 920 925

Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln

930 935 940

Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile

945 950 955 960

Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala

965 970 975

Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln

980 985 990

Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser

995 1000 1005

Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln

1010 1015 1020

Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser

1025 1030 1035

Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr

1040 1045 1050

Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile

1055 1060 1065

Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val

1070 1075 1080

Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu

1085 1090 1095

Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys

1100 1105 1110

Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu

1115 1120 1125

Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe

1130 1135 1140

Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly

1145 1150 1155

Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu

1160 1165 1170

Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln

1175 1180 1185

Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile

1190 1195 1200

Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr

1205 1210 1215

Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly

1220 1225 1230

Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser

1235 1240 1245

Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255 1260

<210> 114

<211> 1257

<212> PRT

<213> 人工序列

<220>

<223> 重组SARS-CoV-2刺突蛋白

<400> 114

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile Ser Gly Thr Asn Gly Thr Lys Arg Phe

50 55 60

Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr

65 70 75 80

Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp

85 90 95

Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val

100 105 110

Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val

115 120 125

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

130 135 140

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

145 150 155 160

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

165 170 175

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

180 185 190

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

195 200 205

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

210 215 220

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

225 230 235 240

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

245 250 255

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

260 265 270

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

275 280 285

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

290 295 300

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

305 310 315 320

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

325 330 335

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

340 345 350

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

355 360 365

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

370 375 380

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

385 390 395 400

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

405 410 415

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

420 425 430

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

435 440 445

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

450 455 460

Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

465 470 475 480

Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

485 490 495

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

500 505 510

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

515 520 525

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

530 535 540

Pro Phe Gln Gln Phe Gly Arg Asp Ile Asp Asp Thr Thr Asp Ala Val

545 550 555 560

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

565 570 575

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

580 585 590

Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile

595 600 605

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

610 615 620

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

625 630 635 640

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

645 650 655

Ser Tyr Gln Thr Gln Thr Asn Ser His Gln Gln Ala Gln Ser Val Ala

660 665 670

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser

675 680 685

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Ile Asn Phe Thr Ile

690 695 700

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

705 710 715 720

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

725 730 735

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

740 745 750

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

755 760 765

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe

770 775 780

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

785 790 795 800

Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly

805 810 815

Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp

820 825 830

Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu

835 840 845

Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly

850 855 860

Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile

865 870 875 880

Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr

885 890 895

Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn

900 905 910

Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala

915 920 925

Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn

930 935 940

Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val

945 950 955 960

Leu Asn Asp Ile Leu Ala Arg Leu Asp Pro Pro Glu Ala Glu Val Gln

965 970 975

Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val

980 985 990

Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu

995 1000 1005

Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg

1010 1015 1020

Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln

1025 1030 1035

Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro

1040 1045 1050

Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp

1055 1060 1065

Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly

1070 1075 1080

Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile

1085 1090 1095

Ile Thr Thr His Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val

1100 1105 1110

Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu

1115 1120 1125

Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His

1130 1135 1140

Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala

1145 1150 1155

Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val

1160 1165 1170

Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly

1175 1180 1185

Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly

1190 1195 1200

Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu

1205 1210 1215

Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser

1220 1225 1230

Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val

1235 1240 1245

Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255

<210> 115

<211> 1260

<212> PRT

<213> 人工序列

<220>

<223> 重组SARS-CoV-2刺突蛋白

<400> 115

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Ala Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Gly Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu His Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Asn Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln

660 665 670

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Val

675 680 685

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn

690 695 700

Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys

705 710 715 720

Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys

725 730 735

Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg

740 745 750

Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val

755 760 765

Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe

770 775 780

Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser

785 790 795 800

Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala

805 810 815

Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala

820 825 830

Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu

835 840 845

Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu

850 855 860

Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala

865 870 875 880

Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile

885 890 895

Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn

900 905 910

Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr

915 920 925

Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln

930 935 940

Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile

945 950 955 960

Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala

965 970 975

Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln

980 985 990

Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser

995 1000 1005

Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln

1010 1015 1020

Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser

1025 1030 1035

Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr

1040 1045 1050

Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile

1055 1060 1065

Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val

1070 1075 1080

Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu

1085 1090 1095

Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys

1100 1105 1110

Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu

1115 1120 1125

Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe

1130 1135 1140

Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly

1145 1150 1155

Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu

1160 1165 1170

Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln

1175 1180 1185

Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile

1190 1195 1200

Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr

1205 1210 1215

Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly

1220 1225 1230

Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser

1235 1240 1245

Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255 1260

<210> 116

<211> 196

<212> PRT

<213> β冠状病毒严重急性呼吸系统综合征冠状病毒 2

<400> 116

Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val

1 5 10 15

Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser

20 25 30

Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val

35 40 45

Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp

50 55 60

Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln

65 70 75 80

Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr

85 90 95

Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly

100 105 110

Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys

115 120 125

Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr

130 135 140

Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser

145 150 155 160

Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val

165 170 175

Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly

180 185 190

Pro Lys Lys Ser

195

<210> 117

<211> 13

<212> PRT

<213> 人工序列

<220>

<223> 信号肽

<400> 117

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ile

1 5 10

<210> 118

<211> 292

<212> PRT

<213> β冠状病毒严重急性呼吸系统综合征冠状病毒 2

<400> 118

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser

290

<210> 119

<211> 83

<212> PRT

<213> β冠状病毒严重急性呼吸系统综合征冠状病毒 2

<400> 119

Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val Leu Tyr

1 5 10 15

Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys

20 25 30

Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln

35 40 45

Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu Val Lys Gln

50 55 60

Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn Asp Ile Leu

65 70 75 80

Ser Arg Leu

<210> 120

<211> 588

<212> PRT

<213> β冠状病毒严重急性呼吸系统综合征冠状病毒 2

<400> 120

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala

1 5 10 15

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn

20 25 30

Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys

35 40 45

Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys

50 55 60

Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg

65 70 75 80

Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val

85 90 95

Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe

100 105 110

Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser

115 120 125

Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala

130 135 140

Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala

145 150 155 160

Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu

165 170 175

Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu

180 185 190

Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala

195 200 205

Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile

210 215 220

Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn

225 230 235 240

Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr

245 250 255

Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln

260 265 270

Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile

275 280 285

Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala

290 295 300

Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln

305 310 315 320

Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser

325 330 335

Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser

340 345 350

Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro

355 360 365

Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro

370 375 380

Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly

385 390 395 400

Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His

405 410 415

Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr

420 425 430

Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val

435 440 445

Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys

450 455 460

Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp

465 470 475 480

Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys

485 490 495

Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu

500 505 510

Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro

515 520 525

Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met

530 535 540

Val Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys

545 550 555 560

Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser

565 570 575

Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

580 585

<210> 121

<211> 672

<212> PRT

<213> β冠状病毒严重急性呼吸系统综合征冠状病毒 2

<400> 121

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg

660 665 670

<210> 122

<211> 140

<212> PRT

<213> β冠状病毒严重急性呼吸系统综合征冠状病毒 2

<400> 122

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

1 5 10 15

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

20 25 30

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

35 40 45

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

50 55 60

Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro

65 70 75 80

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

85 90 95

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

100 105 110

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

115 120 125

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro

130 135 140

<210> 123

<211> 24

<212> PRT

<213> β冠状病毒严重急性呼吸系统综合征冠状病毒 2

<400> 123

Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met

1 5 10 15

Val Thr Ile Met Leu Cys Cys Met

20

<210> 124

<211> 36

<212> PRT

<213> β冠状病毒严重急性呼吸系统综合征冠状病毒 2

<400> 124

Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly Ser Cys

1 5 10 15

Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu Lys Gly Val Lys

20 25 30

Leu His Tyr Thr

35

<210> 125

<211> 51

<212> PRT

<213> β冠状病毒严重急性呼吸系统综合征冠状病毒 2

<400> 125

Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn

1 5 10 15

Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn

20 25 30

Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile

35 40 45

Lys Trp Pro

50

<210> 126

<211> 197

<212> PRT

<213> β冠状病毒严重急性呼吸系统综合征冠状病毒 2

<400> 126

Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg

1 5 10 15

Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val

20 25 30

Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys

35 40 45

Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn

50 55 60

Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile

65 70 75 80

Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro

85 90 95

Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp

100 105 110

Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys

115 120 125

Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln

130 135 140

Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe

145 150 155 160

Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln

165 170 175

Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala

180 185 190

Thr Val Cys Gly Pro

195

<210> 127

<211> 3

<212> PRT

<213> 人工序列

<220>

<223> 无活性的弗林蛋白酶切割位点

<400> 127

Gly Gly Gly

1

<210> 128

<211> 4

<212> PRT

<213> 人工序列

<220>

<223> 无活性的弗林蛋白酶切割位点

<400> 128

Gly Gly Gly Gly

1

<210> 129

<211> 5

<212> PRT

<213> 人工序列

<220>

<223> 无活性的弗林蛋白酶切割位点

<400> 129

Gly Gly Gly Gly Gly

1 5

<210> 130

<211> 1272

<212> PRT

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 130

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Ala

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Gly Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu His Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Asn Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln Ser Val Ala

675 680 685

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Val Glu Asn Ser

690 695 700

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

705 710 715 720

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

725 730 735

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

740 745 750

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

755 760 765

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

770 775 780

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Asn

785 790 795 800

Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser Phe

805 810 815

Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe

820 825 830

Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu

835 840 845

Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu Leu

850 855 860

Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr

865 870 875 880

Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro

885 890 895

Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln

900 905 910

Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser

915 920 925

Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu

930 935 940

Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn Thr

945 950 955 960

Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val Leu

965 970 975

Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln Ile

980 985 990

Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr

995 1000 1005

Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu

1010 1015 1020

Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg

1025 1030 1035

Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln

1040 1045 1050

Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro

1055 1060 1065

Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp

1070 1075 1080

Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly

1085 1090 1095

Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile

1100 1105 1110

Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val

1115 1120 1125

Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu

1130 1135 1140

Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His

1145 1150 1155

Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala

1160 1165 1170

Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val

1175 1180 1185

Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly

1190 1195 1200

Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly

1205 1210 1215

Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu

1220 1225 1230

Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser

1235 1240 1245

Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val

1250 1255 1260

Leu Lys Gly Val Lys Leu His Tyr Thr

1265 1270

<210> 131

<211> 3822

<212> DNA

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 131

atgttcgtct tcctggtgct gctgcctctg gtgtccagcc agtgcgtgaa cctgaccact 60

aggactcagc tgcctcccgc ttacaccaac tcattcacta ggggtgtgta ctacccagac 120

aaggtcttcc gctcctcagt gctgcactca actcaggacc tgttcctgcc tttcttctcc 180

aacgtcacct ggttccacgc tatccacgtg tccggcacca acggcactaa gaggttcgcc 240

aaccccgtgc tgccattcaa cgacggagtc tacttcgctt caaccgagaa gtccaacatc 300

atcagaggat ggatcttcgg caccactctg gacagcaaga ctcagtctct gctgatcgtc 360

aacaacgcta ccaacgtggt catcaaggtg tgcgaattcc agttctgtaa cgaccccttc 420

ctgggtgtct actaccacaa gaacaacaag tcatggatgg agtccgaatt ccgcgtctac 480

tccagcgcta acaactgcac tttcgagtac gtgtcccagc cattcctgat ggacctggaa 540

ggaaagcagg gtaacttcaa gaacctgcgc gagttcgtgt tcaagaacat cgacggctac 600

ttcaagattt acagcaagca cacccccatc aacctggtcc gtggcctgcc acagggattc 660

tctgctctgg agcccctggt ggacctgcca atcggcatca acatcaccag gttccagact 720

ctgcacgccc tgcacagatc ctacctgact cccggagact cttcatccgg ttggaccgct 780

ggcgctgccg cttactacgt gggttacctg cagccacgca ccttcctgct gaagtacaac 840

gaaaacggca ccatcactga cgccgtcgac tgcgctctgg acccactgtc agaaaccaag 900

tgcactctga agtccttcac cgtggagaag ggaatctacc agacttcaaa cttccgcgtg 960

cagcctaccg aatccatcgt ccgtttccca aacatcacta acctgtgccc tttcggagag 1020

gtcttcaacg ccacccgctt cgcttccgtg tacgcctgga accgcaagcg tatctcaaac 1080

tgcgtcgctg actactccgt gctgtacaac tcagcctcct tcagcacctt caagtgctac 1140

ggagtgtcac ccactaagct gaacgacctg tgcttcacca acgtctacgc tgactccttc 1200

gtgatccgcg gtgacgaggt ccgtcagatc gctccaggcc agactggaaa catcgccgac 1260

tacaactaca agctgcctga cgacttcacc ggttgcgtca tcgcctggaa cagcaacaac 1320

ctggactcta aagtgggtgg caactacaac tacctgtaca ggctgttcag aaagtcaaac 1380

ctgaagccct tcgagcgcga catcagcact gaaatctacc aggctggttc taccccatgc 1440

aacggtgtca agggcttcaa ctgctacttc cctctgcagt cctacggctt ccagcccact 1500

tacggtgtgg gctaccagcc ataccgtgtg gtcgtgctga gcttcgagct gctgcacgcc 1560

cctgctactg tgtgcggacc caagaagtct accaacctgg tcaagaacaa gtgcgtgaac 1620

ttcaacttca acggtctgac cggaactggt gtcctgaccg agagcaacaa gaagttcctg 1680

cctttccagc agttcggtcg cgacatcgct gacaccactg acgccgtgcg tgaccctcag 1740

accctggaaa tcctggacat cactccctgc tcattcggag gtgtctccgt gatcactccc 1800

ggaaccaaca cttctaacca ggtcgctgtg ctgtaccagg gtgtgaactg caccgaagtc 1860

ccagtggcca tccacgctga ccagctgacc cctacttgga gggtgtactc caccggcagc 1920

aacgtcttcc agactagagc tggatgcctg atcggtgccg agcacgtcaa caacagctac 1980

gaatgcgaca tccctatcgg cgctggaatc tgcgcctctt accagaccca gactaacagc 2040

ccccagcagg ctcagtctgt ggcctctcag tcaatcatcg cctacaccat gtcactgggt 2100

gtcgaaaact ccgtggctta ctctaacaac tcaatcgcca tccctaccaa cttcactatc 2160

agcgtgacca ctgagatcct gcccgtcagc atgaccaaga cttctgtgga ctgcactatg 2220

tacatctgcg gcgacagcac cgaatgctct aacctgctgc tgcagtacgg ttctttctgc 2280

acccagctga accgcgctct gactggcatc gccgtggagc aggacaagaa cactcaggaa 2340

gtcttcgctc aggtgaagca aatctacaag accccaccta tcaaggactt cggcggattc 2400

aacttctccc agatcctgcc agacccttcc aagcccagca agaggtcttt catcgaggac 2460

ctgctgttca acaaggtcac tctggccgac gctggtttca tcaagcagta cggcgactgc 2520

ctgggagaca tcgctgctcg cgacctgatc tgcgcccaga agttcaacgg cctgaccgtg 2580

ctgcccccac tgctgactga cgaaatgatc gctcagtaca ctagcgccct gctggctggc 2640

accatcactt caggttggac tttcggtgct ggtgctgctc tgcagatccc attcgctatg 2700

cagatggcct accgtttcaa cggaatcggt gtcacccaga acgtgctgta cgagaaccag 2760

aagctgatcg ctaaccagtt caactcagcc atcggcaaga tccaggacag cctgagctct 2820

actgcctctg ctctgggaaa gctgcaggac gtcgtgaacc agaacgccca ggctctgaac 2880

accctggtca agcagctgtc atccaacttc ggcgccatca gctctgtgct gaacgacatc 2940

ctgtccaggc tggaccctcc cgaggctgaa gtccagatcg acaggctgat cactggaaga 3000

ctgcagtcac tgcagaccta cgtgactcag cagctgatcc gcgccgctga aatccgtgcc 3060

tccgctaacc tggccgctac caagatgagc gagtgcgtcc tgggccagtc taagagagtg 3120

gacttctgcg gcaagggata ccacctgatg tcattcccac agtccgcccc tcacggagtc 3180

gtgttcctgc acgtcaccta cgtgcccgct caggagaaga acttcaccac tgccccagct 3240

atctgccacg acggcaaggc tcacttccct cgcgaaggag tcttcgtgag caacggcacc 3300

cactggttcg tcactcagcg taacttctac gagccccaga tcatcaccac tgacaacact 3360

ttcgtgtctg gcaactgcga cgtggtcatc ggtatcgtca acaacaccgt gtacgaccct 3420

ctgcagcccg agctggactc attcaaggaa gagctggaca agtacttcaa gaaccacacc 3480

tcccctgacg tcgacctggg tgacatctca ggcatcaacg cctccgtcgt gaacatccag 3540

aaggagatcg accgcctgaa cgaagtggct aagaacctga acgaaagcct gatcgacctg 3600

caggagctgg gcaagtacga acagtacatc aagtggccat ggtacatctg gctgggattc 3660

atcgctggcc tcatcgctat cgtgatggtg accatcatgc tgtgctgcat gacttcatgc 3720

tgctcctgcc tgaagggttg ctgcagctgc ggctcttgct gcaagttcga cgaggacgac 3780

tctgaacctg tcctgaaggg agtgaagctg cactacacct aa 3822

<210> 132

<211> 1259

<212> PRT

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 132

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Ala Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Gly Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu His Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Asn Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln

