CN106687590A

CN106687590A - 黄热病毒病毒样颗粒

Info

Publication number: CN106687590A
Application number: CN201580048888.9A
Authority: CN
Inventors: 赤畑涉; 上野隆司
Original assignee: VLP Therapeutics Inc
Current assignee: VLP Therapeutics Inc
Priority date: 2014-09-11
Filing date: 2015-09-10
Publication date: 2017-05-17
Anticipated expiration: 2035-09-10
Also published as: SG11201701669PA; AR101814A1; CN106687590B; BR112017004770A8; JP2017528139A; KR20170055513A; EP3191589A4; TWI695842B; PH12017500450A1; BR112017004770A2; CA2960102A1; KR102532832B1; AU2015313650B2; TW201612190A; WO2016038895A1; US10098943B2; AU2015313650A1; CA2960102C; JP6942309B2; MX2017003117A

Abstract

本申请提供了包含一种或多种黄热病毒结构蛋白的病毒样颗粒，以及包含所述病毒样颗粒的组合物或疫苗，其在预防或治疗黄热病毒感染中的用途。所述黄热病毒结构蛋白含有包膜区域中的至少一个氨基酸变化。黄热病毒的实例含有登革病毒。

Description

黄热病毒病毒样颗粒

技术领域

本申请涉及包含一种或更多种黄热病毒结构蛋白的病毒样颗粒，以及包含其的组合物或疫苗，其在医药中、尤其是黄热病毒感染的预防或治疗中的用途。

背景技术

黄热病毒包含超过70种不同的病毒，其中许多为节肢动物媒介且由蚊子或蜱传播。

黄热病毒是黄热病毒科中的病毒的一个属。该属包括西尼罗河病毒(WNV)、登革病毒(DENV)、蜱媒脑炎病毒(TBEV)、黄热病病毒(YFV)、日本脑炎(JEV)、丙型肝炎病毒(HCV)以及几种其他可能引起脑炎或出血性疾病的病毒。

登革热是一种由该黄热病毒引起的蚊媒疾病且已传播至大多数热带和许多亚热带地区。该疾病由四种密切相关的病毒(包括亚型I至IV的登革病毒1，包括亚型亚洲I、亚洲II、全球、美洲和美洲/亚洲的登革病毒2，包括亚型I至IV的登革病毒3，以及包括亚型I至III的登革病毒4)引起。尽管登革热就全球疾病发病率而言是最重要的黄热病毒，但针对该病毒的疫苗的开发和使用迄今已经受到疫苗相关不良事件(诸如感染的免疫增强和对于诱导长效保护免疫应答以同时对抗全部四种登革血清型的需求)的理论风险的阻碍。

没有有效的登革热治疗且针对登革热的预防目前仅限于媒介控制措施。因此，登革热疫苗将代表疾病控制中的重大进步。

尽管没有许可的登革热疫苗可用，但目前在临床研究中评估了几种疫苗候选物。WHO指明，全球越来越多的登革热流行病令人担忧，且迫切需要一种安全且有效的疫苗。

(http://www.who.int/immunization/research/development/dengue_vaccines/en/) and Vaccine 30 (2012) 4301-4306)。

病毒样颗粒(VLPs)是多蛋白结构，其模拟真实的天然病毒的组织和构象，但是非感染性的，因为它们不含任何病毒基因组，潜在地产生更安全的疫苗候选物。目前在世界范围内销售了几种预防性的基于VLP的疫苗：GlaxoSmithKline's Engerix^R(乙型肝炎病毒)和Cervarix^R(人乳头瘤病毒)，以及Merck and Co., Inc.的Recombivax HB^R(乙型肝炎病毒)和Gardasil^R(人乳头瘤病毒)是一些实例。其他基于VLP的疫苗候选物在临床试验中或正在经历临床前评估，诸如流感病毒、细小病毒、诺瓦克病毒和各种嵌合VLP。许多其他仍然局限于小规模基础研究，尽管它们在临床前测试中取得了成功。在过程控制、监测和优化的背景下讨论大规模VLP生产的影响。从而鉴定并讨论了主要的上下游技术挑战。针对治疗或预防性接种疫苗，伴随着来自临床试验的最新结果和基于嵌合VLP的技术的近来发展，成功的基于VLP的疫苗畅销品(blockbusters)短暂地出现。

迄今为止，已经为感染人类和其他动物的超过30种不同病毒生产了基于VLP的疫苗。实例包括AAV(腺相关病毒)、H5N3(禽流感)、BFDV(虎皮鹦鹉掉羽症病毒)、BTV(蓝舌病毒)、埃博拉病毒、肠道病毒71、GHPV(鹅出血性多瘤病毒)、HBV(乙型肝炎病毒)、HCV(丙型肝炎病毒)、HDV(丁型肝炎病毒)、HEV(戊型肝炎病毒)、HIV、HPV(人乳头瘤病毒)、IBDV(传染性囊病病毒)、甲型流感、甲型流感H1N1、甲型流感H3N2、JC多瘤病毒、马堡病毒(Margurg)、MS2、IPCV(印度花生丛病毒)、NDV(新城疫病毒)、No(诺如病毒)、Nv(诺瓦克病毒)、PhMV(酸浆花叶病毒(Physalis mottle virus))、多瘤病毒、PPV(猪细小病毒)，RHDV(兔出血疾病病毒)、轮状病毒、SARS、SIV(猿猴免疫缺陷病毒)、SV40(猿猴病毒40)、SVDV(猪水疱病病毒)等等。(Expert Rev. Vaccines 9(10), 1149-1176, 2010)。

[引文列表]

[非专利文献]

[NPTL1]Vaccine 30 (2012) 4301-4306

[NPTL2] Expert Rev. Vaccines 9(10), 1149-1176, 2010。

[发明概述]