660 665 670

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Val

675 680 685

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn

690 695 700

Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys

705 710 715 720

Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys

725 730 735

Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg

740 745 750

Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val

755 760 765

Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe

770 775 780

Gly Gly Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys

785 790 795 800

Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp

805 810 815

Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala

820 825 830

Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro

835 840 845

Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu

850 855 860

Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu

865 870 875 880

Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly

885 890 895

Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln

900 905 910

Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala

915 920 925

Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala

930 935 940

Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser

945 950 955 960

Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu

965 970 975

Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr

980 985 990

Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala

995 1000 1005

Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser

1010 1015 1020

Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe

1025 1030 1035

Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr

1040 1045 1050

Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys

1055 1060 1065

His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser

1070 1075 1080

Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro

1085 1090 1095

Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp

1100 1105 1110

Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln

1115 1120 1125

Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys

1130 1135 1140

Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile

1145 1150 1155

Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn

1160 1165 1170

Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu

1175 1180 1185

Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp

1190 1195 1200

Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile

1205 1210 1215

Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys

1220 1225 1230

Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu

1235 1240 1245

Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255

<210> 133

<211> 1258

<212> PRT

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 133

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gly Gly Ser Val

660 665 670

Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn

675 680 685

Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr

690 695 700

Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser

705 710 715 720

Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn

725 730 735

Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu

740 745 750

Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala

755 760 765

Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly

770 775 780

Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg

785 790 795 800

Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala

805 810 815

Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg

820 825 830

Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro

835 840 845

Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala

850 855 860

Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln

865 870 875 880

Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val

885 890 895

Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe

900 905 910

Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser

915 920 925

Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu

930 935 940

Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser

945 950 955 960

Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val

965 970 975

Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr

980 985 990

Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn

995 1000 1005

Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

1010 1015 1020

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro

1025 1030 1035

Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val

1040 1045 1050

Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

1055 1060 1065

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

1070 1075 1080

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln

1085 1090 1095

Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val

1100 1105 1110

Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro

1115 1120 1125

Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn

1130 1135 1140

His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn

1145 1150 1155

Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu

1160 1165 1170

Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu

1175 1180 1185

Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu

1190 1195 1200

Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met

1205 1210 1215

Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys

1220 1225 1230

Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro

1235 1240 1245

Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255

<210> 134

<211> 1271

<212> PRT

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 134

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gly Gly Ser Val Ala Ser Gln

675 680 685

Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser Val Ala

690 695 700

Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile Ser Val

705 710 715 720

Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys

725 730 735

Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu

740 745 750

Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr Gly Ile

755 760 765

Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys

770 775 780

Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe Asn Phe

785 790 795 800

Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser Phe Ile

805 810 815

Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile

820 825 830

Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile

835 840 845

Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu Leu Thr

850 855 860

Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile

865 870 875 880

Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe

885 890 895

Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn

900 905 910

Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala

915 920 925

Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly

930 935 940

Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu

945 950 955 960

Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn

965 970 975

Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln Ile Asp

980 985 990

Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln

995 1000 1005

Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala

1010 1015 1020

Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val

1025 1030 1035

Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ser

1040 1045 1050

Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala

1055 1060 1065

Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly

1070 1075 1080

Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr

1085 1090 1095

His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile

1100 1105 1110

Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile

1115 1120 1125

Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu

1130 1135 1140

Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr

1145 1150 1155

Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser

1160 1165 1170

Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala

1175 1180 1185

Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys

1190 1195 1200

Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe

1205 1210 1215

Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu Cys

1220 1225 1230

Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys

1235 1240 1245

Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu

1250 1255 1260

Lys Gly Val Lys Leu His Tyr Thr

1265 1270

<210> 135

<211> 3813

<212> PRT

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 135

Ala Thr Gly Thr Thr Cys Gly Thr Cys Thr Thr Cys Cys Thr Gly Gly

1 5 10 15

Thr Gly Cys Thr Gly Cys Thr Gly Cys Cys Thr Cys Thr Gly Gly Thr

20 25 30

Gly Thr Cys Cys Ala Gly Cys Cys Ala Gly Thr Gly Cys Gly Thr Gly

35 40 45

Ala Ala Cys Cys Thr Gly Ala Cys Cys Ala Cys Thr Ala Gly Gly Ala

50 55 60

Cys Thr Cys Ala Gly Cys Thr Gly Cys Cys Thr Cys Cys Cys Gly Cys

65 70 75 80

Thr Thr Ala Cys Ala Cys Cys Ala Ala Cys Ala Gly Cys Thr Thr Cys

85 90 95

Ala Cys Thr Ala Gly Gly Gly Gly Thr Gly Thr Cys Thr Ala Cys Thr

100 105 110

Ala Cys Cys Cys Thr Gly Ala Cys Ala Ala Gly Gly Thr Cys Thr Thr

115 120 125

Cys Cys Gly Cys Thr Cys Cys Thr Cys Ala Gly Thr Gly Cys Thr Gly

130 135 140

Cys Ala Cys Thr Cys Thr Ala Cys Thr Cys Ala Gly Gly Ala Cys Cys

145 150 155 160

Thr Gly Thr Thr Cys Cys Thr Gly Cys Cys Cys Thr Thr Cys Thr Thr

165 170 175

Cys Thr Cys Ala Ala Ala Cys Gly Thr Gly Ala Cys Cys Thr Gly Gly

180 185 190

Thr Thr Cys Cys Ala Cys Gly Cys Cys Ala Thr Cys Thr Cys Cys Gly

195 200 205

Gly Cys Ala Cys Cys Ala Ala Cys Gly Gly Cys Ala Cys Thr Ala Ala

210 215 220

Gly Ala Gly Gly Thr Thr Cys Gly Ala Cys Ala Ala Cys Cys Cys Cys

225 230 235 240

Gly Thr Gly Cys Thr Gly Cys Cys Ala Thr Thr Cys Ala Ala Cys Gly

245 250 255

Ala Cys Gly Gly Cys Gly Thr Cys Thr Ala Cys Thr Thr Cys Gly Cys

260 265 270

Thr Thr Cys Ala Ala Cys Cys Gly Ala Gly Ala Ala Gly Thr Cys Cys

275 280 285

Ala Ala Cys Ala Thr Cys Ala Thr Cys Ala Gly Ala Gly Gly Ala Thr

290 295 300

Gly Gly Ala Thr Cys Thr Thr Cys Gly Gly Cys Ala Cys Cys Ala Cys

305 310 315 320

Thr Cys Thr Gly Gly Ala Cys Ala Gly Cys Ala Ala Gly Ala Cys Thr

325 330 335

Cys Ala Gly Thr Cys Thr Cys Thr Gly Cys Thr Gly Ala Thr Cys Gly

340 345 350

Thr Cys Ala Ala Cys Ala Ala Cys Gly Cys Cys Ala Cys Cys Ala Ala

355 360 365

Cys Gly Thr Gly Gly Thr Cys Ala Thr Cys Ala Ala Gly Gly Thr Gly

370 375 380

Thr Gly Cys Gly Ala Ala Thr Thr Cys Cys Ala Gly Thr Thr Cys Thr

385 390 395 400

Gly Cys Ala Ala Cys Gly Ala Cys Cys Cys Ala Thr Thr Cys Cys Thr

405 410 415

Gly Gly Gly Thr Gly Thr Gly Thr Ala Cys Cys Ala Cys Ala Ala Gly

420 425 430

Ala Ala Cys Ala Ala Cys Ala Ala Gly Thr Cys Ala Thr Gly Gly Ala

435 440 445

Thr Gly Gly Ala Gly Thr Cys Cys Gly Ala Ala Thr Thr Cys Cys Gly

450 455 460

Cys Gly Thr Cys Thr Ala Cys Thr Cys Cys Ala Gly Cys Gly Cys Thr

465 470 475 480

Ala Ala Cys Ala Ala Cys Thr Gly Cys Ala Cys Thr Thr Thr Cys Gly

485 490 495

Ala Gly Thr Ala Cys Gly Thr Gly Ala Gly Cys Cys Ala Gly Cys Cys

500 505 510

Thr Thr Thr Cys Cys Thr Gly Ala Thr Gly Gly Ala Cys Cys Thr Gly

515 520 525

Gly Ala Ala Gly Gly Ala Ala Ala Gly Cys Ala Gly Gly Gly Thr Ala

530 535 540

Ala Cys Thr Thr Cys Ala Ala Gly Ala Ala Cys Cys Thr Gly Cys Gly

545 550 555 560

Cys Gly Ala Gly Thr Thr Cys Gly Thr Gly Thr Thr Cys Ala Ala Gly

565 570 575

Ala Ala Cys Ala Thr Cys Gly Ala Cys Gly Gly Ala Thr Ala Cys Thr

580 585 590

Thr Cys Ala Ala Gly Ala Thr Thr Thr Ala Cys Ala Gly Cys Ala Ala

595 600 605

Gly Cys Ala Cys Ala Cys Cys Cys Cys Cys Ala Thr Cys Ala Ala Cys

610 615 620

Cys Thr Gly Gly Thr Cys Cys Gly Thr Gly Ala Cys Cys Thr Gly Cys

625 630 635 640

Cys Ala Cys Ala Gly Gly Gly Thr Thr Thr Cys Thr Cys Thr Gly Cys

645 650 655

Thr Cys Thr Gly Gly Ala Gly Cys Cys Cys Cys Thr Gly Gly Thr Gly

660 665 670

Gly Ala Cys Cys Thr Gly Cys Cys Ala Ala Thr Cys Gly Gly Cys Ala

675 680 685

Thr Cys Ala Ala Cys Ala Thr Cys Ala Cys Cys Ala Gly Gly Thr Thr

690 695 700

Cys Cys Ala Gly Ala Cys Thr Cys Thr Gly Cys Thr Gly Gly Cys Thr

705 710 715 720

Cys Thr Gly Cys Ala Cys Cys Gly Cys Thr Cys Cys Thr Ala Cys Cys

725 730 735

Thr Gly Ala Cys Thr Cys Cys Ala Gly Gly Thr Gly Ala Cys Thr Cys

740 745 750

Ala Thr Cys Thr Thr Cys Thr Gly Gly Ala Thr Gly Gly Ala Cys Thr

755 760 765

Gly Cys Thr Gly Gly Ala Gly Cys Thr Gly Cys Thr Gly Cys Thr Thr

770 775 780

Ala Cys Thr Ala Cys Gly Thr Gly Gly Gly Thr Thr Ala Cys Cys Thr

785 790 795 800

Gly Cys Ala Gly Cys Cys Thr Cys Gly Cys Ala Cys Cys Thr Thr Cys

805 810 815

Cys Thr Gly Cys Thr Gly Ala Ala Gly Thr Ala Cys Ala Ala Cys Gly

820 825 830

Ala Ala Ala Ala Cys Gly Gly Cys Ala Cys Cys Ala Thr Cys Ala Cys

835 840 845

Thr Gly Ala Cys Gly Cys Cys Gly Thr Cys Gly Ala Cys Thr Gly Cys

850 855 860

Gly Cys Thr Cys Thr Gly Gly Ala Cys Cys Cys Thr Cys Thr Gly Thr

865 870 875 880

Cys Ala Gly Ala Ala Ala Cys Cys Ala Ala Gly Thr Gly Cys Ala Cys

885 890 895

Thr Cys Thr Gly Ala Ala Gly Thr Cys Cys Thr Thr Cys Ala Cys Cys

900 905 910

Gly Thr Gly Gly Ala Gly Ala Ala Gly Gly Gly Thr Ala Thr Cys Thr

915 920 925

Ala Cys Cys Ala Gly Ala Cys Thr Ala Gly Cys Ala Ala Cys Thr Thr

930 935 940

Cys Cys Gly Cys Gly Thr Gly Cys Ala Gly Cys Cys Cys Ala Cys Cys

945 950 955 960

Gly Ala Ala Thr Cys Thr Ala Thr Cys Gly Thr Cys Cys Gly Thr Thr

965 970 975

Thr Cys Cys Cys Ala Ala Ala Cys Ala Thr Cys Ala Cys Thr Ala Ala

980 985 990

Cys Cys Thr Gly Thr Gly Cys Cys Cys Thr Thr Thr Cys Gly Gly Cys

995 1000 1005

Gly Ala Gly Gly Thr Cys Thr Thr Cys Ala Ala Cys Gly Cys Cys

1010 1015 1020

Ala Cys Cys Cys Gly Cys Thr Thr Cys Gly Cys Thr Thr Cys Cys

1025 1030 1035

Gly Thr Gly Thr Ala Cys Gly Cys Cys Thr Gly Gly Ala Ala Cys

1040 1045 1050

Cys Gly Cys Ala Ala Gly Cys Gly Thr Ala Thr Cys Thr Cys Ala

1055 1060 1065

Ala Ala Cys Thr Gly Cys Gly Thr Cys Gly Cys Thr Gly Ala Cys

1070 1075 1080

Thr Ala Cys Thr Cys Cys Gly Thr Gly Cys Thr Gly Thr Ala Cys

1085 1090 1095

Ala Ala Cys Thr Cys Ala Gly Cys Cys Thr Cys Cys Thr Thr Cys

1100 1105 1110

Ala Gly Cys Ala Cys Cys Thr Thr Cys Ala Ala Gly Thr Gly Cys

1115 1120 1125

Thr Ala Cys Gly Gly Thr Gly Thr Gly Thr Cys Ala Cys Cys Cys

1130 1135 1140

Ala Cys Thr Ala Ala Gly Cys Thr Gly Ala Ala Cys Gly Ala Cys

1145 1150 1155

Cys Thr Gly Thr Gly Cys Thr Thr Cys Ala Cys Cys Ala Ala Cys

1160 1165 1170

Gly Thr Cys Thr Ala Cys Gly Cys Cys Gly Ala Cys Thr Cys Cys

1175 1180 1185

Thr Thr Cys Gly Thr Gly Ala Thr Cys Cys Gly Cys Gly Gly Cys

1190 1195 1200

Gly Ala Cys Gly Ala Gly Gly Thr Cys Cys Gly Thr Cys Ala Gly

1205 1210 1215

Ala Thr Cys Gly Cys Thr Cys Cys Ala Gly Gly Cys Cys Ala Gly

1220 1225 1230

Ala Cys Thr Gly Gly Ala Ala Ala Gly Ala Thr Cys Gly Cys Cys

1235 1240 1245

Gly Ala Cys Thr Ala Cys Ala Ala Cys Thr Ala Cys Ala Ala Gly

1250 1255 1260

Cys Thr Gly Cys Cys Thr Gly Ala Cys Gly Ala Cys Thr Thr Cys

1265 1270 1275

Ala Cys Cys Gly Gly Cys Thr Gly Cys Gly Thr Cys Ala Thr Cys

1280 1285 1290

Gly Cys Thr Thr Gly Gly Ala Ala Cys Ala Gly Cys Ala Ala Cys

1295 1300 1305

Ala Ala Cys Cys Thr Gly Gly Ala Cys Thr Cys Thr Ala Ala Ala

1310 1315 1320

Gly Thr Gly Gly Gly Thr Gly Gly Cys Ala Ala Cys Thr Ala Cys

1325 1330 1335

Ala Ala Cys Thr Ala Cys Cys Thr Gly Thr Ala Cys Ala Gly Gly

1340 1345 1350

Cys Thr Gly Thr Thr Cys Ala Gly Ala Ala Ala Gly Ala Gly Cys

1355 1360 1365

Ala Ala Cys Cys Thr Gly Ala Ala Gly Cys Cys Cys Thr Thr Cys

1370 1375 1380

Gly Ala Gly Cys Gly Cys Gly Ala Cys Ala Thr Cys Ala Gly Cys

1385 1390 1395

Ala Cys Thr Gly Ala Ala Ala Thr Cys Thr Ala Cys Cys Ala Gly

1400 1405 1410

Gly Cys Thr Gly Gly Cys Thr Cys Thr Ala Cys Cys Cys Cys Ala

1415 1420 1425

Thr Gly Cys Ala Ala Cys Gly Gly Thr Gly Thr Cys Gly Ala Gly

1430 1435 1440

Gly Gly Cys Thr Thr Cys Ala Ala Cys Thr Gly Cys Thr Ala Cys

1445 1450 1455

Thr Thr Cys Cys Cys Thr Cys Thr Gly Cys Ala Gly Thr Cys Thr

1460 1465 1470

Thr Ala Cys Gly Gly Ala Thr Thr Cys Cys Ala Gly Cys Cys Cys

1475 1480 1485

Ala Cys Thr Thr Ala Cys Gly Gly Thr Gly Thr Gly Gly Gly Cys

1490 1495 1500

Thr Ala Cys Cys Ala Gly Cys Cys Ala Thr Ala Cys Cys Gly Thr

1505 1510 1515

Gly Thr Gly Gly Thr Cys Gly Thr Gly Cys Thr Gly Ala Gly Cys

1520 1525 1530

Thr Thr Cys Gly Ala Ala Cys Thr Gly Cys Thr Gly Cys Ala Cys

1535 1540 1545

Gly Cys Cys Cys Cys Thr Gly Cys Thr Ala Cys Thr Gly Thr Gly

1550 1555 1560

Thr Gly Cys Gly Gly Ala Cys Cys Cys Ala Ala Gly Ala Ala Gly

1565 1570 1575

Thr Cys Thr Ala Cys Cys Ala Ala Cys Cys Thr Gly Gly Thr Cys

1580 1585 1590

Ala Ala Gly Ala Ala Cys Ala Ala Gly Thr Gly Cys Gly Thr Gly

1595 1600 1605

Ala Ala Cys Thr Thr Cys Ala Ala Cys Thr Thr Cys Ala Ala Cys

1610 1615 1620

Gly Gly Thr Cys Thr Gly Ala Cys Cys Gly Gly Ala Ala Cys Thr

1625 1630 1635

Gly Gly Thr Gly Thr Cys Cys Thr Gly Ala Cys Cys Gly Ala Gly

1640 1645 1650

Ala Gly Cys Ala Ala Cys Ala Ala Gly Ala Ala Gly Thr Thr Cys

1655 1660 1665

Cys Thr Gly Cys Cys Cys Thr Thr Cys Cys Ala Gly Cys Ala Gly

1670 1675 1680

Thr Thr Cys Gly Gly Thr Cys Gly Cys Gly Ala Cys Ala Thr Cys

1685 1690 1695

Gly Ala Cys Gly Ala Cys Ala Cys Cys Ala Cys Thr Gly Ala Cys

1700 1705 1710

Gly Cys Cys Gly Thr Gly Cys Gly Thr Gly Ala Cys Cys Cys Thr

1715 1720 1725

Cys Ala Gly Ala Cys Cys Cys Thr Gly Gly Ala Ala Ala Thr Cys

1730 1735 1740

Cys Thr Gly Gly Ala Cys Ala Thr Cys Ala Cys Thr Cys Cys Cys

1745 1750 1755

Thr Gly Cys Thr Cys Ala Thr Thr Cys Gly Gly Ala Gly Gly Thr

1760 1765 1770

Gly Thr Cys Thr Cys Cys Gly Thr Gly Ala Thr Cys Ala Cys Thr

1775 1780 1785

Cys Cys Ala Gly Gly Ala Ala Cys Cys Ala Ala Cys Ala Cys Thr

1790 1795 1800

Thr Cys Thr Ala Ala Cys Cys Ala Gly Gly Thr Cys Gly Cys Thr

1805 1810 1815

Gly Thr Gly Cys Thr Gly Thr Ala Cys Cys Ala Gly Gly Gly Thr

1820 1825 1830

Gly Thr Cys Ala Ala Cys Thr Gly Cys Ala Cys Cys Gly Ala Gly

1835 1840 1845

Gly Thr Cys Cys Cys Ala Gly Thr Gly Gly Cys Cys Ala Thr Cys

1850 1855 1860

Cys Ala Cys Gly Cys Thr Gly Ala Cys Cys Ala Gly Cys Thr Gly

1865 1870 1875

Ala Cys Cys Cys Cys Thr Ala Cys Thr Thr Gly Gly Ala Gly Gly

1880 1885 1890

Gly Thr Gly Thr Ala Cys Thr Cys Cys Ala Cys Cys Gly Gly Cys

1895 1900 1905

Ala Gly Cys Ala Ala Cys Gly Thr Gly Thr Thr Cys Cys Ala Gly

1910 1915 1920

Ala Cys Thr Ala Gly Ala Gly Cys Thr Gly Gly Thr Thr Gly Cys

1925 1930 1935

Cys Thr Gly Ala Thr Cys Gly Gly Cys Gly Cys Cys Gly Ala Gly

1940 1945 1950

Cys Ala Cys Gly Thr Gly Ala Ala Cys Ala Ala Cys Ala Gly Cys

1955 1960 1965

Thr Ala Cys Gly Ala Ala Thr Gly Cys Gly Ala Cys Ala Thr Cys

1970 1975 1980

Cys Cys Ala Ala Thr Cys Gly Gly Cys Gly Cys Thr Gly Gly Ala

1985 1990 1995

Ala Thr Cys Thr Gly Cys Gly Cys Cys Thr Cys Thr Thr Ala Cys

2000 2005 2010

Cys Ala Gly Ala Cys Cys Cys Ala Gly Ala Cys Thr Ala Ala Cys

2015 2020 2025

Ala Gly Cys Cys Ala Cys Cys Ala Gly Cys Ala Gly Gly Cys Thr

2030 2035 2040

Cys Ala Gly Thr Cys Thr Gly Thr Cys Gly Cys Cys Thr Cys Thr

2045 2050 2055

Cys Ala Gly Thr Cys Ala Ala Thr Cys Ala Thr Cys Gly Cys Thr

2060 2065 2070

Thr Ala Cys Ala Cys Thr Ala Thr Gly Thr Cys Ala Cys Thr Gly

2075 2080 2085

Gly Gly Cys Gly Cys Thr Gly Ala Ala Ala Ala Cys Thr Cys Cys

2090 2095 2100

Gly Thr Gly Gly Cys Cys Thr Ala Cys Thr Cys Thr Ala Ala Cys

2105 2110 2115

Ala Ala Cys Thr Cys Ala Ala Thr Cys Gly Cys Cys Ala Thr Cys

2120 2125 2130

Cys Cys Thr Ala Thr Cys Ala Ala Cys Thr Thr Cys Ala Cys Cys

2135 2140 2145

Ala Thr Cys Thr Cys Ala Gly Thr Gly Ala Cys Cys Ala Cys Thr

2150 2155 2160

Gly Ala Gly Ala Thr Cys Cys Thr Gly Cys Cys Cys Gly Thr Cys

2165 2170 2175

Ala Gly Cys Ala Thr Gly Ala Cys Cys Ala Ala Gly Ala Cys Thr

2180 2185 2190

Thr Cys Thr Gly Thr Gly Gly Ala Cys Thr Gly Cys Ala Cys Thr

2195 2200 2205

Ala Thr Gly Thr Ala Cys Ala Thr Cys Thr Gly Cys Gly Gly Ala

2210 2215 2220

Gly Ala Cys Ala Gly Cys Ala Cys Cys Gly Ala Ala Thr Gly Cys

2225 2230 2235

Thr Cys Thr Ala Ala Cys Cys Thr Gly Cys Thr Gly Cys Thr Gly

2240 2245 2250

Cys Ala Gly Thr Ala Cys Gly Gly Cys Thr Cys Cys Thr Thr Cys

2255 2260 2265

Thr Gly Cys Ala Cys Cys Cys Ala Gly Cys Thr Gly Ala Ala Cys

2270 2275 2280

Cys Gly Cys Gly Cys Thr Cys Thr Gly Ala Cys Thr Gly Gly Ala

2285 2290 2295

Ala Thr Cys Gly Cys Cys Gly Thr Gly Gly Ala Gly Cys Ala Gly

2300 2305 2310

Gly Ala Cys Ala Ala Gly Ala Ala Cys Ala Cys Thr Cys Ala Gly

2315 2320 2325

Gly Ala Ala Gly Thr Cys Thr Thr Cys Gly Cys Thr Cys Ala Gly

2330 2335 2340

Gly Thr Gly Ala Ala Gly Cys Ala Ala Ala Thr Cys Thr Ala Cys

2345 2350 2355

Ala Ala Gly Ala Cys Cys Cys Cys Ala Cys Cys Thr Ala Thr Cys

2360 2365 2370

Ala Ala Gly Gly Ala Cys Thr Thr Cys Gly Gly Cys Gly Gly Ala

2375 2380 2385

Thr Thr Cys Ala Ala Cys Thr Thr Cys Thr Cys Cys Cys Ala Gly

2390 2395 2400

Ala Thr Cys Cys Thr Gly Cys Cys Ala Gly Ala Cys Cys Cys Thr

2405 2410 2415

Thr Cys Cys Ala Ala Gly Cys Cys Ala Ala Gly Cys Ala Ala Gly

2420 2425 2430

Ala Gly Gly Thr Cys Thr Thr Thr Cys Ala Thr Cys Gly Ala Gly

2435 2440 2445

Gly Ala Cys Cys Thr Gly Cys Thr Gly Thr Thr Cys Ala Ala Cys

2450 2455 2460

Ala Ala Gly Gly Thr Cys Ala Cys Cys Cys Thr Gly Gly Cys Cys

2465 2470 2475

Gly Ala Cys Gly Cys Thr Gly Gly Cys Thr Thr Cys Ala Thr Cys

2480 2485 2490

Ala Ala Gly Cys Ala Gly Thr Ala Cys Gly Gly Ala Gly Ala Cys

2495 2500 2505

Thr Gly Cys Cys Thr Gly Gly Gly Thr Gly Ala Cys Ala Thr Cys

2510 2515 2520

Gly Cys Cys Gly Cys Thr Cys Gly Cys Gly Ala Cys Cys Thr Gly

2525 2530 2535

Ala Thr Cys Thr Gly Cys Gly Cys Thr Cys Ala Gly Ala Ala Gly

2540 2545 2550

Thr Thr Cys Ala Ala Cys Gly Gly Ala Cys Thr Gly Ala Cys Cys

2555 2560 2565

Gly Thr Gly Cys Thr Gly Cys Cys Cys Cys Cys Ala Cys Thr Gly

2570 2575 2580

Cys Thr Gly Ala Cys Thr Gly Ala Cys Gly Ala Ala Ala Thr Gly

2585 2590 2595

Ala Thr Cys Gly Cys Cys Cys Ala Gly Thr Ala Cys Ala Cys Thr

2600 2605 2610

Thr Cys Ala Gly Cys Cys Cys Thr Gly Cys Thr Gly Gly Cys Thr

2615 2620 2625

Gly Gly Ala Ala Cys Cys Ala Thr Cys Ala Cys Thr Thr Cys Thr

2630 2635 2640

Gly Gly Thr Thr Gly Gly Ala Cys Cys Thr Thr Cys Gly Gly Thr

2645 2650 2655

Gly Cys Thr Gly Gly Thr Gly Cys Thr Gly Cys Thr Cys Thr Gly

2660 2665 2670

Cys Ala Gly Ala Thr Cys Cys Cys Thr Thr Thr Cys Gly Cys Thr

2675 2680 2685

Ala Thr Gly Cys Ala Gly Ala Thr Gly Gly Cys Cys Thr Ala Cys

2690 2695 2700

Cys Gly Thr Thr Thr Cys Ala Ala Cys Gly Gly Ala Ala Thr Cys

2705 2710 2715

Gly Gly Thr Gly Thr Cys Ala Cys Cys Cys Ala Gly Ala Ala Cys

2720 2725 2730

Gly Thr Gly Cys Thr Gly Thr Ala Cys Gly Ala Gly Ala Ala Cys

2735 2740 2745

Cys Ala Gly Ala Ala Gly Cys Thr Gly Ala Thr Cys Gly Cys Thr

2750 2755 2760

Ala Ala Cys Cys Ala Gly Thr Thr Cys Ala Ala Cys Thr Cys Cys

2765 2770 2775

Gly Cys Cys Ala Thr Cys Gly Gly Cys Ala Ala Gly Ala Thr Cys

2780 2785 2790

Cys Ala Gly Gly Ala Cys Ala Gly Cys Cys Thr Gly Ala Gly Cys

2795 2800 2805

Thr Cys Thr Ala Cys Thr Gly Cys Cys Thr Cys Thr Gly Cys Thr

2810 2815 2820

Cys Thr Gly Gly Gly Ala Ala Ala Gly Cys Thr Gly Cys Ala Gly

2825 2830 2835

Gly Ala Cys Gly Thr Cys Gly Thr Gly Ala Ala Cys Cys Ala Gly

2840 2845 2850

Ala Ala Cys Gly Cys Cys Cys Ala Gly Gly Cys Thr Cys Thr Gly

2855 2860 2865

Ala Ala Cys Ala Cys Cys Cys Thr Gly Gly Thr Cys Ala Ala Gly

2870 2875 2880

Cys Ala Gly Cys Thr Gly Thr Cys Ala Thr Cys Cys Ala Ala Cys

2885 2890 2895

Thr Thr Cys Gly Gly Ala Gly Cys Cys Ala Thr Cys Ala Gly Cys

2900 2905 2910

Thr Cys Thr Gly Thr Gly Cys Thr Gly Ala Ala Cys Gly Ala Cys

2915 2920 2925

Ala Thr Cys Cys Thr Cys Gly Cys Thr Ala Gly Gly Cys Thr Gly

2930 2935 2940

Gly Ala Cys Cys Cys Thr Cys Cys Cys Gly Ala Gly Gly Cys Thr

2945 2950 2955

Gly Ala Ala Gly Thr Cys Cys Ala Gly Ala Thr Cys Gly Ala Cys

2960 2965 2970

Ala Gly Gly Cys Thr Gly Ala Thr Cys Ala Cys Thr Gly Gly Thr

2975 2980 2985

Ala Gly Ala Cys Thr Gly Cys Ala Gly Thr Cys Ala Cys Thr Gly

2990 2995 3000

Cys Ala Gly Ala Cys Cys Thr Ala Cys Gly Thr Gly Ala Cys Thr

3005 3010 3015

Cys Ala Gly Cys Ala Gly Cys Thr Gly Ala Thr Cys Cys Gly Cys

3020 3025 3030

Gly Cys Cys Gly Cys Thr Gly Ala Ala Ala Thr Cys Cys Gly Thr

3035 3040 3045

Gly Cys Cys Thr Cys Cys Gly Cys Thr Ala Ala Cys Cys Thr Gly

3050 3055 3060

Gly Cys Cys Gly Cys Thr Ala Cys Cys Ala Ala Gly Ala Thr Gly

3065 3070 3075

Ala Gly Cys Gly Ala Gly Thr Gly Cys Gly Thr Cys Cys Thr Gly

3080 3085 3090

Gly Gly Ala Cys Ala Gly Thr Cys Thr Ala Ala Gly Ala Gly Ala

3095 3100 3105

Gly Thr Gly Gly Ala Cys Thr Thr Cys Thr Gly Cys Gly Gly Cys

3110 3115 3120

Ala Ala Gly Gly Gly Ala Thr Ala Cys Cys Ala Cys Cys Thr Gly

3125 3130 3135

Ala Thr Gly Thr Cys Ala Thr Thr Cys Cys Cys Ala Cys Ala Gly

3140 3145 3150

Thr Cys Cys Gly Cys Thr Cys Cys Thr Cys Ala Cys Gly Gly Thr

3155 3160 3165

Gly Thr Cys Gly Thr Gly Thr Thr Cys Cys Thr Gly Cys Ala Cys

3170 3175 3180

Gly Thr Cys Ala Cys Cys Thr Ala Cys Gly Thr Gly Cys Cys Cys

3185 3190 3195

Gly Cys Cys Cys Ala Gly Gly Ala Gly Ala Ala Gly Ala Ala Cys

3200 3205 3210

Thr Thr Cys Ala Cys Cys Ala Cys Thr Gly Cys Cys Cys Cys Ala

3215 3220 3225

Gly Cys Thr Ala Thr Cys Thr Gly Cys Cys Ala Cys Gly Ala Cys

3230 3235 3240

Gly Gly Cys Ala Ala Gly Gly Cys Thr Cys Ala Cys Thr Thr Cys

3245 3250 3255

Cys Cys Cys Cys Gly Cys Gly Ala Ala Gly Gly Thr Gly Thr Cys

3260 3265 3270

Thr Thr Cys Gly Thr Gly Thr Cys Ala Ala Ala Cys Gly Gly Cys

3275 3280 3285

Ala Cys Cys Cys Ala Cys Thr Gly Gly Thr Thr Cys Gly Thr Cys

3290 3295 3300

Ala Cys Thr Cys Ala Gly Cys Gly Thr Ala Ala Cys Thr Thr Cys

3305 3310 3315

Thr Ala Cys Gly Ala Gly Cys Cys Ala Cys Ala Gly Ala Thr Cys

3320 3325 3330

Ala Thr Cys Ala Cys Cys Ala Cys Thr Cys Ala Cys Ala Ala Cys

3335 3340 3345

Ala Cys Thr Thr Thr Cys Gly Thr Gly Thr Cys Cys Gly Gly Ala

3350 3355 3360

Ala Ala Cys Thr Gly Cys Gly Ala Cys Gly Thr Gly Gly Thr Cys

3365 3370 3375

Ala Thr Cys Gly Gly Thr Ala Thr Cys Gly Thr Cys Ala Ala Cys

3380 3385 3390

Ala Ala Cys Ala Cys Cys Gly Thr Gly Thr Ala Cys Gly Ala Cys

3395 3400 3405

Cys Cys Thr Cys Thr Gly Cys Ala Gly Cys Cys Cys Gly Ala Gly

3410 3415 3420

Cys Thr Gly Gly Ala Cys Thr Cys Ala Thr Thr Cys Ala Ala Gly

3425 3430 3435

Gly Ala Ala Gly Ala Gly Cys Thr Gly Gly Ala Cys Ala Ala Gly

3440 3445 3450

Thr Ala Cys Thr Thr Cys Ala Ala Gly Ala Ala Cys Cys Ala Cys

3455 3460 3465

Ala Cys Cys Thr Cys Cys Cys Cys Thr Gly Ala Cys Gly Thr Cys

3470 3475 3480

Gly Ala Cys Cys Thr Gly Gly Gly Cys Gly Ala Cys Ala Thr Cys

3485 3490 3495

Thr Cys Ala Gly Gly Ala Ala Thr Cys Ala Ala Cys Gly Cys Thr

3500 3505 3510

Thr Cys Cys Gly Thr Cys Gly Thr Gly Ala Ala Cys Ala Thr Cys

3515 3520 3525

Cys Ala Gly Ala Ala Gly Gly Ala Gly Ala Thr Cys Gly Ala Cys

3530 3535 3540

Cys Gly Cys Cys Thr Gly Ala Ala Cys Gly Ala Ala Gly Thr Gly

3545 3550 3555

Gly Cys Cys Ala Ala Gly Ala Ala Cys Cys Thr Gly Ala Ala Cys

3560 3565 3570

Gly Ala Ala Thr Cys Ala Cys Thr Gly Ala Thr Cys Gly Ala Cys

3575 3580 3585

Cys Thr Gly Cys Ala Gly Gly Ala Gly Cys Thr Gly Gly Gly Cys

3590 3595 3600

Ala Ala Gly Thr Ala Cys Gly Ala Ala Cys Ala Gly Thr Ala Cys

3605 3610 3615

Ala Thr Cys Ala Ala Gly Thr Gly Gly Cys Cys Thr Thr Gly Gly

3620 3625 3630

Thr Ala Cys Ala Thr Cys Thr Gly Gly Cys Thr Gly Gly Gly Thr

3635 3640 3645

Thr Thr Cys Ala Thr Cys Gly Cys Thr Gly Gly Cys Cys Thr Cys

3650 3655 3660

Ala Thr Cys Gly Cys Thr Ala Thr Cys Gly Thr Gly Ala Thr Gly

3665 3670 3675

Gly Thr Gly Ala Cys Cys Ala Thr Cys Ala Thr Gly Cys Thr Gly

3680 3685 3690

Thr Gly Cys Thr Gly Cys Ala Thr Gly Ala Cys Thr Thr Cys Ala

3695 3700 3705

Thr Gly Cys Thr Gly Cys Thr Cys Cys Thr Gly Cys Cys Thr Gly

3710 3715 3720

Ala Ala Gly Gly Gly Cys Thr Gly Cys Thr Gly Cys Ala Gly Cys

3725 3730 3735

Thr Gly Cys Gly Gly Ala Thr Cys Thr Thr Gly Cys Thr Gly Cys

3740 3745 3750

Ala Ala Gly Thr Thr Cys Gly Ala Cys Gly Ala Gly Gly Ala Cys

3755 3760 3765

Gly Ala Cys Thr Cys Cys Gly Ala Ala Cys Cys Cys Gly Thr Cys

3770 3775 3780

Cys Thr Gly Ala Ala Gly Gly Gly Ala Gly Thr Gly Ala Ala Gly

3785 3790 3795

Cys Thr Gly Cys Ala Cys Thr Ala Cys Ala Cys Cys Thr Ala Ala

3800 3805 3810

<210> 136

<211> 1270

<212> PRT

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 136

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp Asn Pro

65 70 75 80

Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu Lys Ser

85 90 95

Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser Lys Thr

100 105 110

Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile Lys Val

115 120 125

Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr His Lys

130 135 140

Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr Ser Ser Ala

145 150 155 160

Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu Met Asp Leu

165 170 175

Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe Val Phe Lys

180 185 190

Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr Pro Ile Asn

195 200 205

Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu Pro Leu Val

210 215 220

Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr Leu Leu Ala

225 230 235 240

Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser Gly Trp Thr

245 250 255

Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro Arg Thr Phe

260 265 270

Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala Val Asp Cys

275 280 285

Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser Phe Thr

290 295 300

Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln Pro Thr

305 310 315 320

Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly

325 330 335

Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg

340 345 350

Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser

355 360 365

Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu

370 375 380

Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg

385 390 395 400

Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala

405 410 415

Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala

420 425 430

Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr

435 440 445

Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp

450 455 460

Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val

465 470 475 480

Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro

485 490 495

Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe

500 505 510

Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr

515 520 525

Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn Gly Leu Thr

530 535 540

Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro Phe Gln

545 550 555 560

Gln Phe Gly Arg Asp Ile Asp Asp Thr Thr Asp Ala Val Arg Asp Pro

565 570 575

Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly Val

580 585 590

Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val Ala Val Leu

595 600 605

Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile His Ala Asp

610 615 620

Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser Asn Val Phe

625 630 635 640

Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val Asn Asn Ser

645 650 655

Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser Tyr Gln

660 665 670

Thr Gln Thr Asn Ser His Gln Gln Ala Gln Ser Val Ala Ser Gln Ser

675 680 685

Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser Val Ala Tyr

690 695 700

Ser Asn Asn Ser Ile Ala Ile Pro Ile Asn Phe Thr Ile Ser Val Thr

705 710 715 720

Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys Thr

725 730 735

Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu Gln

740 745 750

Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr Gly Ile Ala

755 760 765

Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys Gln

770 775 780

Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe Asn Phe Ser

785 790 795 800

Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser Phe Ile Glu

805 810 815

Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile Lys

820 825 830

Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile Cys

835 840 845

Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu Leu Thr Asp

850 855 860

Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile Thr

865 870 875 880

Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala

885 890 895

Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val

900 905 910

Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile

915 920 925

Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys

930 935 940

Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu Val

945 950 955 960

Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn Asp

965 970 975

Ile Leu Ala Arg Leu Asp Pro Pro Glu Ala Glu Val Gln Ile Asp Arg

980 985 990

Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln

995 1000 1005

Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala

1010 1015 1020

Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val Asp

1025 1030 1035

Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ser Ala

1040 1045 1050

Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala Gln

1055 1060 1065

Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys

1070 1075 1080

Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His

1085 1090 1095

Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr

1100 1105 1110

Thr His Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly

1115 1120 1125

Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp

1130 1135 1140

Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser

1145 1150 1155

Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val

1160 1165 1170

Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys

1175 1180 1185

Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr

1190 1195 1200

Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile

1205 1210 1215

Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu Cys Cys

1220 1225 1230

Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly

1235 1240 1245

Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu Lys

1250 1255 1260

Gly Val Lys Leu His Tyr Thr

1265 1270

<210> 137

<211> 1268

<212> PRT

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 137

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp Asn Pro

65 70 75 80

Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu Lys Ser

85 90 95

Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser Lys Thr

100 105 110

Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile Lys Val

115 120 125

Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr His Lys

130 135 140

Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr Ser Ser Ala

145 150 155 160

Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu Met Asp Leu

165 170 175

Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe Val Phe Lys

180 185 190

Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr Pro Ile Asn

195 200 205

Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu Pro Leu Val

210 215 220

Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr Leu Leu Ala

225 230 235 240

Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser Gly Trp Thr

245 250 255

Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro Arg Thr Phe

260 265 270

Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala Val Asp Cys

275 280 285

Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser Phe Thr

290 295 300

Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln Pro Thr

305 310 315 320

Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly

325 330 335

Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg

340 345 350

Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser

355 360 365

Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu

370 375 380

Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg

385 390 395 400

Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala

405 410 415

Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala

420 425 430

Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr

435 440 445

Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp

450 455 460

Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val

465 470 475 480

Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro

485 490 495

Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe

500 505 510

Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr

515 520 525

Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn Gly Leu Thr

530 535 540

Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro Phe Gln

545 550 555 560

Gln Phe Gly Arg Asp Ile Asp Asp Thr Thr Asp Ala Val Arg Asp Pro

565 570 575

Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly Val

580 585 590

Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val Ala Val Leu

595 600 605

Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile His Ala Asp

610 615 620

Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser Asn Val Phe

625 630 635 640

Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val Asn Asn Ser

645 650 655

Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser Tyr Gln

660 665 670

Thr Gln Thr Asn Ser His Gly Gly Ser Val Ala Ser Gln Ser Ile Ile

675 680 685

Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser Val Ala Tyr Ser Asn

690 695 700

Asn Ser Ile Ala Ile Pro Ile Asn Phe Thr Ile Ser Val Thr Thr Glu

705 710 715 720

Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys Thr Met Tyr

725 730 735

Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu Gln Tyr Gly

740 745 750

Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr Gly Ile Ala Val Glu

755 760 765

Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys Gln Ile Tyr

770 775 780

Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe Asn Phe Ser Gln Ile

785 790 795 800

Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser Phe Ile Glu Asp Leu

805 810 815

Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile Lys Gln Tyr

820 825 830

Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile Cys Ala Gln

835 840 845

Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu Leu Thr Asp Glu Met

850 855 860

Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile Thr Ser Gly

865 870 875 880

Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala Met Gln

885 890 895

Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val Leu Tyr

900 905 910

Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys

915 920 925

Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln

930 935 940

Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu Val Lys Gln

945 950 955 960

Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn Asp Ile Leu

965 970 975

Ala Arg Leu Asp Pro Pro Glu Ala Glu Val Gln Ile Asp Arg Leu Ile

980 985 990

Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile

995 1000 1005

Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala Thr Lys

1010 1015 1020

Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val Asp Phe Cys

1025 1030 1035

Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ser Ala Pro His

1040 1045 1050

Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala Gln Glu Lys

1055 1060 1065

Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys Ala His

1070 1075 1080

Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His Trp Phe

1085 1090 1095

Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr His

1100 1105 1110

Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val

1115 1120 1125

Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe

1130 1135 1140

Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp

1145 1150 1155

Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn

1160 1165 1170

Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu

1175 1180 1185

Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln

1190 1195 1200

Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly

1205 1210 1215

Leu Ile Ala Ile Val Met Val Thr Ile Met Leu Cys Cys Met Thr

1220 1225 1230

Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly Ser Cys

1235 1240 1245

Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu Lys Gly Val

1250 1255 1260

Lys Leu His Tyr Thr

1265

<210> 138

<211> 1255

<212> PRT

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 138

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile Ser Gly Thr Asn Gly Thr Lys Arg Phe