在第一个方面，本申请提供包含一种或更多种黄热病毒结构蛋白的病毒样颗粒，其中所述黄热病毒结构蛋白的包膜区域含有氨基酸序列中的至少一个变化。

在第二个方面，本申请提供核酸分子，其包含编码本申请的第一个方面中提供的病毒样颗粒中含有的黄热病毒结构蛋白的核苷酸序列或由其组成。

在第三个方面，本申请提供组合物或疫苗，其包含第一个方面中提供的病毒样颗粒和/或第二个方面中提供的核酸分子。

在第四个方面，本申请提供产生抗体或包含中和抗体的抗血清的方法，其包括将本申请的第一个方面中提供的病毒样颗粒和/或本申请的第二个方面中提供的核酸分子与哺乳动物接触。

在第五个方面，本申请提供治疗或预防黄热病毒感染的方法或在哺乳动物中诱导和/或增强针对黄热病毒的免疫应答的方法，其包括将本申请的第三个方面中提供的组合物施用于哺乳动物。

在第六个方面，本申请提供产生本申请的第一个方面中提供的病毒样颗粒的方法，其包括：培养用编码病毒样颗粒中含有的至少一种黄热病毒结构蛋白的基因转染的细胞；以及从细胞培养物回收病毒样颗粒。该方面可进一步包括制备包含编码病毒样颗粒中包含的至少一种黄热病毒结构蛋白的核苷酸序列的基因的步骤。

在第七个方面，本申请提供用于诊断哺乳动物的黄热病毒感染的方法或试剂盒中的病毒样颗粒。

[附图简述]

[图1]

图1显示登革病毒基因组的构建以及登革VLP构建体的一个实例。

[图2]

图2显示通过Western印迹测量的上清液中的包膜蛋白。将包含登革2型病毒prM和包膜蛋白的病毒结构蛋白的表达载体，或包含登革2型病毒prM和登革2型和登革1型的嵌合体(其中登革病毒2型包膜蛋白的C端区域被置换为对应的登革病毒1型C端区域(SEQ ID NO:8的199aa-676aa))的病毒结构蛋白的表达载体，或含有嵌合包膜区域的病毒结构蛋白(其中引入嵌合包膜区域中一个氨基酸变化(F108K，在SEQ ID NO: 10的289aa))的表达载体以及登革病毒1型衣壳蛋白的表达载体转染至293F细胞。针对培养上清液进行使用针对登革包膜蛋白的抗体的Western印迹。

[图3]

图3显示Western印迹的结果。将包含登革1型病毒prM和具有或不具有变化的包膜(野生型、F108A、K203M、K246M、F108A+K246M或F108+K203N)的病毒结构蛋白的表达载体和登革1型衣壳蛋白的表达载体一起转染至293F细胞。针对培养上清液进行使用针对登革包膜蛋白的抗体的Western印迹。

[图4]

图4显示Western印迹的结果。将包含登革1型病毒prM和具有或不具有变化的包膜(野生型、F108A、K203M或F108+K203N)的病毒结构蛋白的表达载体转染至293F细胞。针对培养上清液进行使用针对登革包膜蛋白的抗体的Western印迹。

[图5]

图5显示负染的VLP(F108+K203N)的电子显微(EM)显影。

[实施方案的描述]

(1)包含一种或更多种黄热病毒结构蛋白的病毒样颗粒

在第一个方面，本申请提供包含一种或更多种黄热病毒结构蛋白的病毒样颗粒，其中包膜区域中的至少一个氨基酸从其天然存在的结构变化。

黄热病毒可以是西尼罗河病毒、登革病毒、蜱媒脑炎病毒、黄热病病毒、日本脑炎和丙型肝炎(HCV)。优选的黄热病毒为登革病毒。登革病毒可以是包括亚型I至IV的登革病毒1，包括亚型亚洲I、亚洲II、全球、美洲和美洲/亚洲的登革病毒2，包括亚型I至IV的登革病毒3，以及包括亚型I至III的登革病毒4。在说明书和权利要求中，术语“黄热病毒结构蛋白或其片段”是指衍生自黄热病毒的任何基于肽的序列，其可被对象的免疫系统识别，和/或刺激对象中的细胞介导的免疫应答和/或刺激对象中抗体的生成。

如图1中所显示，登革病毒结构蛋白由衣壳蛋白、前体膜蛋白和包膜蛋白组成。在该具体实施方案中，VLP包含那些结构蛋白中的至少一种，以及优选前体膜蛋白(prM)和包膜蛋白。登革病毒结构蛋白可进一步包含对应于prM序列的起始密码子和信号序列至氨基末端的氨基酸。

包含至少一个氨基酸变化的区域可优选地在SEQ ID NO: 21(其包含起始密码子M和SEQ ID NO: 17的信号肽，随后为prM和登革病毒1型的包膜区域(WestPac毒株，GenBank登录号：U88535))的蛋白的氨基酸位置182和氨基酸位置676之间，更优选在氨基酸位置271和氨基酸位置302之间，尤其是在氨基酸位置280和氨基酸位置291之间，特别是位置289，或在当包含起始密码子M、信号肽、随后的prM和黄热病毒的包膜区域的蛋白或其片段的氨基酸序列与由SEQ ID NO: 21表示的氨基酸序列比对时，确定为上述鉴定的位置的位置之间。

登革病毒1型的包膜区域的氨基酸序列由SEQ ID NO: 20表示。因此，包含至少一个氨基酸变化的区域可在该序列中，优选地在SEQ ID NO: 20的蛋白的氨基酸位置90和氨基酸位置121之间，尤其是在氨基酸位置99和氨基酸位置110之间，特别是位置108，或在当黄热病毒的包膜或其片段的氨基酸序列与SEQ ID NO: 20比对时，确定为上述鉴定的位置的位置之间。