50 55 60

Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr

65 70 75 80

Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp

85 90 95

Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val

100 105 110

Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val

115 120 125

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

130 135 140

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

145 150 155 160

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

165 170 175

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

180 185 190

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

195 200 205

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

210 215 220

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

225 230 235 240

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

245 250 255

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

260 265 270

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

275 280 285

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

290 295 300

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

305 310 315 320

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

325 330 335

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

340 345 350

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

355 360 365

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

370 375 380

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

385 390 395 400

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

405 410 415

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

420 425 430

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

435 440 445

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

450 455 460

Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

465 470 475 480

Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

485 490 495

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

500 505 510

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

515 520 525

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

530 535 540

Pro Phe Gln Gln Phe Gly Arg Asp Ile Asp Asp Thr Thr Asp Ala Val

545 550 555 560

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

565 570 575

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

580 585 590

Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile

595 600 605

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

610 615 620

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

625 630 635 640

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

645 650 655

Ser Tyr Gln Thr Gln Thr Asn Ser His Gly Gly Ser Val Ala Ser Gln

660 665 670

Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser Val Ala

675 680 685

Tyr Ser Asn Asn Ser Ile Ala Ile Pro Ile Asn Phe Thr Ile Ser Val

690 695 700

Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys

705 710 715 720

Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu

725 730 735

Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr Gly Ile

740 745 750

Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys

755 760 765

Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe Asn Phe

770 775 780

Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser Phe Ile

785 790 795 800

Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile

805 810 815

Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile

820 825 830

Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu Leu Thr

835 840 845

Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile

850 855 860

Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe

865 870 875 880

Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn

885 890 895

Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala

900 905 910

Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly

915 920 925

Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu

930 935 940

Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn

945 950 955 960

Asp Ile Leu Ala Arg Leu Asp Pro Pro Glu Ala Glu Val Gln Ile Asp

965 970 975

Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln

980 985 990

Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala

995 1000 1005

Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val Asp

1010 1015 1020

Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ser Ala

1025 1030 1035

Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala Gln

1040 1045 1050

Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys

1055 1060 1065

Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His

1070 1075 1080

Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr

1085 1090 1095

Thr His Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly

1100 1105 1110

Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp

1115 1120 1125

Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser

1130 1135 1140

Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val

1145 1150 1155

Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys

1160 1165 1170

Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr

1175 1180 1185

Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile

1190 1195 1200

Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu Cys Cys

1205 1210 1215

Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly

1220 1225 1230

Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu Lys

1235 1240 1245

Gly Val Lys Leu His Tyr Thr

1250 1255

<210> 139

<211> 3807

<212> DNA

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 139

atgttcgtct tcctggtgct gctgcctctg gtgtccagcc agtgcgtgaa cctgaccact 60

aggactcagc tgcctcccgc ttacaccaac agcttcacta ggggtgtcta ctaccctgac 120

aaggtcttcc gctcctcagt gctgcactct actcaggacc tgttcctgcc cttcttctca 180

aacgtgacct ggttccacgc catctccggc accaacggca ctaagaggtt cgacaacccc 240

gtgctgccat tcaacgacgg cgtctacttc gcttcaaccg agaagtccaa catcatcaga 300

ggatggatct tcggcaccac tctggacagc aagactcagt ctctgctgat cgtcaacaac 360

gccaccaacg tggtcatcaa ggtgtgcgaa ttccagttct gcaacgaccc attcctgggt 420

gtgtaccaca agaacaacaa gtcatggatg gagtccgaat tccgcgtcta ctccagcgct 480

aacaactgca ctttcgagta cgtgagccag cctttcctga tggacctgga aggaaagcag 540

ggtaacttca agaacctgcg cgagttcgtg ttcaagaaca tcgacggata cttcaagatt 600

tacagcaagc acacccccat caacctggtc cgtgacctgc cacagggttt ctctgctctg 660

gagcccctgg tggacctgcc aatcggcatc aacatcacca ggttccagac tctgctggct 720

ctgcaccgct cctacctgac tccaggtgac tcatcttctg gatggactgc tggagctgct 780

gcttactacg tgggttacct gcagcctcgc accttcctgc tgaagtacaa cgaaaacggc 840

accatcactg acgccgtcga ctgcgctctg gaccctctgt cagaaaccaa gtgcactctg 900

aagtccttca ccgtggagaa gggtatctac cagactagca acttccgcgt gcagcccacc 960

gaatctatcg tccgtttccc aaacatcact aacctgtgcc ctttcggcga ggtcttcaac 1020

gccacccgct tcgcttccgt gtacgcctgg aaccgcaagc gtatctcaaa ctgcgtcgct 1080

gactactccg tgctgtacaa ctcagcctcc ttcagcacct tcaagtgcta cggtgtgtca 1140

cccactaagc tgaacgacct gtgcttcacc aacgtctacg ccgactcctt cgtgatccgc 1200

ggcgacgagg tccgtcagat cgctccaggc cagactggaa agatcgccga ctacaactac 1260

aagctgcctg acgacttcac cggctgcgtc atcgcttgga acagcaacaa cctggactct 1320

aaagtgggtg gcaactacaa ctacctgtac aggctgttca gaaagagcaa cctgaagccc 1380

ttcgagcgcg acatcagcac tgaaatctac caggctggct ctaccccatg caacggtgtc 1440

gagggcttca actgctactt ccctctgcag tcttacggat tccagcccac ttacggtgtg 1500

ggctaccagc cataccgtgt ggtcgtgctg agcttcgaac tgctgcacgc ccctgctact 1560

gtgtgcggac ccaagaagtc taccaacctg gtcaagaaca agtgcgtgaa cttcaacttc 1620

aacggtctga ccggaactgg tgtcctgacc gagagcaaca agaagttcct gcccttccag 1680

cagttcggtc gcgacatcga cgacaccact gacgccgtgc gtgaccctca gaccctggaa 1740

atcctggaca tcactccctg ctcattcgga ggtgtctccg tgatcactcc aggaaccaac 1800

acttctaacc aggtcgctgt gctgtaccag ggtgtcaact gcaccgaggt cccagtggcc 1860

atccacgctg accagctgac ccctacttgg agggtgtact ccaccggcag caacgtgttc 1920

cagactagag ctggttgcct gatcggcgcc gagcacgtga acaacagcta cgaatgcgac 1980

atcccaatcg gcgctggaat ctgcgcctct taccagaccc agactaacag ccacggtggt 2040

tctgtcgcct ctcagtcaat catcgcttac actatgtcac tgggcgctga aaactccgtg 2100

gcctactcta acaactcaat cgccatccct atcaacttca ccatctcagt gaccactgag 2160

atcctgcccg tcagcatgac caagacttct gtggactgca ctatgtacat ctgcggagac 2220

agcaccgaat gctctaacct gctgctgcag tacggctcct tctgcaccca gctgaaccgc 2280

gctctgactg gaatcgccgt ggagcaggac aagaacactc aggaagtctt cgctcaggtg 2340

aagcaaatct acaagacccc acctatcaag gacttcggcg gattcaactt ctcccagatc 2400

ctgccagacc cttccaagcc aagcaagagg tctttcatcg aggacctgct gttcaacaag 2460

gtcaccctgg ccgacgctgg cttcatcaag cagtacggag actgcctggg tgacatcgcc 2520

gctcgcgacc tgatctgcgc tcagaagttc aacggactga ccgtgctgcc cccactgctg 2580

actgacgaaa tgatcgccca gtacacttca gccctgctgg ctggaaccat cacttctggt 2640

tggaccttcg gtgctggtgc tgctctgcag atccctttcg ctatgcagat ggcctaccgt 2700

ttcaacggaa tcggtgtcac ccagaacgtg ctgtacgaga accagaagct gatcgctaac 2760

cagttcaact ccgccatcgg caagatccag gacagcctga gctctactgc ctctgctctg 2820

ggaaagctgc aggacgtcgt gaaccagaac gcccaggctc tgaacaccct ggtcaagcag 2880

ctgtcatcca acttcggagc catcagctct gtgctgaacg acatcctcgc taggctggac 2940

cctcccgagg ctgaagtcca gatcgacagg ctgatcactg gtagactgca gtcactgcag 3000

acctacgtga ctcagcagct gatccgcgcc gctgaaatcc gtgcctccgc taacctggcc 3060

gctaccaaga tgagcgagtg cgtcctggga cagtctaaga gagtggactt ctgcggcaag 3120

ggataccacc tgatgtcatt cccacagtcc gctcctcacg gtgtcgtgtt cctgcacgtc 3180

acctacgtgc ccgcccagga gaagaacttc accactgccc cagctatctg ccacgacggc 3240

aaggctcact tcccccgcga aggtgtcttc gtgtcaaacg gcacccactg gttcgtcact 3300

cagcgtaact tctacgagcc acagatcatc accactcaca acactttcgt gtccggaaac 3360

tgcgacgtgg tcatcggtat cgtcaacaac accgtgtacg accctctgca gcccgagctg 3420

gactcattca aggaagagct ggacaagtac ttcaagaacc acacctcccc tgacgtcgac 3480

ctgggcgaca tctcaggaat caacgcttcc gtcgtgaaca tccagaagga gatcgaccgc 3540

ctgaacgaag tggccaagaa cctgaacgaa tcactgatcg acctgcagga gctgggcaag 3600

tacgaacagt acatcaagtg gccttggtac atctggctgg gtttcatcgc tggcctcatc 3660

gctatcgtga tggtgaccat catgctgtgc tgcatgactt catgctgctc ctgcctgaag 3720

ggctgctgca gctgcggatc ttgctgcaag ttcgacgagg acgactccga acccgtcctg 3780

aagggagtga agctgcacta cacctaa 3807

<210> 140

<211> 1270

<212> PRT

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 140

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Ala

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Gly Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu His Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Asn Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gly Gly Ser Val Ala Ser Gln

675 680 685

Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Val Glu Asn Ser Val Ala

690 695 700

Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile Ser Val

705 710 715 720

Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys

725 730 735

Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu

740 745 750

Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr Gly Ile

755 760 765

Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys

770 775 780

Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Asn Phe Ser

785 790 795 800

Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser Phe Ile Glu

805 810 815

Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile Lys

820 825 830

Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile Cys

835 840 845

Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu Leu Thr Asp

850 855 860

Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile Thr

865 870 875 880

Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala

885 890 895

Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val

900 905 910

Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile

915 920 925

Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys

930 935 940

Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu Val

945 950 955 960

Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn Asp

965 970 975

Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln Ile Asp Arg

980 985 990

Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln

995 1000 1005

Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala

1010 1015 1020

Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val Asp

1025 1030 1035

Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ser Ala

1040 1045 1050

Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala Gln

1055 1060 1065

Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys

1070 1075 1080

Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His

1085 1090 1095

Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr

1100 1105 1110

Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly

1115 1120 1125

Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp

1130 1135 1140

Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser

1145 1150 1155

Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val

1160 1165 1170

Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys

1175 1180 1185

Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr

1190 1195 1200

Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile

1205 1210 1215

Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu Cys Cys

1220 1225 1230

Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly

1235 1240 1245

Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu Lys

1250 1255 1260

Gly Val Lys Leu His Tyr Thr

1265 1270

<210> 141

<211> 1257

<212> PRT

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 141

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Ala Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Gly Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu His Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Asn Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gly Gly Ser Val

660 665 670

Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Val Glu Asn

675 680 685

Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr

690 695 700

Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser

705 710 715 720

Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn

725 730 735

Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu

740 745 750

Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala

755 760 765

Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly

770 775 780

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

785 790 795 800

Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly

805 810 815

Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp

820 825 830

Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu

835 840 845

Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly

850 855 860

Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile

865 870 875 880

Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr

885 890 895

Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn

900 905 910

Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala

915 920 925

Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn

930 935 940

Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val

945 950 955 960

Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln

965 970 975

Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val

980 985 990

Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu

995 1000 1005

Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg

1010 1015 1020

Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln

1025 1030 1035

Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro

1040 1045 1050

Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp

1055 1060 1065

Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly

1070 1075 1080

Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile

1085 1090 1095

Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val

1100 1105 1110

Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu

1115 1120 1125

Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His

1130 1135 1140

Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala

1145 1150 1155

Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val

1160 1165 1170

Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly

1175 1180 1185

Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly

1190 1195 1200

Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu

1205 1210 1215

Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser

1220 1225 1230

Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val

1235 1240 1245

Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255

<210> 142

<211> 3816

<212> DNA

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 142

atgttcgtct tcctggtgct gctgcctctg gtgtccagcc agtgcgtgaa cctgaccact 60

aggactcagc tgcctcccgc ttacaccaac tcattcacta ggggtgtgta ctacccagac 120

aaggtcttcc gctcctcagt gctgcactca actcaggacc tgttcctgcc tttcttctcc 180

aacgtcacct ggttccacgc tatccacgtg tccggcacca acggcactaa gaggttcgcc 240

aaccccgtgc tgccattcaa cgacggagtc tacttcgctt caaccgagaa gtccaacatc 300

atcagaggat ggatcttcgg caccactctg gacagcaaga ctcagtctct gctgatcgtc 360

aacaacgcta ccaacgtggt catcaaggtg tgcgaattcc agttctgtaa cgaccccttc 420

ctgggtgtct actaccacaa gaacaacaag tcatggatgg agtccgaatt ccgcgtctac 480

tccagcgcta acaactgcac tttcgagtac gtgtcccagc cattcctgat ggacctggaa 540

ggaaagcagg gtaacttcaa gaacctgcgc gagttcgtgt tcaagaacat cgacggctac 600

ttcaagattt acagcaagca cacccccatc aacctggtcc gtggcctgcc acagggattc 660

tctgctctgg agcccctggt ggacctgcca atcggcatca acatcaccag gttccagact 720

ctgcacgccc tgcacagatc ctacctgact cccggagact cttcatccgg ttggaccgct 780

ggcgctgccg cttactacgt gggttacctg cagccacgca ccttcctgct gaagtacaac 840

gaaaacggca ccatcactga cgccgtcgac tgcgctctgg acccactgtc agaaaccaag 900

tgcactctga agtccttcac cgtggagaag ggaatctacc agacttcaaa cttccgcgtg 960

cagcctaccg aatccatcgt ccgtttccca aacatcacta acctgtgccc tttcggagag 1020

gtcttcaacg ccacccgctt cgcttccgtg tacgcctgga accgcaagcg tatctcaaac 1080

tgcgtcgctg actactccgt gctgtacaac tcagcctcct tcagcacctt caagtgctac 1140

ggagtgtcac ccactaagct gaacgacctg tgcttcacca acgtctacgc tgactccttc 1200

gtgatccgcg gtgacgaggt ccgtcagatc gctccaggcc agactggaaa catcgccgac 1260

tacaactaca agctgcctga cgacttcacc ggttgcgtca tcgcctggaa cagcaacaac 1320

ctggactcta aagtgggtgg caactacaac tacctgtaca ggctgttcag aaagtcaaac 1380

ctgaagccct tcgagcgcga catcagcact gaaatctacc aggctggttc taccccatgc 1440

aacggtgtca agggcttcaa ctgctacttc cctctgcagt cctacggctt ccagcccact 1500

tacggtgtgg gctaccagcc ataccgtgtg gtcgtgctga gcttcgagct gctgcacgcc 1560

cctgctactg tgtgcggacc caagaagtct accaacctgg tcaagaacaa gtgcgtgaac 1620

ttcaacttca acggtctgac cggaactggt gtcctgaccg agagcaacaa gaagttcctg 1680

cctttccagc agttcggtcg cgacatcgct gacaccactg acgccgtgcg tgaccctcag 1740

accctggaaa tcctggacat cactccctgc tcattcggag gtgtctccgt gatcactccc 1800

ggaaccaaca cttctaacca ggtcgctgtg ctgtaccagg gtgtgaactg caccgaagtc 1860

ccagtggcca tccacgctga ccagctgacc cctacttgga gggtgtactc caccggcagc 1920

aacgtcttcc agactagagc tggatgcctg atcggtgccg agcacgtcaa caacagctac 1980

gaatgcgaca tccctatcgg cgctggaatc tgcgcctctt accagaccca gactaacagc 2040

ccaggtggtt ctgtggcctc tcagtcaatc atcgcctaca ccatgtcact gggtgtcgaa 2100

aactccgtgg cttactctaa caactcaatc gccatcccta ccaacttcac tatcagcgtg 2160

accactgaga tcctgcccgt cagcatgacc aagacttctg tggactgcac tatgtacatc 2220

tgcggcgaca gcaccgaatg ctctaacctg ctgctgcagt acggttcttt ctgcacccag 2280

ctgaaccgcg ctctgactgg catcgccgtg gagcaggaca agaacactca ggaagtcttc 2340

gctcaggtga agcaaatcta caagacccca cctatcaagg acttcggcgg attcaacttc 2400

tcccagatcc tgccagaccc ttccaagccc agcaagaggt ctttcatcga ggacctgctg 2460

ttcaacaagg tcactctggc cgacgctggt ttcatcaagc agtacggcga ctgcctggga 2520

gacatcgctg ctcgcgacct gatctgcgcc cagaagttca acggcctgac cgtgctgccc 2580

ccactgctga ctgacgaaat gatcgctcag tacactagcg ccctgctggc tggcaccatc 2640

acttcaggtt ggactttcgg tgctggtgct gctctgcaga tcccattcgc tatgcagatg 2700

gcctaccgtt tcaacggaat cggtgtcacc cagaacgtgc tgtacgagaa ccagaagctg 2760

atcgctaacc agttcaactc agccatcggc aagatccagg acagcctgag ctctactgcc 2820

tctgctctgg gaaagctgca ggacgtcgtg aaccagaacg cccaggctct gaacaccctg 2880

gtcaagcagc tgtcatccaa cttcggcgcc atcagctctg tgctgaacga catcctgtcc 2940

aggctggacc ctcccgaggc tgaagtccag atcgacaggc tgatcactgg aagactgcag 3000

tcactgcaga cctacgtgac tcagcagctg atccgcgccg ctgaaatccg tgcctccgct 3060

aacctggccg ctaccaagat gagcgagtgc gtcctgggcc agtctaagag agtggacttc 3120

tgcggcaagg gataccacct gatgtcattc ccacagtccg cccctcacgg agtcgtgttc 3180

ctgcacgtca cctacgtgcc cgctcaggag aagaacttca ccactgcccc agctatctgc 3240

cacgacggca aggctcactt ccctcgcgaa ggagtcttcg tgagcaacgg cacccactgg 3300

ttcgtcactc agcgtaactt ctacgagccc cagatcatca ccactgacaa cactttcgtg 3360

tctggcaact gcgacgtggt catcggtatc gtcaacaaca ccgtgtacga ccctctgcag 3420

cccgagctgg actcattcaa ggaagagctg gacaagtact tcaagaacca cacctcccct 3480

gacgtcgacc tgggtgacat ctcaggcatc aacgcctccg tcgtgaacat ccagaaggag 3540

atcgaccgcc tgaacgaagt ggctaagaac ctgaacgaaa gcctgatcga cctgcaggag 3600

ctgggcaagt acgaacagta catcaagtgg ccatggtaca tctggctggg attcatcgct 3660

ggcctcatcg ctatcgtgat ggtgaccatc atgctgtgct gcatgacttc atgctgctcc 3720

tgcctgaagg gttgctgcag ctgcggctct tgctgcaagt tcgacgagga cgactctgaa 3780

cctgtcctga agggagtgaa gctgcactac acctaa 3816

<210> 143

<211> 1269

<212> PRT

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 143

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Phe Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Ala

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Gly Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser Gly Trp Thr

245 250 255

Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro Arg Thr Phe

260 265 270

Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala Val Asp Cys

275 280 285

Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser Phe Thr

290 295 300

Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln Pro Thr

305 310 315 320

Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly

325 330 335

Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg

340 345 350

Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser

355 360 365

Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu

370 375 380

Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg

385 390 395 400

Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Asn Ile Ala

405 410 415

Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala

420 425 430

Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr

435 440 445

Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp

450 455 460

Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val

465 470 475 480

Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro

485 490 495

Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe

500 505 510

Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr

515 520 525

Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn Gly Leu Thr

530 535 540

Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro Phe Gln

545 550 555 560

Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val Arg Asp Pro

565 570 575

Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly Val

580 585 590

Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val Ala Val Leu

595 600 605

Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile His Ala Asp

610 615 620

Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser Asn Val Phe

625 630 635 640

Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val Asn Asn Ser

645 650 655

Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser Tyr Gln

660 665 670

Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln Ser Val Ala Ser Gln Ser

675 680 685

Ile Ile Ala Tyr Thr Met Ser Leu Gly Val Glu Asn Ser Val Ala Tyr

690 695 700

Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile Ser Val Thr

705 710 715 720

Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys Thr

725 730 735

Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu Gln

740 745 750

Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr Gly Ile Ala

755 760 765

Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys Gln

770 775 780

Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Asn Phe Ser Gln

785 790 795 800

Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser Phe Ile Glu Asp

805 810 815

Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile Lys Gln

820 825 830

Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile Cys Ala

835 840 845

Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu Leu Thr Asp Glu

850 855 860

Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile Thr Ser

865 870 875 880

Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala Met

885 890 895

Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val Leu

900 905 910

Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile Gly

915 920 925

Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys Leu

930 935 940

Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu Val Lys

945 950 955 960

Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn Asp Ile

965 970 975

Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln Ile Asp Arg Leu

980 985 990

Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln Leu

995 1000 1005

Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala Thr

1010 1015 1020

Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val Asp Phe

1025 1030 1035

Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ser Ala Pro

1040 1045 1050

His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala Gln Glu

1055 1060 1065

Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys Ala

1070 1075 1080

His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His Trp

1085 1090 1095

Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr

1100 1105 1110

Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile

1115 1120 1125

Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser

1130 1135 1140

Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro

1145 1150 1155

Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val

1160 1165 1170

Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn

1175 1180 1185

Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu

1190 1195 1200

Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile Ala

1205 1210 1215

Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu Cys Cys Met

1220 1225 1230

Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly Ser

1235 1240 1245

Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu Lys Gly

1250 1255 1260

Val Lys Leu His Tyr Thr

1265

<210> 144

<211> 1256

<212> PRT

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 144

Gln Cys Val Asn Phe Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Ala Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Gly Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

225 230 235 240

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

245 250 255

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

260 265 270

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

275 280 285

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

290 295 300

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

305 310 315 320

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

325 330 335

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

340 345 350

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

355 360 365

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

370 375 380

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

385 390 395 400

Asn Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

405 410 415

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

420 425 430

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

435 440 445

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

450 455 460

Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

465 470 475 480

Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

485 490 495

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

500 505 510

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

515 520 525

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

530 535 540

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

545 550 555 560

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

565 570 575

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

580 585 590

Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile

595 600 605

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

610 615 620

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

625 630 635 640

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

645 650 655

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln Ser Val Ala

660 665 670

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Val Glu Asn Ser

675 680 685

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

690 695 700

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

705 710 715 720

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

725 730 735

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

740 745 750

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

755 760 765

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Asn

770 775 780

Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser Phe

785 790 795 800

Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe

805 810 815

Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu

820 825 830

Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu Leu

835 840 845

Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr

850 855 860

Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro

865 870 875 880

Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln

885 890 895

Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser

900 905 910

Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu

915 920 925

Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn Thr

930 935 940

Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val Leu

945 950 955 960

Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln Ile

965 970 975

Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr

980 985 990

Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala

995 1000 1005

Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val

1010 1015 1020

Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ser

1025 1030 1035

Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala

1040 1045 1050

Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly

1055 1060 1065

Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr

1070 1075 1080

His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile

1085 1090 1095

Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile

1100 1105 1110

Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu

1115 1120 1125

Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr

1130 1135 1140

Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser

1145 1150 1155

Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala

1160 1165 1170

Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys

1175 1180 1185

Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe

1190 1195 1200

Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu Cys

1205 1210 1215

Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys

1220 1225 1230

Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu

1235 1240 1245

Lys Gly Val Lys Leu His Tyr Thr

1250 1255

<210> 145

<211> 3812

<212> DNA

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 145

atgttcgtct tcctggtgct gctgcctctg gtgtccagcc agtgcgtgaa cttcaccact 60

aggactcagc tgcctcccgc ttacaccaac tcattcacta ggggtgtgta ctacccagac 120

aaggtcttcc gctcctcagt gctgcactca actcaggacc tgttcctgcc tttcttctcc 180

aacgtcacct ggttccacgc tatccacgtg tccggcacca acggcactaa gaggttcgcc 240

aaccccgtgc tgccattcaa cgacggagtc tacttcgctt caaccgagaa gtccaacatc 300

atcagaggat ggatcttcgg caccactctg gacagcaaga ctcagtctct gctgatcgtc 360

aacaacgcta ccaacgtggt catcaaggtg tgcgaattcc agttctgtaa cgaccccttc 420

ctgggtgtct actaccacaa gaacaacaag tcatggatgg agtccgaatt ccgcgtctac 480

tccagcgcta acaactgcac tttcgagtac gtgtcccagc cattcctgat ggacctggaa 540

ggaaagcagg gtaacttcaa gaacctgcgc gagttcgtgt tcaagaacat cgacggctac 600

ttcaagattt acagcaagca cacccccatc aacctggtcc gtggcctgcc acagggattc 660

tctgctctgg agcccctggt ggacctgcca atcggcatca acatcaccag gttccagact 720

ctgcacagat cctacctgac tcccggagac tcttcatccg gttggaccgc tggcgctgcc 780

gcttactacg tgggttacct gcagccacgc accttcctgc tgaagtacaa cgaaaacggc 840

accatcactg acgccgtcga ctgcgctctg gacccactgt cagaaaccaa gtgcactctg 900

aagtccttca ccgtggagaa gggaatctac cagacttcaa acttccgcgt gcagcctacc 960

gaatccatcg tccgtttccc aaacatcact aacctgtgcc ctttcggaga ggtcttcaac 1020

gccacccgct tcgcttccgt gtacgcctgg aaccgcaagc gtatctcaaa ctgcgtcgct 1080

gactactccg tgctgtacaa ctcagcctcc ttcagcacct tcaagtgcta cggagtgtca 1140

cccactaagc tgaacgacct gtgcttcacc aacgtctacg ctgactcctt cgtgatccgc 1200

ggtgacgagg tccgtcagat cgctccaggc cagactggaa acatcgccga ctacaactac 1260

aagctgcctg acgacttcac cggttgcgtc atcgcctgga acagcaacaa cctggactct 1320

aaagtgggtg gcaactacaa ctacctgtac aggctgttca gaaagtcaaa cctgaagccc 1380

ttcgagcgcg acatcagcac tgaaatctac caggctggtt ctaccccatg caacggtgtc 1440

aagggcttca actgctactt ccctctgcag tcctacggct tccagcccac ttacggtgtg 1500

ggctaccagc cataccgtgt ggtcgtgctg agcttcgagc tgctgcacgc ccctgctact 1560

gtgtgcggac ccaagaagtc taccaacctg gtcaagaaca agtgcgtgaa cttcaacttc 1620

aacggtctga ccggaactgg tgtcctgacc gagagcaaca agaagttcct gcctttccag 1680

cagttcggtc gcgacatcgc tgacaccact gacgccgtgc gtgaccctca gaccctggaa 1740

atcctggaca tcactccctg ctcattcgga ggtgtctccg tgatcactcc cggaaccaac 1800

acttctaacc aggtcgctgt gctgtaccag ggtgtgaact gcaccgaagt cccagtggcc 1860

atccacgctg accagctgac ccctacttgg agggtgtact ccaccggcag caacgtcttc 1920

cagactagag ctggatgcct gatcggtgcc gagcacgtca acaacagcta cgaatgcgac 1980

atccctatcg gcgctggaat ctgcgcctct taccagaccc agactaacag cccccagcag 2040

gctcagtctg tggcctctca gtcaatcatc gcctacacca tgtcactggg tgtcgaaaac 2100

tccgtggctt actctaacaa ctcaatcgcc atccctacca acttcactat cagcgtgacc 2160

actgagatcc tgcccgtcag catgaccaag acttctgtgg actgcactat gtacatctgc 2220

ggcgacagca ccgaatgctc taacctgctg ctgcagtacg gttctttctg cacccagctg 2280

aaccgcgctc tgactggcat cgccgtggag caggacaaga acactcagga agtcttcgct 2340

caggtgaagc aaatctacaa gaccccacct atcaaggact tcggcggatt caacttctcc 2400

cagatcctgc cagacccttc caagcccagc aagaggtctt tcatcgagga cctgctgttc 2460

aacaaggtca ctctggccga cgctggtttc atcaagcagt acggcgactg cctgggagac 2520

atcgctgctc gcgacctgat ctgcgcccag aagttcaacg gcctgaccgt gctgccccca 2580

ctgctgactg acgaaatgat cgctcagtac actagcgccc tgctggctgg caccatcact 2640

tcaggttgga ctttcggtgc tggtgctgct ctgcagatcc cattcgctat gcagatggcc 2700

taccgtttca acggaatcgg tgtcacccag aacgtgctgt acgagaacca gaagctgatc 2760

gctaaccagt tcaactcagc catcggcaag atccaggaca gcctgagctc tactgcctct 2820

gctctgggaa agctgcagga cgtcgtgaac cagaacgccc aggctctgaa caccctggtc 2880

aagcagctgt catccaactt cggcgccatc agctctgtgc tgaacgacat cctgtccagg 2940

ctggaccctc ccgaggctga agtccagatc gacaggctga tcactggaag actgcagtca 3000

ctgcagacct acgtgactca gcagctgatc cgcgccgctg aaatccgtgc ctccgctaac 3060

ctggccgcta ccaagatgag cgagtgcgtc ctgggccagt ctaagagagt ggacttctgc 3120

ggcaagggat accacctgat gtcattccca cagtccgccc ctcacggagt cgtgttcctg 3180

cacgtcacct acgtgcccgc tcaggagaag aacttcacca ctgccccagc tatctgccac 3240

gacggcaagg ctcacttccc tcgcgaagga gtcttcgtga gcaacggcac ccactggttc 3300

gtcactcagc gtaacttcta cgagccccag atcatcacca ctgacaacac tttcgtgtct 3360

ggcaactgcg acgtggtcat cggtatcgtc aacaacaccg tgtacgaccc tctgcagccc 3420

gagctggact cattcaagga agagctggac aagtacttca agaaccacac ctcccctgac 3480

gtcgacctgg gtgacatctc aggcatcaac gcctccgtcg tgaacatcca gaaggagatc 3540

gaccgcctga acgaagtggc taagaacctg aacgaaagcc tgatcgacct gcaggagctg 3600

ggcaagtacg aacagtacat caagtggcca tggtacatct ggctgggatt catcgctggc 3660

ctcatcgcta tcgtgatggt gaccatcatg ctgtgctgca tgacttcatg ctgctcctgc 3720

ctgaagggtt gctgcagctg cggctcttgc tgcaagttcg acgaggacga ctctgaacct 3780

tcctgaaggg agtgaagctg cactacacct aa 3812

<210> 146

<211> 1267

<212> PRT

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 146

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Phe Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Ala

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Gly Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser Gly Trp Thr

245 250 255

Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro Arg Thr Phe

260 265 270

Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala Val Asp Cys

275 280 285

Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser Phe Thr

290 295 300

Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln Pro Thr

305 310 315 320

Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly

325 330 335

Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg

340 345 350

Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser

355 360 365

Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu

370 375 380

Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg

385 390 395 400

Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Asn Ile Ala

405 410 415

Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala

420 425 430

Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr

435 440 445

Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp

450 455 460

Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val

465 470 475 480

Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro

485 490 495

Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe

500 505 510

Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr

515 520 525

Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn Gly Leu Thr

530 535 540

Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro Phe Gln

545 550 555 560

Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val Arg Asp Pro

565 570 575

Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly Val

580 585 590

Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val Ala Val Leu

595 600 605

Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile His Ala Asp

610 615 620

Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser Asn Val Phe

625 630 635 640

Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val Asn Asn Ser

645 650 655

Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser Tyr Gln

660 665 670

Thr Gln Thr Asn Ser Pro Gly Gly Ser Val Ala Ser Gln Ser Ile Ile

675 680 685

Ala Tyr Thr Met Ser Leu Gly Val Glu Asn Ser Val Ala Tyr Ser Asn

690 695 700

Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile Ser Val Thr Thr Glu

705 710 715 720

Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys Thr Met Tyr

725 730 735

Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu Gln Tyr Gly

740 745 750

Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr Gly Ile Ala Val Glu

755 760 765

Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys Gln Ile Tyr

770 775 780

Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Asn Phe Ser Gln Ile Leu

785 790 795 800

Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu

805 810 815

Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly

820 825 830

Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys

835 840 845

Phe Asn Gly Leu Thr Val Leu Pro Pro Leu Leu Thr Asp Glu Met Ile

850 855 860

Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp

865 870 875 880

Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala Met Gln Met

885 890 895

Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val Leu Tyr Glu

900 905 910

Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile

915 920 925

Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp

930 935 940

Val Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu Val Lys Gln Leu

945 950 955 960

Ser Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn Asp Ile Leu Ser

965 970 975

Arg Leu Asp Pro Pro Glu Ala Glu Val Gln Ile Asp Arg Leu Ile Thr

980 985 990

Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg

995 1000 1005

Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala Thr Lys Met

1010 1015 1020

Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val Asp Phe Cys Gly

1025 1030 1035

Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ser Ala Pro His Gly

1040 1045 1050

Val Val Phe Leu His Val Thr Tyr Val Pro Ala Gln Glu Lys Asn

1055 1060 1065

Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys Ala His Phe

1070 1075 1080

Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His Trp Phe Val

1085 1090 1095

Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp Asn

1100 1105 1110

Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn

1115 1120 1125

Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys

1130 1135 1140

Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val

1145 1150 1155

Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile

1160 1165 1170

Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn

1175 1180 1185

Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr

1190 1195 1200

Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile Ala Gly Leu

1205 1210 1215

Ile Ala Ile Val Met Val Thr Ile Met Leu Cys Cys Met Thr Ser

1220 1225 1230

Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly Ser Cys Cys

1235 1240 1245

Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu Lys Gly Val Lys

1250 1255 1260

Leu His Tyr Thr

1265

<210> 147

<211> 1254

<212> PRT

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 147

Gln Cys Val Asn Phe Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Ala Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Gly Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