如本文所用，“对应于…的位置”或记载核苷酸或氨基酸位置“对应于”公开序列(诸如序列表中所记载)中的核苷酸或氨基酸位置，是指在使用标准比对算法与公开的序列比对以最大化同一性被鉴定的核苷酸或氨基酸位置。通过比对所述序列，本领域技术人员可鉴定对应的残基。

各种黄热病毒诸如登革病毒1-4型的病毒结构蛋白已经被鉴定，且可在各种公开数据库诸如GenBank数据库获得。例如，登革病毒1型(WestPac毒株)：登录号U88535、登革病毒2型(S1疫苗毒株)：登录号M19197、登革病毒3型(毒株Singapore 8120/95)：登录号AY766104以及登革病毒4型(毒株ThD4_0476_97)：登录号Y618988。根据本申请，黄热病毒诸如登革病毒的包膜蛋白可自数据库获得，且与SEQ ID NO: 20比对以鉴定对应于SEQ IDNO: 20的氨基酸位置90和氨基酸位置121之间位置的位置，诸如对应于SEQ ID NO: 20的氨基酸位置108的位置。

除了在包膜区域中包含至少一个氨基酸变化之外，病毒样颗粒中含有的黄热病毒结构蛋白可以是天然存在的病毒结构蛋白或其修饰的蛋白。在一个实施方案中，修饰的蛋白与包括prM和包膜蛋白的天然存在的病毒结构蛋白具有至少70%、75%、80%、85%、90%、95%或98%氨基酸序列同一性。在一个实施方案中，修饰的蛋白为突变体，其中向包括prM和包膜区域的天然存在的病毒结构蛋白中缺失、取代和/或添加至多10%的氨基酸。序列同一性可通过常规方法确定。

根据本申请，可使用上面讨论的一种或更多种黄热病毒结构蛋白或其片段，只要它们自动组装为颗粒结构。例如，当培养表达编码登革病毒的prM和包膜蛋白的基因的真核细胞时，所述蛋白由细胞生成并组装以提供VLP，且VLP可以从细胞培养上清液收集。

本申请通过提供VLP、编码VLP的载体以及特异性结合至抗原的抗体(和抗体样分子，包括适体和肽)连同其(单独或组合)在黄热病毒感染的预防或治疗中的用途来解决了一种或更多种上述需求。

如本说明书和权利要求所用，术语“抗体”是指能够结合至表位或抗原决定簇的分子。该术语涵盖全部抗体和其抗原结合片段，包括单链抗体。此类抗体可包括人类抗原结合抗体片段且包括，但不限于，Fab、Fab'和F(ab')2、Fd、单链Fvs(scFv)、单链抗体、二硫键连接的Fvs(sdFv)以及包含VL或VH区域的片段。所述抗体可来自任何动物来源，包括鸟类和哺乳动物。优选地，所述抗体为哺乳动物的，例如人类、鼠、兔、山羊、豚鼠、骆驼、马等，或其他合适的动物，例如鸡。如本文所用，“人类”抗体包括具有人类免疫球蛋白的氨基酸序列的抗体，且包括分离自人类免疫球蛋白文库或分离自转基因有一种或更多种人类免疫球蛋白且不表达内源性免疫球蛋白的动物的抗体，如描述于例如美国专利号5,939,598，其公开内容以其整体并入本文。

黄热病毒结构蛋白可以是天然存在的蛋白或天然存在的蛋白的修饰的蛋白或天然存在的蛋白的片段或修饰的肽。修饰的蛋白可以是天然存在的病毒结构蛋白的片段。

在一个实施方案中，衍生自黄热病毒结构蛋白的修饰的蛋白与天然存在的蛋白具有至少70%、75%、80%、85%、90%、95%或98%氨基酸序列同一性。在一个实施方案中，衍生自黄热病毒的修饰的蛋白为突变体，其中基于衍生自黄热病毒的天然存在的病毒结构蛋白，缺失、取代和/或添加至多10%的氨基酸。

在一个实施方案中，本申请提供病毒样颗粒，其包含具有由SEQ ID No.2、4、8、10、14和16中任一者表示的氨基酸序列的黄热病毒结构蛋白。

那些序列各自表示含有以下区域的蛋白：起始密码子：M(1aa)、信号序列(2-15aa)、pr序列(16-106aa)、M序列(107-181aa)和包膜区域(182-676aa)。在一个实施方案中，可提供病毒样颗粒，其包含具有prM的区域和SEQ ID No.2、4、8、10、14和16的包膜区域的病毒结构蛋白，即包含那些蛋白的位置16至位置676的氨基酸序列。

修饰的黄热病毒结构蛋白与由SEQ ID No.2、4、8、10、14和16中任一者表示的氨基酸序列，或那些序列之一的位置16至位置676的氨基酸序列具有至少70%、75%、80%、85%、90%、95%或98%氨基酸序列同一性。此外，修饰的黄热病毒结构蛋白可以是突变体，其中基于具有由SEQ ID No.2、4、8、10、14和16中任一者表示的氨基酸序列或那些序列任一的位置16至位置676的氨基酸序列的黄热病毒病毒结构蛋白，缺失、取代和/或添加至多10%的氨基酸。

(2)核苷酸和载体

在第二个方面，本申请提供核酸分子，包含编码如本申请的第一个方面中提供的病毒样颗粒的核苷酸序列或由其组成。

在一个实施方案中，本申请提供核酸分子，其包含核苷酸序列，其编码提供如上所述的病毒样颗粒的黄热病毒结构蛋白。

在一个实施方案中，本申请提供包含如上所述的核酸分子的表达载体，其中所述载体任选地包含可操作地连接至该核酸分子的表达控制序列。

表达控制序列的实例包括，但不限于，启动子诸如CMV启动子、噬菌体lambda PL启动子、大肠杆菌lac、phoA和tac启动子、SV40早期和晚期启动子和逆转录病毒LTR的启动子。