225 230 235 240

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

245 250 255

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

260 265 270

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

275 280 285

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

290 295 300

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

305 310 315 320

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

325 330 335

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

340 345 350

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

355 360 365

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

370 375 380

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

385 390 395 400

Asn Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

405 410 415

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

420 425 430

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

435 440 445

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

450 455 460

Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

465 470 475 480

Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

485 490 495

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

500 505 510

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

515 520 525

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

530 535 540

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

545 550 555 560

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

565 570 575

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

580 585 590

Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile

595 600 605

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

610 615 620

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

625 630 635 640

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

645 650 655

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gly Gly Ser Val Ala Ser Gln

660 665 670

Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Val Glu Asn Ser Val Ala

675 680 685

Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile Ser Val

690 695 700

Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys

705 710 715 720

Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu

725 730 735

Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr Gly Ile

740 745 750

Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys

755 760 765

Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Asn Phe Ser

770 775 780

Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser Phe Ile Glu

785 790 795 800

Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile Lys

805 810 815

Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile Cys

820 825 830

Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu Leu Thr Asp

835 840 845

Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile Thr

850 855 860

Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe Ala

865 870 875 880

Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn Val

885 890 895

Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala Ile

900 905 910

Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly Lys

915 920 925

Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu Val

930 935 940

Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn Asp

945 950 955 960

Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln Ile Asp Arg

965 970 975

Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln Gln

980 985 990

Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala Ala Thr

995 1000 1005

Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val Asp Phe

1010 1015 1020

Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ser Ala Pro

1025 1030 1035

His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala Gln Glu

1040 1045 1050

Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys Ala

1055 1060 1065

His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His Trp

1070 1075 1080

Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr

1085 1090 1095

Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile

1100 1105 1110

Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser

1115 1120 1125

Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro

1130 1135 1140

Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val

1145 1150 1155

Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn

1160 1165 1170

Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu

1175 1180 1185

Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe Ile Ala

1190 1195 1200

Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu Cys Cys Met

1205 1210 1215

Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys Gly Ser

1220 1225 1230

Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu Lys Gly

1235 1240 1245

Val Lys Leu His Tyr Thr

1250

<210> 148

<211> 3804

<212> DNA

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 148

atgttcgtct tcctggtgct gctgcctctg gtgtccagcc agtgcgtgaa cttcaccact 60

aggactcagc tgcctcccgc ttacaccaac tcattcacta ggggtgtgta ctacccagac 120

aaggtcttcc gctcctcagt gctgcactca actcaggacc tgttcctgcc tttcttctcc 180

aacgtcacct ggttccacgc tatccacgtg tccggcacca acggcactaa gaggttcgcc 240

aaccccgtgc tgccattcaa cgacggagtc tacttcgctt caaccgagaa gtccaacatc 300

atcagaggat ggatcttcgg caccactctg gacagcaaga ctcagtctct gctgatcgtc 360

aacaacgcta ccaacgtggt catcaaggtg tgcgaattcc agttctgtaa cgaccccttc 420

ctgggtgtct actaccacaa gaacaacaag tcatggatgg agtccgaatt ccgcgtctac 480

tccagcgcta acaactgcac tttcgagtac gtgtcccagc cattcctgat ggacctggaa 540

ggaaagcagg gtaacttcaa gaacctgcgc gagttcgtgt tcaagaacat cgacggctac 600

ttcaagattt acagcaagca cacccccatc aacctggtcc gtggcctgcc acagggattc 660

tctgctctgg agcccctggt ggacctgcca atcggcatca acatcaccag gttccagact 720

ctgcacagat cctacctgac tcccggagac tcttcatccg gttggaccgc tggcgctgcc 780

gcttactacg tgggttacct gcagccacgc accttcctgc tgaagtacaa cgaaaacggc 840

accatcactg acgccgtcga ctgcgctctg gacccactgt cagaaaccaa gtgcactctg 900

aagtccttca ccgtggagaa gggaatctac cagacttcaa acttccgcgt gcagcctacc 960

gaatccatcg tccgtttccc aaacatcact aacctgtgcc ctttcggaga ggtcttcaac 1020

gccacccgct tcgcttccgt gtacgcctgg aaccgcaagc gtatctcaaa ctgcgtcgct 1080

gactactccg tgctgtacaa ctcagcctcc ttcagcacct tcaagtgcta cggagtgtca 1140

cccactaagc tgaacgacct gtgcttcacc aacgtctacg ctgactcctt cgtgatccgc 1200

ggtgacgagg tccgtcagat cgctccaggc cagactggaa acatcgccga ctacaactac 1260

aagctgcctg acgacttcac cggttgcgtc atcgcctgga acagcaacaa cctggactct 1320

aaagtgggtg gcaactacaa ctacctgtac aggctgttca gaaagtcaaa cctgaagccc 1380

ttcgagcgcg acatcagcac tgaaatctac caggctggtt ctaccccatg caacggtgtc 1440

aagggcttca actgctactt ccctctgcag tcctacggct tccagcccac ttacggtgtg 1500

ggctaccagc cataccgtgt ggtcgtgctg agcttcgagc tgctgcacgc ccctgctact 1560

gtgtgcggac ccaagaagtc taccaacctg gtcaagaaca agtgcgtgaa cttcaacttc 1620

aacggtctga ccggaactgg tgtcctgacc gagagcaaca agaagttcct gcctttccag 1680

cagttcggtc gcgacatcgc tgacaccact gacgccgtgc gtgaccctca gaccctggaa 1740

atcctggaca tcactccctg ctcattcgga ggtgtctccg tgatcactcc cggaaccaac 1800

acttctaacc aggtcgctgt gctgtaccag ggtgtgaact gcaccgaagt cccagtggcc 1860

atccacgctg accagctgac ccctacttgg agggtgtact ccaccggcag caacgtcttc 1920

cagactagag ctggatgcct gatcggtgcc gagcacgtca acaacagcta cgaatgcgac 1980

atccctatcg gcgctggaat ctgcgcctct taccagaccc agactaacag cccaggtggt 2040

tctgtggcct ctcagtcaat catcgcctac accatgtcac tgggtgtcga aaactccgtg 2100

gcttactcta acaactcaat cgccatccct accaacttca ctatcagcgt gaccactgag 2160

atcctgcccg tcagcatgac caagacttct gtggactgca ctatgtacat ctgcggcgac 2220

agcaccgaat gctctaacct gctgctgcag tacggttctt tctgcaccca gctgaaccgc 2280

gctctgactg gcatcgccgt ggagcaggac aagaacactc aggaagtctt cgctcaggtg 2340

aagcaaatct acaagacccc acctatcaag gacttcggcg gattcaactt ctcccagatc 2400

ctgccagacc cttccaagcc cagcaagagg tctttcatcg aggacctgct gttcaacaag 2460

gtcactctgg ccgacgctgg tttcatcaag cagtacggcg actgcctggg agacatcgct 2520

gctcgcgacc tgatctgcgc ccagaagttc aacggcctga ccgtgctgcc cccactgctg 2580

actgacgaaa tgatcgctca gtacactagc gccctgctgg ctggcaccat cacttcaggt 2640

tggactttcg gtgctggtgc tgctctgcag atcccattcg ctatgcagat ggcctaccgt 2700

ttcaacggaa tcggtgtcac ccagaacgtg ctgtacgaga accagaagct gatcgctaac 2760

cagttcaact cagccatcgg caagatccag gacagcctga gctctactgc ctctgctctg 2820

ggaaagctgc aggacgtcgt gaaccagaac gcccaggctc tgaacaccct ggtcaagcag 2880

ctgtcatcca acttcggcgc catcagctct gtgctgaacg acatcctgtc caggctggac 2940

cctcccgagg ctgaagtcca gatcgacagg ctgatcactg gaagactgca gtcactgcag 3000

acctacgtga ctcagcagct gatccgcgcc gctgaaatcc gtgcctccgc taacctggcc 3060

gctaccaaga tgagcgagtg cgtcctgggc cagtctaaga gagtggactt ctgcggcaag 3120

ggataccacc tgatgtcatt cccacagtcc gcccctcacg gagtcgtgtt cctgcacgtc 3180

acctacgtgc ccgctcagga gaagaacttc accactgccc cagctatctg ccacgacggc 3240

aaggctcact tccctcgcga aggagtcttc gtgagcaacg gcacccactg gttcgtcact 3300

cagcgtaact tctacgagcc ccagatcatc accactgaca acactttcgt gtctggcaac 3360

tgcgacgtgg tcatcggtat cgtcaacaac accgtgtacg accctctgca gcccgagctg 3420

gactcattca aggaagagct ggacaagtac ttcaagaacc acacctcccc tgacgtcgac 3480

ctgggtgaca tctcaggcat caacgcctcc gtcgtgaaca tccagaagga gatcgaccgc 3540

ctgaacgaag tggctaagaa cctgaacgaa agcctgatcg acctgcagga gctgggcaag 3600

tacgaacagt acatcaagtg gccatggtac atctggctgg gattcatcgc tggcctcatc 3660

gctatcgtga tggtgaccat catgctgtgc tgcatgactt catgctgctc ctgcctgaag 3720

ggttgctgca gctgcggctc ttgctgcaag ttcgacgagg acgactctga acctgtcctg 3780

aagggagtga agctgcacta cacc 3804

<210> 149

<211> 1273

<212> PRT

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 149

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Phe Thr Asn Arg Thr Gln Leu Pro Ser Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Tyr Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Ser Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Thr Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu Tyr Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln Ser Val Ala

675 680 685

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser

690 695 700

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

705 710 715 720

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

725 730 735

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

740 745 750

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

755 760 765

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

770 775 780

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe

785 790 795 800

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

805 810 815

Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly

820 825 830

Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp

835 840 845

Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu

850 855 860

Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly

865 870 875 880

Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile

885 890 895

Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr

900 905 910

Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn

915 920 925

Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala

930 935 940

Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn

945 950 955 960

Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val

965 970 975

Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln

980 985 990

Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val

995 1000 1005

Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn

1010 1015 1020

Leu Ala Ala Ile Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

1025 1030 1035

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro

1040 1045 1050

Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val

1055 1060 1065

Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

1070 1075 1080

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

1085 1090 1095

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln

1100 1105 1110

Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val

1115 1120 1125

Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro

1130 1135 1140

Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn

1145 1150 1155

His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn

1160 1165 1170

Ala Ser Phe Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu

1175 1180 1185

Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu

1190 1195 1200

Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu

1205 1210 1215

Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met

1220 1225 1230

Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys

1235 1240 1245

Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro

1250 1255 1260

Val Leu Lys Gly Val Lys Leu His Tyr Thr

1265 1270

<210> 150

<211> 3822

<212> DNA

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 150

atgttcgtct tcctggtgct gctgcctctg gtgtccagcc agtgcgtgaa cttcaccaac 60

cgcactcagc tgcctagcgc ttacaccaac tctttcacta ggggtgtgta ctaccccgac 120

aaggtcttcc gctcctcagt gctgcactct actcaggacc tgttcctgcc cttcttctct 180

aacgtcacct ggttccacgc catccacgtg agcggcacca acggcactaa gaggttcgac 240

aacccagtgc tgcctttcaa cgacggtgtc tacttcgctt caaccgagaa gtccaacatc 300

atcagaggat ggatcttcgg caccactctg gacagcaaga ctcagtctct gctgatcgtc 360

aacaacgcca ccaacgtggt catcaaggtg tgcgaattcc agttctgcaa ctacccattc 420

ctgggcgtct actaccacaa gaacaacaag tcatggatgg agtccgaatt ccgcgtctac 480

tccagcgcta acaactgcac tttcgagtac gtgtcccagc ctttcctgat ggacctggaa 540

ggaaagcagg gtaacttcaa gaacctgtca gagttcgtgt tcaagaacat cgacggatac 600

ttcaagattt actccaagca caccccaatc aacctggtcc gtgacctgcc tcagggtttc 660

tcagctctgg agccactggt ggacctgcct atcggcatca acatcaccag gttccagact 720

ctgctggctc tgcacagatc ctacctgact ccaggtgact catcttctgg atggactgct 780

ggagctgctg cttactacgt gggttacctg cagcctcgca ccttcctgct gaagtacaac 840

gaaaacggca ccatcactga cgccgtcgac tgcgctctgg accctctgag cgaaaccaag 900

tgcactctga agtctttcac cgtggagaag ggtatctacc agactagcaa cttccgcgtg 960

cagcccaccg aatctatcgt ccgtttccct aacatcacta acctgtgccc cttcggcgag 1020

gtcttcaacg ccacccgttt cgcttccgtg tacgcctgga accgcaagcg tatcagcaac 1080

tgcgtcgctg actactctgt gctgtacaac tcagcctcct tcagcacttt caagtgctac 1140

ggtgtgagcc ctactaagct gaacgacctg tgcttcacca acgtctacgc cgactctttc 1200

gtgatccgcg gcgacgaggt ccgtcagatc gctccaggcc agaccggaac tatcgccgac 1260

tacaactaca agctgcctga cgacttcacc ggctgcgtca tcgcttggaa ctcaaacaac 1320

ctggactcca aagtgggtgg caactacaac tacctgtaca ggctgttcag aaagtcaaac 1380

ctgaagccct tcgagaggga catctcaact gaaatctacc aggccggctc caccccatgc 1440

aacggtgtca agggcttcaa ctgctacttc cccctgcagt cctacggatt ccagccaact 1500

tacggtgtgg gctaccagcc ttacagagtg gtcgtgctgt cattcgaact cctccacgct 1560

cctgctactg tgtgcggacc aaagaagtcc accaacctgg tcaagaacaa gtgcgtgaac 1620

ttcaacttca acggtctgac cggaactggt gtcctgaccg agtcaaacaa gaagttcctg 1680

cccttccagc agttcggtcg cgacatcgct gacactaccg acgccgtgcg tgacccccag 1740

accctggaaa tcctggacat cactccatgc agcttcggag gtgtctctgt gatcactcca 1800

ggaaccaaca cttccaacca ggtcgctgtg ctgtaccagg gtgtcaactg caccgaagtc 1860

cctgtggcca tccacgctga ccagctgacc cccacttgga gggtgtactc caccggcagc 1920

aacgtcttcc agactagagc tggttgcctg atcggcgccg agtacgtgaa caactcatac 1980

gaatgcgaca tccctatcgg cgctggaatc tgcgcctcct accagaccca gactaactca 2040

ccccagcagg ctcagtccgt cgcctctcag tcaatcatcg cttacaccat gagcctgggc 2100

gctgaaaact ctgtggccta ctctaacaac tcaatcgcca tccccaccaa cttcactatc 2160

agcgtgacca ctgagatcct gccagtctca atgaccaaga cttccgtgga ctgcactatg 2220

tacatctgcg gagacagcac cgaatgctct aacctgctgc tgcagtacgg ctctttctgc 2280

acccagctga acagggctct gactggaatc gccgtggagc aggacaagaa cactcaggaa 2340

gtcttcgctc aggtgaagca aatctacaag acccctccca tcaaggactt cggcggattc 2400

aacttctccc agatcctgcc tgacccctcc aagcccagca agcgctcttt catcgaggac 2460

ctgctgttca acaaggtcac cctggccgac gctggcttca tcaagcagta cggagactgc 2520

ctgggtgaca tcgccgctcg tgacctgatc tgcgctcaga agttcaacgg actgaccgtg 2580

ctgccacctc tgctgactga cgaaatgatc gcccagtaca cttcagccct gctggctgga 2640

accatcactt ctggttggac cttcggtgct ggtgctgctc tgcagatccc attcgctatg 2700

cagatggcct accgcttcaa cggaatcggt gtcacccaga acgtgctgta cgagaaccag 2760

aagctgatcg ctaaccagtt caacagcgcc atcggcaaga tccaggactc actgagctct 2820

actgcctccg ctctgggaaa gctgcaggac gtcgtgaacc agaacgccca ggctctgaac 2880

accctggtca agcagctgtc atccaacttc ggagctatca gctctgtgct gaacgacatc 2940

ctctcccgcc tggaccctcc tgaggctgaa gtccagatcg acaggctgat cactggtaga 3000

ctgcagagcc tgcagaccta cgtgactcag cagctgatcc gcgccgctga aatccgtgcc 3060

tctgctaacc tggctgctat caagatgtca gagtgcgtcc tgggacagtc caagagagtg 3120

gacttctgcg gcaagggata ccacctgatg tcattccctc agtccgctcc ccacggtgtc 3180

gtgttcctgc acgtcaccta cgtgccagcc caggagaaga acttcaccac tgcccctgct 3240

atctgccacg acggcaaggc tcacttcccc cgcgaaggtg tcttcgtgag caacggcacc 3300

cactggttcg tcactcagcg taacttctac gagccacaga tcatcaccac tgacaacact 3360

ttcgtgtctg gaaactgcga cgtggtcatc ggtatcgtca acaacaccgt gtacgacccc 3420

ctgcagccag agctggacag cttcaaggaa gagctggaca agtacttcaa gaaccacacc 3480

tctcccgacg tggacctggg cgacatcagc ggaatcaacg cttctttcgt caacatccag 3540

aaggagatcg accgtctgaa cgaagtggcc aagaacctga acgaatcact gatcgacctg 3600

caggagctgg gcaagtacga acagtacatc aagtggccat ggtacatctg gctgggtttc 3660

atcgctggcc tcatcgctat cgtgatggtg accatcatgc tgtgctgcat gacttcatgc 3720

tgctcctgcc tgaagggctg ctgcagctgc ggatcttgct gcaagttcga cgaggacgac 3780

tccgaacctg tcctgaaggg agtgaagctg cactacacct aa 3822

<210> 151

<211> 1260

<212> PRT

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 151

Gln Cys Val Asn Phe Thr Asn Arg Thr Gln Leu Pro Ser Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Tyr Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Ser Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Thr Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu Tyr Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln

660 665 670

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala

675 680 685

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn

690 695 700

Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys

705 710 715 720

Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys

725 730 735

Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg

740 745 750

Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val

755 760 765

Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe

770 775 780

Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser

785 790 795 800

Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala

805 810 815

Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala

820 825 830

Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu

835 840 845

Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu

850 855 860

Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala

865 870 875 880

Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile

885 890 895

Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn

900 905 910

Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr

915 920 925

Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln

930 935 940

Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile

945 950 955 960

Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala

965 970 975

Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln

980 985 990

Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser

995 1000 1005

Ala Asn Leu Ala Ala Ile Lys Met Ser Glu Cys Val Leu Gly Gln

1010 1015 1020

Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser

1025 1030 1035

Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr

1040 1045 1050

Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile

1055 1060 1065

Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val

1070 1075 1080

Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu

1085 1090 1095

Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys

1100 1105 1110

Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu

1115 1120 1125

Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe

1130 1135 1140

Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly

1145 1150 1155

Ile Asn Ala Ser Phe Val Asn Ile Gln Lys Glu Ile Asp Arg Leu

1160 1165 1170

Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln

1175 1180 1185

Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile

1190 1195 1200

Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr

1205 1210 1215

Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly

1220 1225 1230

Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser

1235 1240 1245

Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255 1260

<210> 152

<211> 1273

<212> PRT

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 152

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ile Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Cys Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Arg Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Pro Tyr Gly

485 490 495

Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln Ser Val Ala

675 680 685

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser

690 695 700

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

705 710 715 720

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

725 730 735

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

740 745 750

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

755 760 765

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

770 775 780

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe

785 790 795 800

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

805 810 815

Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly

820 825 830

Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp

835 840 845

Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu

850 855 860

Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly

865 870 875 880

Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile

885 890 895

Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr

900 905 910

Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn

915 920 925

Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala

930 935 940

Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn

945 950 955 960

Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val

965 970 975

Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln

980 985 990

Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val

995 1000 1005

Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn

1010 1015 1020

Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

1025 1030 1035

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro

1040 1045 1050

Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val

1055 1060 1065

Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

1070 1075 1080

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

1085 1090 1095

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln

1100 1105 1110

Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val

1115 1120 1125

Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro

1130 1135 1140

Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn

1145 1150 1155

His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn

1160 1165 1170

Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu

1175 1180 1185

Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu

1190 1195 1200

Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu

1205 1210 1215

Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met

1220 1225 1230

Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys

1235 1240 1245

Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro

1250 1255 1260

Val Leu Lys Gly Val Lys Leu His Tyr Thr

1265 1270

<210> 153

<211> 1260

<212> PRT

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 153

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Cys Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Arg Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Pro Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln

660 665 670

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala

675 680 685

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn

690 695 700

Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys

705 710 715 720

Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys

725 730 735

Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg

740 745 750

Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val

755 760 765

Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe

770 775 780

Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser

785 790 795 800

Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala

805 810 815

Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala

820 825 830

Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu

835 840 845

Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu

850 855 860

Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala

865 870 875 880

Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile

885 890 895

Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn

900 905 910

Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr

915 920 925

Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln

930 935 940

Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile

945 950 955 960

Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala

965 970 975

Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln

980 985 990

Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser

995 1000 1005

Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln

1010 1015 1020

Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser

1025 1030 1035

Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr

1040 1045 1050

Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile

1055 1060 1065

Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val

1070 1075 1080

Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu

1085 1090 1095

Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys

1100 1105 1110

Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu

1115 1120 1125

Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe

1130 1135 1140

Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly

1145 1150 1155

Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu

1160 1165 1170

Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln

1175 1180 1185

Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile

1190 1195 1200

Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr

1205 1210 1215

Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly

1220 1225 1230

Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser

1235 1240 1245

Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255 1260

<210> 154

<211> 13

<212> PRT

<213> 人工序列

<220>

<223> N-末端信号肽

<400> 154

Met Phe Val Phe Phe Val Leu Leu Pro Leu Val Ser Ser

1 5 10

<210> 155

<211> 1273

<212> PRT

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 155

Met Phe Val Phe Phe Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Ile Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Gly Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln Ser Val Ala

675 680 685

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Val Glu Asn Ser

690 695 700

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

705 710 715 720

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

725 730 735

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

740 745 750

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

755 760 765

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

770 775 780

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe

785 790 795 800

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

805 810 815

Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly

820 825 830

Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp

835 840 845

Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu

850 855 860

Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly

865 870 875 880

Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile

885 890 895

Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr

900 905 910

Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn

915 920 925

Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala

930 935 940

Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn

945 950 955 960

Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val

965 970 975

Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln

980 985 990

Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val

995 1000 1005

Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn

1010 1015 1020

Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

1025 1030 1035

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro

1040 1045 1050

Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val

1055 1060 1065

Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

1070 1075 1080

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

1085 1090 1095

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln

1100 1105 1110

Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val

1115 1120 1125

Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro

1130 1135 1140

Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn

1145 1150 1155

His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn

1160 1165 1170

Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu

1175 1180 1185

Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu

1190 1195 1200

Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu

1205 1210 1215

Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met

1220 1225 1230

Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys

1235 1240 1245

Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro

1250 1255 1260

Val Leu Lys Gly Val Lys Leu His Tyr Thr

1265 1270

<210> 156

<211> 1260

<212> PRT

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 156

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Ile Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Gly

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln

660 665 670

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Val

675 680 685

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn

690 695 700

Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys

705 710 715 720

Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys

725 730 735

Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg

740 745 750

Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val

755 760 765

Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe

770 775 780

Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser

785 790 795 800

Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala

805 810 815

Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala

820 825 830

Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu

835 840 845

Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu

850 855 860

Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala

865 870 875 880

Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile

885 890 895

Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn

900 905 910

Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr

915 920 925

Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln

930 935 940

Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile

945 950 955 960

Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala

965 970 975

Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln

980 985 990

Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser

995 1000 1005

Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln

1010 1015 1020

Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser

1025 1030 1035

Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr

1040 1045 1050

Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile

1055 1060 1065

Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val

1070 1075 1080

Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu

1085 1090 1095

Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys

1100 1105 1110

Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu

1115 1120 1125

Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe

1130 1135 1140

Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly

1145 1150 1155

Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu

1160 1165 1170

Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln

1175 1180 1185

Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile

1190 1195 1200

Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr

1205 1210 1215

Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly

1220 1225 1230

Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser

1235 1240 1245

Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255 1260

<210> 157

<211> 1273

<212> PRT

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 157

Met Phe Val Phe Phe Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Ile Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Gly Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Asn Thr Pro Cys