在一个实施方案中，本申请提供用于黄热病毒结构蛋白的表达载体，其由SEQ IDNo.1、3、7、9、13和15中的任一者表示的核苷酸序列组成。

在一个实施方案中，本申请提供核酸分子，其从具有由SEQ ID No.1、3、7、9、13和15中的任一者表示的核苷酸序列的核酸分子修饰。修饰的核酸序列与具有由SEQ ID No.1、3、7、9、13和15中的任一者表示的核苷酸序列的核酸分子可具有至少70%、75%、80%、85%、90%、95%或98%核苷酸序列同一性。此外，修饰的核酸分子可以是突变体，其中基于具有由SEQ ID No.1、3、7、9、13和15中的任一者表示的核苷酸序列的核酸分子，缺失、取代和/或添加至多10%的氨基酸。

(3)组合物或疫苗

在第三个方面，本申请提供组合物或疫苗，其包含本申请的第一个方面中提供的病毒样颗粒和/或本发明第二个方面中提供的核酸分子。

在一个实施方案中，本申请提供包含如上所述的黄热病毒病毒样颗粒或如上所述的核酸分子的组合物。

所述组合物可进一步包含药学上可接受的载体和/或佐剂。佐剂的实例包括，但不限于Ribi溶液(Sigma佐剂系统，Sigma-Aldrich)。

本申请的药物组合物可含有单一活性成分或两种或更多种活性成分的组合，只要它们不与本申请的目标相反。例如，细胞因子包括趋化因子、针对细胞因子的抗体，诸如抗-TNF抗体(例如，英利昔单抗(infliximab)、阿达木单抗(adalimumab))、抗-VEGF抗体(例如，贝伐珠单抗(bevacizumab)和兰尼单抗(ranibizumab))、细胞因子受体拮抗剂，诸如抗HER2抗体(例如，曲妥珠单抗(Trastuzumab))、抗EGF受体抗体(例如，西妥昔单抗(Cetuximab))、抗VEGF适体(例如，哌加他尼(Pegaptanib))和免疫调节剂诸如环孢霉素、他克莫司(tacrolimus)和乌苯美司(ubenimex)可用于组合疗法。

在多种活性成分的组合中，考虑到其疗效和安全性，各成分的含量可适当地增加或减少。

如本文所用的术语“组合”意指两种或更多种活性成分以单一实体或剂量的形式同时施用于患者，或者那些活性成分作为分开的实体同时或没有具体时间限制地依次施用于患者，其中此施用优选地在相同时间在体内提供两种成分的治疗有效水平。

在一个实施方案中，所述组合物是疫苗组合物，包括DNA疫苗。在一个实施方案中，本申请提供的DNA疫苗包含含有寡核苷酸的CpG。

(4)产生抗体的方法

在第四个方面，本申请提供产生针对黄热病毒的抗体或含有针对黄热病毒的中和抗体的抗血清的方法，其包括将本申请的第一个方面中提供的病毒样颗粒和/或本申请的第二个方面中提供的核酸分子与哺乳动物接触。

在该方面中产生的抗体可通过向哺乳动物施用该抗体而用于在哺乳动物中针对黄热病毒导致的病原体的被动免疫，并且因此预防哺乳动物免于黄热病毒感染或治疗哺乳动物的由黄热病毒感染导致的疾病或病况。

本申请的第四个方面中产生的抗体可使用常规技术人源化。因此，在一个实施方案中，本申请的第四个方面中提供的方法进一步包括将非人类哺乳动物产生的抗体人源化的步骤。该方面提供的抗体或人源化抗体可用于预防人类对象免于黄热病毒感染或治疗对象中由黄热病毒感染导致的疾病或病况。

根据该方面产生的抗体可体外用于从免疫细胞诸如衍生自患者的B细胞和T细胞选择亚群，其然后再施用于患者。

抗血清可通过常规方式取得。血液样品取自免疫的非人类动物，且将血液处理以便获得抗血清，即含有抗体的血液的液体组分。非人类哺乳动物优选选自大鼠、小鼠、仓鼠、猪、兔、马、驴、山羊、绵羊、豚鼠、羊驼和非人类灵长类动物(例如，黑猩猩)。

(5)治疗对象中由黄热病毒感染导致疾病或预防对象免于黄热病毒感染的方法

在第五个方面，治疗对象中由黄热病毒感染诸如登革热导致的疾病或病况的方法，其中将第一个方面中提供的病毒样颗粒、第二个方面中提供的核酸分子或第三个方面中提供的组合物施用于对象。通过将上面列举的VLP、核酸分子或组合物施用于对象，针对黄热病毒的免疫应答可被增强，因此，可有效地治疗由黄热病毒感染导致的疾病或病况。在该方面，可将VLP、核酸分子或组合物局部地施用于患者的受影响的器官或全身性地施用于患者。

预防对象免于黄热病毒感染或免于由黄热病毒感染导致的疾病的方法，其包括将第一个方面的病毒样颗粒、第二个方面中提供的核酸分子或第三个方面的组合物施用于对象。由黄热病毒导致的疾病可以是登革热。

根据本申请，病毒样颗粒也可应用于免疫疗法。VLP可离体应用至衍生自患者的细胞或人类细胞系，其随后施用于患者。

(6)产生病毒样颗粒的方法

在第六个方面，本申请提供产生本申请的第一个方面中提供的病毒样颗粒的方法，其包括：培养细胞，所述细胞表达编码病毒结构蛋白的基因；以及从细胞培养物回收病毒样颗粒。

各种宿主-载体系统可用于病毒样颗粒的表达。真核细胞可用于本申请的第四个方面提供的方法。真核细胞的实例包括，但不限于，昆虫细胞(例如，sf9细胞、H5细胞)、酵母细胞(例如，酿酒酵母(S. cerevisiae))和哺乳动物细胞(例如，CHO细胞、人类胚胎肾脏(HEK)293F细胞)。用于本申请的第二个方面提供的方法的载体包含编码待表达的病毒样颗粒的核酸分子。可使用常规方法(例如，脂质转染、电穿孔)用载体转染细胞。技术人员可根据采用的细胞选择培养基或添加DNA甲基转移酶抑制剂和组蛋白去乙酰酶抑制剂诸如丁酸钠。转染后，病毒样颗粒可产生于细胞和/或培养上清液中。病毒样颗粒可回收自培养上清液并使用超速离心纯化。