465 470 475 480

Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln Ser Val Ala

675 680 685

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Val Glu Asn Ser

690 695 700

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

705 710 715 720

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

725 730 735

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

740 745 750

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

755 760 765

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

770 775 780

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe

785 790 795 800

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

805 810 815

Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly

820 825 830

Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp

835 840 845

Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu

850 855 860

Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly

865 870 875 880

Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile

885 890 895

Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr

900 905 910

Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn

915 920 925

Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala

930 935 940

Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn

945 950 955 960

Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val

965 970 975

Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln

980 985 990

Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val

995 1000 1005

Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn

1010 1015 1020

Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

1025 1030 1035

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro

1040 1045 1050

Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val

1055 1060 1065

Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

1070 1075 1080

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

1085 1090 1095

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln

1100 1105 1110

Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val

1115 1120 1125

Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro

1130 1135 1140

Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn

1145 1150 1155

His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn

1160 1165 1170

Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu

1175 1180 1185

Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu

1190 1195 1200

Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu

1205 1210 1215

Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met

1220 1225 1230

Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys

1235 1240 1245

Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro

1250 1255 1260

Val Leu Lys Gly Val Lys Leu His Tyr Thr

1265 1270

<210> 158

<211> 1260

<212> PRT

<213> 人工序列

<220>

<223> 修饰的冠状病毒S多肽

<400> 158

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Ile Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Gly

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Asn

450 455 460

Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Gln Gln Ala Gln

660 665 670

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Val

675 680 685

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn

690 695 700

Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys

705 710 715 720

Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys

725 730 735

Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg

740 745 750

Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val

755 760 765

Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe

770 775 780

Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser

785 790 795 800

Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala

805 810 815

Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala

820 825 830

Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu

835 840 845

Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu

850 855 860

Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala

865 870 875 880

Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile

885 890 895

Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn

900 905 910

Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr

915 920 925

Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln

930 935 940

Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile

945 950 955 960

Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala

965 970 975

Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln

980 985 990

Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser

995 1000 1005

Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln

1010 1015 1020

Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser

1025 1030 1035

Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr

1040 1045 1050

Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile

1055 1060 1065

Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val

1070 1075 1080

Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu

1085 1090 1095

Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys

1100 1105 1110

Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu

1115 1120 1125

Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe

1130 1135 1140

Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly

1145 1150 1155

Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu

1160 1165 1170

Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln

1175 1180 1185

Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile

1190 1195 1200

Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr

1205 1210 1215

Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly

1220 1225 1230

Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser

1235 1240 1245

Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255 1260

<210> 159

<211> 1271

<212> PRT

<213> 人工序列

<220>

<223> 重组SARS-CoV-2 S多肽

<400> 159

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Arg Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Asp Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Gly Val Tyr Ser Ser

145 150 155 160

Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu Met Asp

165 170 175

Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe Val Phe

180 185 190

Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr Pro Ile

195 200 205

Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu Pro Leu

210 215 220

Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr Leu Leu

225 230 235 240

Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser Gly Trp

245 250 255

Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro Arg Thr

260 265 270

Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala Val Asp

275 280 285

Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser Phe

290 295 300

Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln Pro

305 310 315 320

Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe

325 330 335

Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn

340 345 350

Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn

355 360 365

Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys

370 375 380

Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile

385 390 395 400

Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile

405 410 415

Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile

420 425 430

Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn

435 440 445

Tyr Arg Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg

450 455 460

Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Lys Pro Cys Asn Gly

465 470 475 480

Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln

485 490 495

Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser

500 505 510

Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser

515 520 525

Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn Gly Leu

530 535 540

Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro Phe

545 550 555 560

Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val Arg Asp

565 570 575

Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly

580 585 590

Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val Ala Val

595 600 605

Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile His Ala

610 615 620

Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser Asn Val

625 630 635 640

Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val Asn Asn

645 650 655

Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser Tyr

660 665 670

Gln Thr Gln Thr Asn Ser Arg Gln Gln Ala Gln Ser Val Ala Ser Gln

675 680 685

Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser Val Ala

690 695 700

Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile Ser Val

705 710 715 720

Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys

725 730 735

Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu

740 745 750

Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr Gly Ile

755 760 765

Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys

770 775 780

Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe Asn Phe

785 790 795 800

Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser Phe Ile

805 810 815

Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile

820 825 830

Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile

835 840 845

Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu Leu Thr

850 855 860

Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile

865 870 875 880

Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe

885 890 895

Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn

900 905 910

Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala

915 920 925

Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly

930 935 940

Lys Leu Gln Asn Val Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu

945 950 955 960

Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn

965 970 975

Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln Ile Asp

980 985 990

Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln

995 1000 1005

Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala

1010 1015 1020

Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val

1025 1030 1035

Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ser

1040 1045 1050

Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala

1055 1060 1065

Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly

1070 1075 1080

Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr

1085 1090 1095

His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile

1100 1105 1110

Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile

1115 1120 1125

Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu

1130 1135 1140

Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr

1145 1150 1155

Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser

1160 1165 1170

Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala

1175 1180 1185

Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys

1190 1195 1200

Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe

1205 1210 1215

Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu Cys

1220 1225 1230

Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys

1235 1240 1245

Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu

1250 1255 1260

Lys Gly Val Lys Leu His Tyr Thr

1265 1270

<210> 160

<211> 1273

<212> PRT

<213> 人工序列

<220>

<223> 重组SARS-CoV-2 S多肽

<400> 160

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Ile Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Asp Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Lys Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Arg Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Gln Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser Arg Gln Gln Ala Gln Ser Val Ala

675 680 685

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser

690 695 700

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

705 710 715 720

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

725 730 735

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

740 745 750

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

755 760 765

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

770 775 780

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe

785 790 795 800

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

805 810 815

Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly

820 825 830

Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp

835 840 845

Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu

850 855 860

Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly

865 870 875 880

Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile

885 890 895

Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr

900 905 910

Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn

915 920 925

Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala

930 935 940

Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn

945 950 955 960

Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val

965 970 975

Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln

980 985 990

Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val

995 1000 1005

Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn

1010 1015 1020

Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

1025 1030 1035

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro

1040 1045 1050

Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val

1055 1060 1065

Pro Ala His Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

1070 1075 1080

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

1085 1090 1095

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln

1100 1105 1110

Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val

1115 1120 1125

Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro

1130 1135 1140

Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn

1145 1150 1155

His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn

1160 1165 1170

Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu

1175 1180 1185

Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu

1190 1195 1200

Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu

1205 1210 1215

Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met

1220 1225 1230

Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys

1235 1240 1245

Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro

1250 1255 1260

Val Leu Lys Gly Val Lys Leu His Tyr Thr

1265 1270

<210> 161

<211> 1271

<212> PRT

<213> 人工序列

<220>

<223> 重组SARS-CoV-2 S多肽

<400> 161

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Arg Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Asp Val Tyr

130 135 140

Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Gly Val Tyr Ser Ser

145 150 155 160

Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu Met Asp

165 170 175

Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe Val Phe

180 185 190

Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr Pro Ile

195 200 205

Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu Pro Leu

210 215 220

Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr Leu Leu

225 230 235 240

Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser Gly Ile

245 250 255

Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro Arg Thr

260 265 270

Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala Val Asp

275 280 285

Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys Ser Phe

290 295 300

Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val Gln Pro

305 310 315 320

Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe

325 330 335

Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn

340 345 350

Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn

355 360 365

Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys

370 375 380

Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile

385 390 395 400

Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Asn Ile

405 410 415

Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile

420 425 430

Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn

435 440 445

Tyr Arg Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg

450 455 460

Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Lys Pro Cys Asn Gly

465 470 475 480

Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln

485 490 495

Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser

500 505 510

Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser

515 520 525

Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn Gly Leu

530 535 540

Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu Pro Phe

545 550 555 560

Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val Arg Asp

565 570 575

Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe Gly Gly

580 585 590

Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val Ala Val

595 600 605

Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile His Ala

610 615 620

Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser Asn Val

625 630 635 640

Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val Asn Asn

645 650 655

Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala Ser Tyr

660 665 670

Gln Thr Gln Thr Asn Ser Arg Gln Gln Ala Gln Ser Val Ala Ser Gln

675 680 685

Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser Val Ala

690 695 700

Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile Ser Val

705 710 715 720

Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val Asp Cys

725 730 735

Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu Leu Leu

740 745 750

Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr Gly Ile

755 760 765

Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln Val Lys

770 775 780

Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe Asn Phe

785 790 795 800

Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser Phe Ile

805 810 815

Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Ile

820 825 830

Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp Leu Ile

835 840 845

Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu Leu Thr

850 855 860

Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly Thr Ile

865 870 875 880

Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile Pro Phe

885 890 895

Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn

900 905 910

Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn Ser Ala

915 920 925

Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala Leu Gly

930 935 940

Lys Leu Gln Asn Val Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu

945 950 955 960

Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val Leu Asn

965 970 975

Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln Ile Asp

980 985 990

Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val Thr Gln

995 1000 1005

Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu Ala

1010 1015 1020

Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg Val

1025 1030 1035

Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ser

1040 1045 1050

Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala

1055 1060 1065

Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly

1070 1075 1080

Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr

1085 1090 1095

His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile

1100 1105 1110

Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile

1115 1120 1125

Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu

1130 1135 1140

Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr

1145 1150 1155

Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser

1160 1165 1170

Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala

1175 1180 1185

Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys

1190 1195 1200

Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly Phe

1205 1210 1215

Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu Cys

1220 1225 1230

Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser Cys

1235 1240 1245

Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val Leu

1250 1255 1260

Lys Gly Val Lys Leu His Tyr Thr

1265 1270

<210> 162

<211> 1273

<212> PRT

<213> 人工序列

<220>

<223> 重组SARS-CoV-2 S多肽

<400> 162

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Ile Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Ser

130 135 140

Asn His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

145 150 155 160

Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

165 170 175

Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

210 215 220

Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

225 230 235 240

Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

275 280 285

Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

325 330 335

Pro Phe Gly Glu Val Phe Asn Ala Thr Lys Phe Ala Ser Val Tyr Ala

340 345 350

Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

370 375 380

Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

405 410 415

Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

420 425 430

Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

435 440 445

Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

450 455 460

Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

465 470 475 480

Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

485 490 495

Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

500 505 510

Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

515 520 525

Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

530 535 540

Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

545 550 555 560

Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

565 570 575

Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

580 585 590

Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

595 600 605

Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala Ile

610 615 620

His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

625 630 635 640

Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

645 650 655

Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

660 665 670

Ser Tyr Gln Thr Gln Thr Asn Ser His Gln Gln Ala Gln Ser Val Ala

675 680 685

Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser

690 695 700

Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

705 710 715 720

Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

725 730 735

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

740 745 750

Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

755 760 765

Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

770 775 780

Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe

785 790 795 800

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

805 810 815

Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly

820 825 830

Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp

835 840 845

Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu

850 855 860

Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly

865 870 875 880

Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile

885 890 895

Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr

900 905 910

Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn

915 920 925

Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala

930 935 940

Leu Gly Lys Leu Gln Asn Val Val Asn Gln Asn Ala Gln Ala Leu Asn

945 950 955 960

Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val

965 970 975

Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val Gln

980 985 990

Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val

995 1000 1005

Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn

1010 1015 1020

Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

1025 1030 1035

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro

1040 1045 1050

Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val

1055 1060 1065

Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

1070 1075 1080

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

1085 1090 1095

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln

1100 1105 1110

Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val

1115 1120 1125

Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro

1130 1135 1140

Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn

1145 1150 1155

His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn

1160 1165 1170

Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu

1175 1180 1185

Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu

1190 1195 1200

Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu

1205 1210 1215

Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met

1220 1225 1230

Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys

1235 1240 1245

Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro

1250 1255 1260

Val Leu Lys Gly Val Lys Leu His Tyr Thr

1265 1270

<210> 163

<211> 1274

<212> PRT

<213> 人工序列

<220>

<223> 重组SARS-CoV-2 S多肽

<400> 163

Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

1 5 10 15

Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

20 25 30

Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

35 40 45

His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

50 55 60

Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

65 70 75 80

Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Ile Glu

85 90 95

Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

100 105 110

Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

115 120 125

Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Thr

130 135 140

Ser Asn His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val

145 150 155 160

Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe

165 170 175

Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu

180 185 190

Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His

195 200 205

Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu

210 215 220

Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln

225 230 235 240

Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser

245 250 255

Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln

260 265 270

Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp

275 280 285

Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu

290 295 300

Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg

305 310 315 320

Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu

325 330 335

Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Lys Phe Ala Ser Val Tyr

340 345 350

Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val

355 360 365

Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser

370 375 380

Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser

385 390 395 400

Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr

405 410 415

Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly

420 425 430

Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly

435 440 445

Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro

450 455 460

Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro

465 470 475 480

Cys Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr

485 490 495

Gly Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val Val

500 505 510

Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro

515 520 525

Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe

530 535 540

Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe

545 550 555 560

Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala

565 570 575

Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser

580 585 590

Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln

595 600 605

Val Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala

610 615 620

Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly

625 630 635 640

Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His

645 650 655

Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys

660 665 670

Ala Ser Tyr Gln Thr Gln Thr Asn Ser His Gln Gln Ala Gln Ser Val

675 680 685

Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn

690 695 700

Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr

705 710 715 720

Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser

725 730 735

Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn

740 745 750

Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu

755 760 765

Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala

770 775 780

Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly

785 790 795 800

Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg

805 810 815

Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala

820 825 830

Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg

835 840 845

Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro

850 855 860

Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala

865 870 875 880

Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln

885 890 895

Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val

900 905 910

Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe

915 920 925

Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser

930 935 940

Ala Leu Gly Lys Leu Gln Asn Val Val Asn Gln Asn Ala Gln Ala Leu

945 950 955 960

Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser

965 970 975

Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val

980 985 990

Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr

995 1000 1005

Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala

1010 1015 1020

Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser

1025 1030 1035

Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe

1040 1045 1050

Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr

1055 1060 1065

Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys

1070 1075 1080

His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser

1085 1090 1095

Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro

1100 1105 1110

Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp

1115 1120 1125

Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln

1130 1135 1140

Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys

1145 1150 1155

Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile

1160 1165 1170

Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn

1175 1180 1185

Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu

1190 1195 1200

Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp

1205 1210 1215

Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile

1220 1225 1230

Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys

1235 1240 1245

Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu

1250 1255 1260

Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

1265 1270

<210> 164

<211> 1258

<212> PRT

<213> 人工序列

<220>

<223> 重组SARS-CoV-2 S多肽

<400> 164

Gln Cys Val Asn Leu Arg Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Asp Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Gly Val

130 135 140

Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe

145 150 155 160

Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu

165 170 175

Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His

180 185 190

Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu

195 200 205

Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln

210 215 220

Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser

225 230 235 240

Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln

245 250 255

Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp

260 265 270

Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu

275 280 285

Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg

290 295 300

Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu

305 310 315 320

Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr

325 330 335

Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val

340 345 350

Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser

355 360 365

Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser

370 375 380

Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr

385 390 395 400

Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly

405 410 415

Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly

420 425 430

Asn Tyr Asn Tyr Arg Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro

435 440 445

Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Lys Pro

450 455 460

Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr

465 470 475 480

Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val

485 490 495

Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro

500 505 510

Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe

515 520 525

Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe

530 535 540

Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala

545 550 555 560

Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser

565 570 575

Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln

580 585 590

Val Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala

595 600 605

Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly

610 615 620

Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His

625 630 635 640

Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys

645 650 655

Ala Ser Tyr Gln Thr Gln Thr Asn Ser Arg Gln Gln Ala Gln Ser Val

660 665 670

Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn

675 680 685

Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr

690 695 700

Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser

705 710 715 720

Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn

725 730 735

Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu

740 745 750

Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala

755 760 765

Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly

770 775 780

Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg

785 790 795 800

Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala

805 810 815

Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg

820 825 830

Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro

835 840 845

Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala

850 855 860

Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln

865 870 875 880

Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val

885 890 895

Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe

900 905 910

Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser

915 920 925

Ala Leu Gly Lys Leu Gln Asn Val Val Asn Gln Asn Ala Gln Ala Leu

930 935 940

Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser

945 950 955 960

Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val

965 970 975

Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr

980 985 990

Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn

995 1000 1005

Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

1010 1015 1020

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro

1025 1030 1035

Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val

1040 1045 1050

Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

1055 1060 1065

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

1070 1075 1080

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln

1085 1090 1095

Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val

1100 1105 1110

Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro

1115 1120 1125

Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn

1130 1135 1140

His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn

1145 1150 1155

Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu

1160 1165 1170

Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu

1175 1180 1185

Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu

1190 1195 1200

Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met

1205 1210 1215

Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys

1220 1225 1230

Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro

1235 1240 1245

Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255

<210> 165

<211> 1260

<212> PRT

<213> 人工序列

<220>

<223> 重组SARS-CoV-2 S多肽

<400> 165

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Ile Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Asp Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Lys Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Arg Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Gln Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Arg Gln Gln Ala Gln

660 665 670

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala

675 680 685

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn

690 695 700

Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys

705 710 715 720

Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys

725 730 735

Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg

740 745 750

Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val

755 760 765

Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe

770 775 780

Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser

785 790 795 800

Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala

805 810 815

Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala

820 825 830

Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu

835 840 845

Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu

850 855 860

Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala

865 870 875 880

Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile

885 890 895

Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn

900 905 910

Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr

915 920 925

Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln

930 935 940

Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile

945 950 955 960

Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala

965 970 975

Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln

980 985 990

Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser

995 1000 1005

Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln

1010 1015 1020

Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser

1025 1030 1035

Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr

1040 1045 1050

Tyr Val Pro Ala His Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile

1055 1060 1065

Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val

1070 1075 1080

Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu

1085 1090 1095

Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys

1100 1105 1110

Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu

1115 1120 1125

Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe

1130 1135 1140

Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly

1145 1150 1155

Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu

1160 1165 1170

Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln

1175 1180 1185

Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile

1190 1195 1200

Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr

1205 1210 1215

Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly

1220 1225 1230

Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser

1235 1240 1245

Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255 1260

<210> 166

<211> 1258

<212> PRT

<213> 人工序列

<220>

<223> 重组SARS-CoV-2 S多肽

<400> 166

Gln Cys Val Asn Leu Arg Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Asp Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Gly Val

130 135 140

Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe

145 150 155 160

Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu

165 170 175

Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His

180 185 190

Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu

195 200 205

Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln

210 215 220

Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser

225 230 235 240

Ser Gly Ile Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln

245 250 255

Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp

260 265 270

Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu

275 280 285

Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg

290 295 300

Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu

305 310 315 320

Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr

325 330 335

Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val

340 345 350

Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser

355 360 365

Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser

370 375 380

Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr

385 390 395 400

Gly Asn Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly

405 410 415

Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly

420 425 430

Asn Tyr Asn Tyr Arg Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro

435 440 445

Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Lys Pro

450 455 460

Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr

465 470 475 480

Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val

485 490 495

Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro

500 505 510

Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe

515 520 525

Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe

530 535 540

Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala

545 550 555 560

Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser

565 570 575

Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln

580 585 590

Val Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro Val Ala

595 600 605

Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly

610 615 620

Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His

625 630 635 640

Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys

645 650 655

Ala Ser Tyr Gln Thr Gln Thr Asn Ser Arg Gln Gln Ala Gln Ser Val

660 665 670

Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn

675 680 685

Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr

690 695 700

Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser

705 710 715 720

Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn

725 730 735

Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu

740 745 750

Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala

755 760 765

Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly

770 775 780

Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg

785 790 795 800

Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala

805 810 815

Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg

820 825 830

Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro

835 840 845

Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala

850 855 860

Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln

865 870 875 880

Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val

885 890 895

Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe

900 905 910

Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser

915 920 925

Ala Leu Gly Lys Leu Gln Asn Val Val Asn Gln Asn Ala Gln Ala Leu

930 935 940

Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser

945 950 955 960

Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala Glu Val

965 970 975

Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr

980 985 990

Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn

995 1000 1005

Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

1010 1015 1020

Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro

1025 1030 1035

Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val

1040 1045 1050

Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

1055 1060 1065

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

1070 1075 1080

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln

1085 1090 1095

Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val

1100 1105 1110

Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro

1115 1120 1125

Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn

1130 1135 1140

His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn

1145 1150 1155

Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu

1160 1165 1170

Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu

1175 1180 1185

Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu

1190 1195 1200

Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met

1205 1210 1215

Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys

1220 1225 1230

Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro

1235 1240 1245

Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255

<210> 167

<211> 1260

<212> PRT

<213> 人工序列

<220>

<223> 重组SARS-CoV-2 S多肽

<400> 167

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Ile Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Ser Asn His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe

130 135 140

Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln

145 150 155 160

Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu

165 170 175

Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser

180 185 190

Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser

195 200 205

Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg

210 215 220

Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp

225 230 235 240

Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr

245 250 255

Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile

260 265 270

Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys

275 280 285

Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn

290 295 300

Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr

305 310 315 320

Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Lys Phe Ala Ser

325 330 335

Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr

340 345 350

Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly

355 360 365

Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala

370 375 380

Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly

385 390 395 400

Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe

405 410 415

Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val

420 425 430

Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu

435 440 445

Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser

450 455 460

Thr Pro Cys Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln

465 470 475 480

Ser Tyr Gly Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg

485 490 495

Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys

500 505 510

Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe

515 520 525

Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys

530 535 540

Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr

545 550 555 560

Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro

565 570 575

Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser

580 585 590

Asn Gln Val Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val Pro

595 600 605

Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser

610 615 620

Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala

625 630 635 640

Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly

645 650 655

Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser His Gln Gln Ala Gln

660 665 670

Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala

675 680 685

Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn

690 695 700

Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys

705 710 715 720

Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys

725 730 735

Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg

740 745 750

Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val

755 760 765

Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe

770 775 780

Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser

785 790 795 800

Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala

805 810 815

Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala

820 825 830

Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu

835 840 845

Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu

850 855 860

Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala

865 870 875 880

Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile

885 890 895

Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn

900 905 910

Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr

915 920 925

Ala Ser Ala Leu Gly Lys Leu Gln Asn Val Val Asn Gln Asn Ala Gln

930 935 940

Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile

945 950 955 960

Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu Ala

965 970 975

Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln

980 985 990

Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser

995 1000 1005

Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln

1010 1015 1020

Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser

1025 1030 1035

Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr

1040 1045 1050

Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile

1055 1060 1065

Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val

1070 1075 1080

Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu

1085 1090 1095

Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys

1100 1105 1110

Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu

1115 1120 1125

Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe

1130 1135 1140

Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly

1145 1150 1155

Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu

1160 1165 1170

Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln

1175 1180 1185

Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile

1190 1195 1200

Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr

1205 1210 1215

Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly

1220 1225 1230

Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser

1235 1240 1245

Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255 1260

<210> 168

<211> 1261

<212> PRT

<213> 人工序列

<220>

<223> 重组SARS-CoV-2 S多肽

<400> 168

Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr

1 5 10 15

Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser

20 25 30

Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn

35 40 45

Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys

50 55 60

Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala

65 70 75 80

Ser Ile Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr

85 90 95

Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn

100 105 110

Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu

115 120 125

Gly Val Thr Ser Asn His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu

130 135 140

Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser

145 150 155 160

Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn

165 170 175

Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr

180 185 190

Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe

195 200 205

Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr

210 215 220

Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly

225 230 235 240

Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly

245 250 255

Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr

260 265 270

Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys

275 280 285

Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser

290 295 300

Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile

305 310 315 320

Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Lys Phe Ala

325 330 335

Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp

340 345 350

Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr

355 360 365

Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr

370 375 380

Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro

385 390 395 400

Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp

405 410 415

Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys

420 425 430

Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn

435 440 445

Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly

450 455 460

Ser Thr Pro Cys Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu

465 470 475 480

Gln Ser Tyr Gly Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr

485 490 495

Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val

500 505 510

Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn

515 520 525

Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn

530 535 540

Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr

545 550 555 560

Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr

565 570 575

Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr

580 585 590

Ser Asn Gln Val Ala Val Leu Tyr Gln Gly Val Asn Cys Thr Glu Val

595 600 605

Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr

610 615 620

Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly

625 630 635 640

Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala

645 650 655

Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser His Gln Gln Ala

660 665 670

Gln Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly

675 680 685

Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr

690 695 700

Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr

705 710 715 720

Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu

725 730 735

Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn

740 745 750

Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu

755 760 765

Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp

770 775 780

Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro

785 790 795 800

Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu

805 810 815

Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile

820 825 830

Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val

835 840 845

Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala

850 855 860

Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala

865 870 875 880

Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly

885 890 895

Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala

900 905 910

Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser

915 920 925

Thr Ala Ser Ala Leu Gly Lys Leu Gln Asn Val Val Asn Gln Asn Ala

930 935 940

Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala

945 950 955 960

Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Pro Pro Glu

965 970 975

Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu

980 985 990

Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala

995 1000 1005

Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly

1010 1015 1020

Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met

1025 1030 1035

Ser Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val

1040 1045 1050

Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala

1055 1060 1065

Ile Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe

1070 1075 1080

Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr

1085 1090 1095

Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn

1100 1105 1110

Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro

1115 1120 1125

Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr

1130 1135 1140

Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser

1145 1150 1155

Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg

1160 1165 1170

Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu

1175 1180 1185

Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr

1190 1195 1200

Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val

1205 1210 1215

Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys

1220 1225 1230

Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp

1235 1240 1245

Ser Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr

1250 1255 1260

Claims

1.一种冠状病毒(CoV)刺突(S)糖蛋白，所述冠状病毒刺突糖蛋白包含：

其中所述NTD任选地包含选自以下的一个或多个修饰：

(b)在氨基酸132之后1、2、3或4个氨基酸的插入；以及

(c)选自氨基酸5、6、7、13、51、53、56、57、62、63、67、82、125、129、131、132、133、139、143、144、145、177、200、201、202、209、229、233、240、245及其组合的一个或多个氨基酸的突变；

(ii)S2亚基，其中氨基酸973和974是脯氨酸，

其中所述S2亚基任选地包含选自以下的一个或多个修饰：

(b)选自688、703、846、875、937、969、1014、1058、1105和1163及其组合的一个或多个氨基酸的突变；以及

(c)从TMCT中的一个或多个氨基酸的缺失；

其中所述CoV S糖蛋白的氨基酸关于具有SEQ ID NO:2的序列的多肽进行编号。

2.如权利要求1所述的冠状病毒S糖蛋白，所述冠状病毒S糖蛋白包含氨基酸676-685的缺失。

3.如权利要求1所述的冠状病毒S糖蛋白，所述冠状病毒S糖蛋白包含氨基酸702-711的缺失。

4.如权利要求1所述的冠状病毒S糖蛋白，所述冠状病毒S糖蛋白包含氨基酸806-815的缺失。

5.如权利要求1所述的冠状病毒S糖蛋白，所述冠状病毒S糖蛋白包含氨基酸775-793的缺失。

6.如权利要求1所述的冠状病毒S糖蛋白，所述冠状病毒S糖蛋白包含所述NTD的氨基酸1-292的缺失。

7.如权利要求1所述的冠状病毒S糖蛋白，所述冠状病毒S糖蛋白包含跨膜和胞质尾(TMCT)的氨基酸1201-1260的缺失。

8.如权利要求1所述的冠状病毒S糖蛋白，所述冠状病毒S糖蛋白包含选自SEQ ID NO:85-89、105、106和112-115、164-168的氨基酸序列，或由所述氨基酸序列组成。

9.如权利要求1至8中任一项所述的冠状病毒S糖蛋白，所述冠状病毒S糖蛋白包含信号肽，任选地其中所述信号肽包含SEQ ID NO:5或SEQ ID NO:117的氨基酸序列。

10.如权利要求1至9中任一项所述的冠状病毒S糖蛋白，所述冠状病毒S糖蛋白包含C末端融合蛋白。

11.如权利要求10所述的冠状病毒S糖蛋白，其中所述C末端融合蛋白是六组氨酸标签。

12.如权利要求10所述的冠状病毒S糖蛋白，其中所述C末端融合蛋白是折叠子。

13.如权利要求12所述的冠状病毒S糖蛋白，其中所述折叠子具有对应于SEQ ID NO:68的氨基酸序列。

14.如权利要求1-13中任一项所述的冠状病毒S糖蛋白，其中ΔHcal是野生型CoV S糖蛋白(SEQ ID NO:2)的ΔHcal的至少2倍。

15.如权利要求1-14中任一项所述的冠状病毒S糖蛋白，其中所述S2亚基、所述NTD、所述RBD和所述SD1/2各自与具有SEQ ID NO:2的氨基酸序列的CoV S糖蛋白的相应亚基或结构域95％相同。

16.如权利要求1-14中任一项所述的冠状病毒S糖蛋白，其中所述S2亚基、所述NTD、所述RBD和所述SD1/2各自与具有SEQ ID NO:2的氨基酸序列的CoV S糖蛋白的相应亚基或结构域97％相同。

17.如权利要求1-14中任一项所述的冠状病毒S糖蛋白，其中所述S2亚基、所述NTD、所述RBD和所述SD1/2各自与具有SEQ ID NO:2的氨基酸序列的CoV S糖蛋白的相应亚基或结构域99％相同。

18.如权利要求1-14中任一项所述的冠状病毒S糖蛋白，其中所述S2亚基、所述NTD、所述RBD和所述SD1/2各自与具有SEQ ID NO:2的氨基酸序列的CoV S糖蛋白的相应亚基或结构域99.5％相同。

19.一种分离的核酸，所述分离的核酸编码如权利要求1-18中任一项所述的S糖蛋白。

20.一种载体，所述载体包含如权利要求19所述的核酸。

21.一种纳米颗粒，所述纳米颗粒包含如权利要求1-18中任一项所述的冠状病毒S糖蛋白。

22.如权利要求21所述的纳米颗粒，其中所述纳米颗粒的Zavg直径在约20nm与约35nm之间。

23.如权利要求21所述的纳米颗粒，其中所述纳米颗粒的多分散性指数为约0.2至约0.45。

24.一种细胞，所述细胞表达如权利要求1-18中任一项所述的冠状病毒S糖蛋白。

25.一种疫苗组合物，所述疫苗组合物包含如权利要求1-18中任一项所述的冠状病毒S糖蛋白和医药上可接受的缓冲液。

26.如权利要求25所述的疫苗组合物，所述疫苗组合物包含佐剂。

27.如权利要求26所述的疫苗组合物，其中所述佐剂包含至少两种iscom颗粒，其中：

第一iscom颗粒包含皂树(Quillaja Saponaria Molina)的级分A，而不包含皂树的级分C；并且

第二iscom颗粒包含皂树的级分C，而不包含皂树的级分A。

28.如权利要求27所述的疫苗组合物，其中皂树的级分A和皂树的级分C分别占在所述佐剂中皂树的级分A和皂树的级分C的重量之和的按重量计约85％和按重量计约15％。

29.如权利要求27所述的疫苗组合物，其中皂树的级分A和皂树的级分C分别占在所述佐剂中皂树的级分A和皂树的级分C的重量之和的按重量计约92％和按重量计约8％。

30.如权利要求26所述的疫苗组合物，其中将所述佐剂以约50μg的剂量施用。

31.一种在受试者中刺激针对SARS-CoV-2的免疫反应的方法，所述方法包括施用如权利要求25-30中任一项所述的疫苗组合物。

32.如权利要求31所述的方法，其中向所述受试者在第0天施用第一个剂量，并且在第21天施用加强剂量。

33.如权利要求31所述的方法，其中向所述受试者施用约3μg至约25μg的冠状病毒S糖蛋白。

34.如权利要求31所述的方法，其中向所述受试者施用约5μg的冠状病毒S糖蛋白。

35.如权利要求31-34中任一项所述的方法，其中将所述疫苗组合物肌内施用。

36.如权利要求31、33、34和35中任一项所述的方法，其中施用单剂量的所述疫苗组合物。

37.如权利要求31-35中任一项所述的方法，其中施用多剂量的所述疫苗组合物。

38.如权利要求31-37中任一项所述的方法，其中将所述疫苗组合物与流感糖蛋白共同施用。

39.一种免疫原性组合物，所述免疫原性组合物包含：

(i)含有如权利要求1-18中任一项所述的冠状病毒S(CoV S)糖蛋白和非离子型洗涤剂核心的纳米颗粒；

(ii)医药上可接受的缓冲液；以及

(iii)皂苷佐剂。

40.如权利要求39所述的免疫原性组合物，所述免疫原性组合物包含约3μg至约25μg的CoV S糖蛋白。

41.如权利要求40所述的免疫原性组合物，所述免疫原性组合物包含约5μg的CoV S糖蛋白。

42.如权利要求39所述的免疫原性组合物，其中所述皂苷佐剂包含至少两种iscom颗粒，其中：

第一iscom颗粒包含皂树的级分A，而不包含皂树的级分C；并且

第二iscom颗粒包含皂树的级分C，而不包含皂树的级分A。

43.如权利要求42所述的免疫原性组合物，其中分别地，皂树的级分A占在所述佐剂中皂树的级分A和皂树的级分C的重量之和的按重量计50％-96％，并且皂树的级分C占剩余部分。

44.如权利要求42所述的免疫原性组合物，其中皂树的级分A和皂树的级分C分别占在所述佐剂中皂树的级分A和皂树的级分C的重量之和的按重量计约85％和按重量计约15％。

45.如权利要求39所述的免疫原性组合物，所述免疫原性组合物包含约50μg的皂苷佐剂。

46.如权利要求39所述的免疫原性组合物，其中所述非离子型洗涤剂选自聚山梨醇酯20(PS20)、聚山梨醇酯40(PS40)、聚山梨醇酯60(PS60)、聚山梨醇酯65(PS65)和聚山梨醇酯80(PS80)。

47.一种在受试者中刺激针对SARS-CoV-2或异源SARS-CoV-2毒株的免疫反应的方法，所述方法包括施用如权利要求39-46中任一项所述的免疫原性组合物。

48.如权利要求47所述的方法，其中施用约3μg至约25μg的CoV S糖蛋白。

49.如权利要求47或48所述的方法，其中施用约5μg的CoV S糖蛋白。

50.如权利要求47-49中任一项所述的方法，其中所述皂苷佐剂包含至少两种iscom颗粒，其中：

第一iscom颗粒包含皂树的级分A，而不包含皂树的级分C；并且

第二iscom颗粒包含皂树的级分C，而不包含皂树的级分A。

51.如权利要求50所述的方法，其中分别地，皂树的级分A占在所述佐剂中皂树的级分A和皂树的级分C的重量之和的按重量计50％-96％，并且皂树的级分C占剩余部分。

52.如权利要求50或权利要求51所述的方法，其中皂树的级分A和皂树的级分C分别占在所述佐剂中皂树的级分A和皂树的级分C的重量之和的按重量计约85％和按重量计约15％。

53.如权利要求47-52中任一项所述的方法，所述方法包括施用约50μg的所述皂苷佐剂。

54.如权利要求47-53中任一项所述的方法，其中所述非离子型洗涤剂选自聚山梨醇酯20(PS20)、聚山梨醇酯40(PS40)、聚山梨醇酯60(PS60)、聚山梨醇酯65(PS65)和聚山梨醇酯80(PS80)。

55.如权利要求47-54中任一项所述的方法，其中向所述受试者在第0天施用第一个剂量，并且在第21天施用加强剂量。

56.如权利要求47-55中任一项所述的方法，其中施用单剂量的所述免疫原性组合物。

57.如权利要求47-56中任一项所述的方法，所述方法进一步包括施用与如权利要求47所述的免疫原性组合物不同的第二免疫原性组合物。

58.如权利要求57所述的方法，其中所述第二免疫原性组合物包含编码SARS-CoV-2刺突糖蛋白的mRNA、编码SARS-CoV-2刺突糖蛋白的质粒DNA、编码SARS-CoV-2刺突糖蛋白的病毒载体或灭活的SARS-CoV-2病毒。

59.如权利要求47-58中任一项所述的方法，其中所述异源SARS-CoV-2毒株选自B.1.1.7SARS-CoV-2毒株、B.1.351SARS-CoV-2毒株、P.1SARS-CoV-2毒株、B.1.617.2SARS-CoV-2毒株、B.1.525SARS-CoV-2毒株、B.1.526SARS-CoV-2毒株、B.1.617.1SARS-CoV-2毒株、C.37SARS-CoV-2毒株、B.1.621SARS-CoV-2毒株和Cal.20CSARS-CoV-2毒株。

60.如权利要求47-59中任一项所述的方法，其中所述免疫原性组合物用于预防冠状病毒病-19(COVID-19)的功效为至少约50％、至少约55％、至少约60％、至少约65％、至少约70％、至少约75％、至少约80％、至少约85％、至少约90％、至少约91％、至少约92％、至少约93％、至少约94％、至少约95％、至少约96％、至少约97％、至少约98％、至少约99％或约100％，所述功效在施用所述免疫原性组合物后持续长达约2个月、长达约2.5个月、长达约3个月、长达约3.5个月、长达约4个月、长达约4.5个月、长达约5个月、长达约5.5个月、长达约6个月、长达约6.5个月、长达约7个月、长达约7.5个月、长达约8个月、长达约8.5个月、长达约9个月、长达约9.5个月、长达约10个月、长达约10.5个月、长达约11个月、长达约11.5个月或长达约12个月。

61.如权利要求47-60中任一项所述的方法，其中所述免疫原性组合物用于预防冠状病毒病-19(COVID-19)的功效为从约50％至约99％、从约50％至约95％、从约50％至约90％、从约50％至约85％、从约50％至约80％、从约60％至约99％、从约60％至约95％、从约60％至约90％、从约60％至约85％、从约60％至约80％、从约40％至约99％、从约40％至约95％、从约40％至约90％、从约40％至约85％、从约40％至约80％、从约40％至约75％、从约40％至约70％、从约40％至约65％、从约40％至约55％或从约40％至约50％，所述功效在施用所述免疫原性组合物后持续长达约2个月、长达约2.5个月、长达约3个月、长达约3.5个月、长达约4个月、长达约4.5个月、长达约5个月、长达约5.5个月、长达约6个月、长达约6.5个月、长达约7个月、长达约7.5个月、长达约8个月、长达约8.5个月、长达约9个月、长达约9.5个月、长达约10个月、长达约10.5个月、长达约11个月、长达约11.5个月或长达约12个月。

62.如权利要求47-61中任一项所述的方法，其中所述COVID-19是轻度COVID-19。

63.如权利要求47-61中任一项所述的方法，其中所述COVID-19是中度COVID-19。

64.如权利要求47-61中任一项所述的方法，其中所述COVID-19是重度COVID-19。

65.一种诱导针对异源SARS-CoV-2毒株的保护性免疫反应的方法，所述方法包括向受试者施用包含具有SEQ ID NO:87的氨基酸序列的冠状病毒S(CoV S)糖蛋白和非离子型洗涤剂核心的纳米颗粒、医药上可接受的缓冲液以及(iii)皂苷佐剂，其中所述异源SARS-CoV-2毒株具有与SEQ ID NO:1的SARS-CoV-2糖蛋白相比含有约1至约60个修饰的SARS-CoV-2S糖蛋白。

66.如权利要求65所述的方法，其中所述异源SARS-CoV-2毒株具有与SEQ ID NO:1的SARS-CoV-2糖蛋白相比含有约1至约20个修饰、约1至约10个修饰、约10至约20个修饰、10至约30个修饰、约10至约40个修饰、10至约50个修饰、10至约60个修饰、20至约60个修饰、20至约50个修饰、约20至约40个修饰、约5至约15个修饰或约5至约10个修饰的SARS-CoV-2S糖蛋白。