本申请的病毒样颗粒不复制，因此具有高安全性。

(7)诊断黄热病毒感染的试剂盒或方法

在第七个方面，本申请提供用于诊断哺乳动物的黄热病毒感染的方法或试剂盒中的病毒样颗粒。通过使用病毒样颗粒，可以产生能够检测对于黄热病毒特异性的抗体的酶联免疫吸附测定(ELISA)诊断试剂盒。

本申请将参考以下实施例进行详细描述，然而该实施例并不意图限制本申请的范围。

[实施例1]

包含登革病毒1-2型的蛋白或片段的病毒样颗粒的制备

登革病毒1和2型病毒结构蛋白用作野生型病毒结构蛋白。在病毒结构蛋白的包膜区域中引入至少一个变化。即，在登革病毒1型包膜蛋白(SEQ ID NO: 20)的位置108的氨基酸Phe变成Ala(F108A)，在登革病毒1型包膜蛋白的位置203的氨基酸Lys变成Asn(K203N)(包膜区域：SEQ ID NO: 4的172-676aa)，在位置246的氨基酸Asn变成Met(K246M)。在本实施例中，包膜区域对应于SEQ ID NO: 2的172-676aa，SEQ ID NO: 20的位置108对应于SEQ IDNO: 2的位置289，SEQ ID NO: 20的位置203对应于SEQ ID NO: 2的位置384，且SEQ ID NO:20的位置246对应于SEQ ID NO: 2的位置427。在本实施例中，该包膜区域中的突变使用起始自包膜区域的数字表述。

为了在哺乳动物细胞中表达病毒结构蛋白，混合DENV2 prM E表达载体和DENV2衣壳蛋白表达载体各20ug并转染至293F细胞。将衣壳蛋白载体进行R85A和K86A突变以表达更好的产量(SEQ ID NO: 11和12)。为了更好地表达DENV2 prM E，C端DENV2 Env区域(199aa-676aa)被置换为对应的DENV1 Env区域以提供嵌合体蛋白(SEQ ID NO.7和8)。为了稳定VLP，Env融合肽区域(F108A)被突变(SEQ ID NO.9和10)。

转染后4天，从转染的细胞培养物收获上清液，并使用特异结合至包膜蛋白的登革病毒(9.F.10)单克隆抗体(sc-70959，Santa Cruz Biotechnology)作为一级抗体以及兔抗小鼠IgG-HRP(sc-358920，Santa Cruz Biotechnology)作为二级抗体，通过Western印迹检查Env表达。结果显示于图2。

如从图2所显示，用DENV2 prM和具有一个变化(F108A)的登革病毒2型和1型包膜蛋白的嵌合体转染的细胞产生较高量的蛋白。上清液中的那些蛋白自动组装以提供颗粒。

为了表达VLP，混合DENV1 prM E表达载体和DENV1衣壳蛋白表达载体各20ug并转染至293F细胞。将衣壳蛋白载体进行K85A和K86A突变以表达更好的产量(SEQ ID NO.5和6)。为了稳定VLP，Env融合肽区域(F108A)(SEQ ID NO.1和2)或碱性氨基酸(K203N)(SEQ IDNO.3和4)或(K246M)被突变。此外，两个氨基酸(F108A+K246M，SEQ ID NO: 13和14)和(F108A+K203N，SEQ ID NO.15和16)也被突变。转染后4天，从转染的细胞培养物收获上清液，并使用登革病毒(9.F.10)的单克隆抗体(sc-70959，Santa Cruz Biotechnology)作为一级抗体以及兔抗小鼠IgG-HRP(sc-358920，Santa Cruz Biotechnology)作为二级抗体，通过Western印迹检查Env表达。结果显示于图3。

如由图3所显示，用DENV prM和具有至少一个变化的包膜蛋白转染的细胞产生较高量的包膜蛋白。上清液中的那些蛋白自动组装以提供颗粒。

[实施例2]

包含登革病毒结构蛋白prM和包膜的病毒样颗粒的制备

使用实施例1中所用的含有prM和修饰的包膜蛋白F108A、K203N或K203N+F108A(分别为SEQ ID NO1、3、15)的登革病毒结构蛋白的表达载体。以与实施例1相同的方式，将20ug的载体转染至293F细胞并培养。在转染后第4天，从转染的细胞培养物收获上清液。使用单克隆抗体(9.F.10,Santa Cruz Biotech)作为一级抗体以及兔抗小鼠IgG-HRP缀合抗体作为二级抗体，进行Western印迹以检测DENV VLP。结果显示于图4。如由图4所显示，用DENV1-prM和具有至少一个变化的包膜蛋白转染的细胞产生较高量的包膜蛋白。

使用0.45μm过滤器过滤上清液以获得病毒样颗粒。病毒样颗粒使用TFF柱浓缩并使用QXL柱(GE Healthcare)纯化以获得纯化的病毒样颗粒。纯化的病毒样颗粒保存于4%甲醛/PBS中。进行负染的病毒样颗粒的电子显微(EM)显影。简言之，将1.0μL的样品置于碳包被的Formvar成膜铜网上并使VLP贴附。然后，将2μL 1% PTA溶液添加至该网，并通过电子显微镜检验该网。结果显示于图5。在上清液中观察到病毒样颗粒。

[实施例3]

针对登革病毒的抗体

实施例2中获得的纯化的VLP (K203N+F108A)被命名为DEN1 VLP并用于本实施例中。将纯化的病毒样颗粒使用离心柱(分子量截止值：100kDa)进一步浓缩以制备用于免疫的病毒样颗粒。然后，通过在第0和4周肌内注射两次而用含铝佐剂(alhydrogel 2%, SergeantAduvants)的PBS中的DEN1 VLP 30μg免疫4只小鼠。

衍生自免疫的小鼠的血清的登革病毒特异性IgG滴度通过ELISA系统测定。测定该血清中针对DENV-1、DENV-2、DENV-3和DENV-4病毒各自的中和抗体。使用以下登革血清型1至4：

DENV-1, Philippine-99St12A毒株

DENV-2, Philippine-00St22A毒株

DENV-3, Philipine-SLMC50毒株

DENV-4, Philippine-SLMC318毒株

JEV- JaOAr S-982毒株。

免疫小鼠血清中抗-DENV中和抗体通过前述灶点减少中和测试(focus reductionneutralization test, FRNT)在Vero细胞上使用1.25 x 10⁸ FFU的Vero-适应的DV 1-4型来检测。终点滴度被计算为测试的最高血清稀释度，其使FFU的数字降低至少50% (FRNT₅₀)。使用在96孔板中100%汇合的Vero细胞。

使用内部抗-登革兔IgG作为一级抗体，且使用抗-兔IgG-HRPO缀合物(102-PD)作为二级抗体。结果概述于下表中。

血清型	D-1	D-2	D-3	D-4	JE
						DENV1 VLP	>10240	1043	604	270	<80
PBS	<80	<80	<80	<80	<80

如表1中所显示，DENV1 VLP显示对登革病毒1、2、3和4型的优异中和效果。考虑到用登革减毒活疫苗获得的中和抗体的FRNT 50值为约1000(Tsuai等人, Journal ofVirology, 2015 89: 7348–7362.)，DENV1 VLP的免疫原性是非常强的并且提供了减毒活疫苗的约10倍的针对登革病毒的中和抗体。

[实施例4]

包含登革病毒样颗粒的药物组合物的制备

根据实施例1制备登革病毒样颗粒。为了制备作为疫苗组合物的药物组合物，将80μg各制备的颗粒与1ml蔗糖磷酸盐溶液(pH 7.2，无内毒素，Teknova，SP缓冲液)混合。

[实施例5]

登革病毒2-4型衍生病毒样颗粒的制备

制备登革病毒2-4型的prM和包膜蛋白的表达载体，其中包膜区域的氨基酸位置108(对应于SEQ ID NO.22至24的289aa)被从F置换为A，且包膜蛋白的C端区域(478-676aa或476-674aa)被置换为对应于登革病毒1型包膜蛋白的区域(SEQ ID NO.22的氨基酸478-676)。待由所述载体表达的蛋白为由SEQ ID NO.22-24表示的那些。病毒样颗粒以与实施例1相同的方式获得。

序列表

<110> VLP THERAPEUTICS, LLP

<120> 黄热病毒病毒样颗粒

<130> 6725231

<150> 62/048,897

<151> 2014-09-11

<160> 24

<170> PatentIn version 3.5

<210> 1

<211> 5498

<212> DNA

<213> 登革病毒

<220>

<221> misc_feature

<223> DENV1 prM E (Env F108A)

<220>

<221> CDS

<222> (1046)..(3073)

<400> 1

tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 60

cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 120

ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 180

accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 240

attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 300

tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 360

tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gacgcgtttg acattgatta 420

ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 480

ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 540

ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 600

cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 660

atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 720

cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 780

attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 840

cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 900

caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 960

cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gataactgca 1020

ggtcgacgat atcgcggccg ccacc atg tcg gtg acc atg ctc ctc atg ctg 1072

Met Ser Val Thr Met Leu Leu Met Leu

1 5

ctg ccc aca gcc ctg gcg ttc cat ctg acc acc cga ggg gga gag ccg 1120

Leu Pro Thr Ala Leu Ala Phe His Leu Thr Thr Arg Gly Gly Glu Pro

10 15 20 25

cac atg ata gtt agc aag cag gaa aga gga aaa tca ctt ttg ttt aag 1168

His Met Ile Val Ser Lys Gln Glu Arg Gly Lys Ser Leu Leu Phe Lys

30 35 40

acc tct gca ggt gtc aac atg tgc acc ctt att gca atg gat ttg gga 1216

Thr Ser Ala Gly Val Asn Met Cys Thr Leu Ile Ala Met Asp Leu Gly

45 50 55

gag tta tgt gag gac aca atg acc tac aaa tgc ccc cgg atc act gag 1264

Glu Leu Cys Glu Asp Thr Met Thr Tyr Lys Cys Pro Arg Ile Thr Glu

60 65 70

acg gaa cca gat gac gtt gac tgt tgg tgc aat gcc acg gag aca tgg 1312

Thr Glu Pro Asp Asp Val Asp Cys Trp Cys Asn Ala Thr Glu Thr Trp

75 80 85

gtg acc tat gga aca tgt tct caa act ggt gaa cac cga cga gac aaa 1360

Val Thr Tyr Gly Thr Cys Ser Gln Thr Gly Glu His Arg Arg Asp Lys

90 95 100 105

cgt tcc gtc gca ctg gca cca cac gta ggg ctt ggt cta gaa aca aga 1408

Arg Ser Val Ala Leu Ala Pro His Val Gly Leu Gly Leu Glu Thr Arg

110 115 120

acc gaa acg tgg atg tcc tct gaa ggc gct tgg aaa caa ata caa aaa 1456

Thr Glu Thr Trp Met Ser Ser Glu Gly Ala Trp Lys Gln Ile Gln Lys

125 130 135

gtg gag acc tgg gct ctg aga cac cca gga ttc acg gtg ata gcc ctt 1504

Val Glu Thr Trp Ala Leu Arg His Pro Gly Phe Thr Val Ile Ala Leu

140 145 150

ttt cta gca cat gcc ata gga aca tcc atc acc cag aaa ggg atc att 1552

Phe Leu Ala His Ala Ile Gly Thr Ser Ile Thr Gln Lys Gly Ile Ile

155 160 165

ttt att ttg ctg atg ctg gta act cca tcc atg gcc atg cgg tgc gtg 1600

Phe Ile Leu Leu Met Leu Val Thr Pro Ser Met Ala Met Arg Cys Val

170 175 180 185

gga ata ggc aac aga gac ttc gtg gaa gga ctg tca gga gct acg tgg 1648

Gly Ile Gly Asn Arg Asp Phe Val Glu Gly Leu Ser Gly Ala Thr Trp

190 195 200

gtg gat gtg gta ctg gag cat gga agt tgc gtc act acc atg gca aaa 1696

Val Asp Val Val Leu Glu His Gly Ser Cys Val Thr Thr Met Ala Lys

205 210 215

gac aaa cca aca ctg gac att gaa ctc ttg aag acg gag gtc aca aac 1744

Asp Lys Pro Thr Leu Asp Ile Glu Leu Leu Lys Thr Glu Val Thr Asn

220 225 230

cct gcc gtc ctg cgc aaa ctg tgc att gaa gct aaa ata tca aac acc 1792

Pro Ala Val Leu Arg Lys Leu Cys Ile Glu Ala Lys Ile Ser Asn Thr

235 240 245

acc acc gat tcg aga tgt cca aca caa gga gaa gcc acg ctg gtg gaa 1840

Thr Thr Asp Ser Arg Cys Pro Thr Gln Gly Glu Ala Thr Leu Val Glu

250 255 260 265

gaa cag gac acg aac ttt gtg tgt cga cga acg ttc gtg gac aga ggc 1888

Glu Gln Asp Thr Asn Phe Val Cys Arg Arg Thr Phe Val Asp Arg Gly

270 275 280

tgg ggc aat ggt tgt ggg cta gcc gga aaa ggt agc tta ata acg tgt 1936

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

285 290 295

gct aag ttt aag tgt gtg aca aaa ctg gaa gga aag ata gtc caa tat 1984

Ala Lys Phe Lys Cys Val Thr Lys Leu Glu Gly Lys Ile Val Gln Tyr

300 305 310

gaa aac tta aaa tat tca gtg ata gtc acc gta cac act gga gac cag 2032

Glu Asn Leu Lys Tyr Ser Val Ile Val Thr Val His Thr Gly Asp Gln

315 320 325

cac caa gtt gga aat gag acc aca gaa cat gga aca act gca acc ata 2080

His Gln Val Gly Asn Glu Thr Thr Glu His Gly Thr Thr Ala Thr Ile

330 335 340 345

aca cct caa gct ccc acg tcg gaa ata cag ctg aca gac tac gga gct 2128

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

350 355 360

cta aca ttg gat tgt tca cct aga aca ggg cta gac ttt aat gag atg 2176

Leu Thr Leu Asp Cys Ser Pro Arg Thr Gly Leu Asp Phe Asn Glu Met

365 370 375

gtg ttg ttg aca atg gaa aaa aaa tca tgg ctc gtc cac aaa caa tgg 2224

Val Leu Leu Thr Met Glu Lys Lys Ser Trp Leu Val His Lys Gln Trp

380 385 390

ttt cta gac tta cca ctg cct tgg acc tcg ggg gct tca aca tcc caa 2272

Phe Leu Asp Leu Pro Leu Pro Trp Thr Ser Gly Ala Ser Thr Ser Gln

395 400 405

gag act tgg aat aga caa gac ttg ctg gtc aca ttt aag aca gct cat 2320

Glu Thr Trp Asn Arg Gln Asp Leu Leu Val Thr Phe Lys Thr Ala His

410 415 420 425

gca aaa aag cag gaa gta gtc gta cta gga tca caa gaa gga gca atg 2368

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

430 435 440

cac act gcg ttg act gga gcg aca gaa atc caa acg tct gga acg aca 2416

His Thr Ala Leu Thr Gly Ala Thr Glu Ile Gln Thr Ser Gly Thr Thr

445 450 455

aca att ttt gca gga cac ctg aaa tgc aga cta aaa atg gat aaa ctg 2464

Thr Ile Phe Ala Gly His Leu Lys Cys Arg Leu Lys Met Asp Lys Leu

460 465 470

act tta aaa ggg atg tca tat gta atg tgc aca ggg tca ttc aag tta 2512

Thr Leu Lys Gly Met Ser Tyr Val Met Cys Thr Gly Ser Phe Lys Leu

475 480 485

gag aag gaa gtg gct gag acc cag cat gga act gtt cta gtg cag gtt 2560

Glu Lys Glu Val Ala Glu Thr Gln His Gly Thr Val Leu Val Gln Val

490 495 500 505

aaa tac gaa gga aca gat gca cca tgc aag atc ccc ttc tcg tcc caa 2608

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

510 515 520

gat gag aag gga gta acc cag aat ggg aga ttg ata aca gcc aac ccc 2656

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

525 530 535

ata gtc act gac aaa gaa aaa cca gtc aac att gaa gcg gag cca cct 2704

Ile Val Thr Asp Lys Glu Lys Pro Val Asn Ile Glu Ala Glu Pro Pro

540 545 550

ttt ggt gag agc tac att gtg gta gga gca ggt gaa aaa gct ttg aaa 2752

Phe Gly Glu Ser Tyr Ile Val Val Gly Ala Gly Glu Lys Ala Leu Lys

555 560 565

cta agc tgg ttc aag aag gga agc agt ata ggg aaa atg ttt gaa gca 2800

Leu Ser Trp Phe Lys Lys Gly Ser Ser Ile Gly Lys Met Phe Glu Ala

570 575 580 585

act gcc cgt gga gca cga agg atg gcc atc ctg gga gac act gca tgg 2848

Thr Ala Arg Gly Ala Arg Arg Met Ala Ile Leu Gly Asp Thr Ala Trp

590 595 600

gac ttc ggt tct ata gga ggg gtg ttc acg tct gtg gga aaa ctg ata 2896

Asp Phe Gly Ser Ile Gly Gly Val Phe Thr Ser Val Gly Lys Leu Ile

605 610 615

cac cag att ttt ggg act gcg tat gga gtt ttg ttc agc ggt gtt tct 2944

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

620 625 630

tgg acc atg aag ata gga ata ggg att ctg ctg aca tgg cta gga tta 2992

Trp Thr Met Lys Ile Gly Ile Gly Ile Leu Leu Thr Trp Leu Gly Leu

635 640 645

aac tca agg agc acg tcc ctt tca atg acg tgt atc gca gtt ggc atg 3040

Asn Ser Arg Ser Thr Ser Leu Ser Met Thr Cys Ile Ala Val Gly Met

650 655 660 665

gtc acg ctg tac cta gga gtc atg gtt cag gcg taataaagat ctgctgtgcc 3093

Val Thr Leu Tyr Leu Gly Val Met Val Gln Ala

670 675

ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 3153

tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 3213

gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 3273

caatagcagg catgctgggg atgcggtggg ctctatggct cgagcatggt catagctgtt 3333

tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa 3393

gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact 3453

gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 3513

ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg 3573

ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 3633

cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag 3693

gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 3753

tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 3813

ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 3873

atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 3933

gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 3993

tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 4053

cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 4113

cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt 4173

tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 4233

cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 4293

cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 4353

gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 4413

gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg 4473

gtctgacagt tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg 4533

attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag 4593

gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc 4653

aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg 4713

agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc 4773

aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat 4833

tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac 4893

aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga 4953

atcaggatat tcttctaata cctggaatgc tgttttccca gggatcgcag tggtgagtaa 5013

ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt 5073

cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg 5133

tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga 5193

ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt 5253

taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcatactct tcctttttca 5313

atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 5373

ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 5433

ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt 5493

tcgtc 5498

<210> 2

<211> 676

<212> PRT

<213> 登革病毒

<400> 2

Met Ser Val Thr Met Leu Leu Met Leu Leu Pro Thr Ala Leu Ala Phe

1 5 10 15

His Leu Thr Thr Arg Gly Gly Glu Pro His Met Ile Val Ser Lys Gln

20 25 30

Glu Arg Gly Lys Ser Leu Leu Phe Lys Thr Ser Ala Gly Val Asn Met

35 40 45

Cys Thr Leu Ile Ala Met Asp Leu Gly Glu Leu Cys Glu Asp Thr Met

50 55 60

Thr Tyr Lys Cys Pro Arg Ile Thr Glu Thr Glu Pro Asp Asp Val Asp

65 70 75 80

Cys Trp Cys Asn Ala Thr Glu Thr Trp Val Thr Tyr Gly Thr Cys Ser

85 90 95

Gln Thr Gly Glu His Arg Arg Asp Lys Arg Ser Val Ala Leu Ala Pro

100 105 110

His Val Gly Leu Gly Leu Glu Thr Arg Thr Glu Thr Trp Met Ser Ser

115 120 125

Glu Gly Ala Trp Lys Gln Ile Gln Lys Val Glu Thr Trp Ala Leu Arg

130 135 140

His Pro Gly Phe Thr Val Ile Ala Leu Phe Leu Ala His Ala Ile Gly

145 150 155 160

Thr Ser Ile Thr Gln Lys Gly Ile Ile Phe Ile Leu Leu Met Leu Val

165 170 175

Thr Pro Ser Met Ala Met Arg Cys Val Gly Ile Gly Asn Arg Asp Phe

180 185 190

Val Glu Gly Leu Ser Gly Ala Thr Trp Val Asp Val Val Leu Glu His

195 200 205

Gly Ser Cys Val Thr Thr Met Ala Lys Asp Lys Pro Thr Leu Asp Ile

210 215 220

Glu Leu Leu Lys Thr Glu Val Thr Asn Pro Ala Val Leu Arg Lys Leu

225 230 235 240

Cys Ile Glu Ala Lys Ile Ser Asn Thr Thr Thr Asp Ser Arg Cys Pro

245 250 255

Thr Gln Gly Glu Ala Thr Leu Val Glu Glu Gln Asp Thr Asn Phe Val

260 265 270

Cys Arg Arg Thr Phe Val Asp Arg Gly Trp Gly Asn Gly Cys Gly Leu

275 280 285

Ala Gly Lys Gly Ser Leu Ile Thr Cys Ala Lys Phe Lys Cys Val Thr

290 295 300

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

305 310 315 320

Ile Val Thr Val His Thr Gly Asp Gln His Gln Val Gly Asn Glu Thr

325 330 335

Thr Glu His Gly Thr Thr Ala Thr Ile Thr Pro Gln Ala Pro Thr Ser

340 345 350

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

355 360 365

Arg Thr Gly Leu Asp Phe Asn Glu Met Val Leu Leu Thr Met Glu Lys

370 375 380

Lys Ser Trp Leu Val His Lys Gln Trp Phe Leu Asp Leu Pro Leu Pro

385 390 395 400

Trp Thr Ser Gly Ala Ser Thr Ser Gln Glu Thr Trp Asn Arg Gln Asp

405 410 415

Leu Leu Val Thr Phe Lys Thr Ala His Ala Lys Lys Gln Glu Val Val

420 425 430

Val Leu Gly Ser Gln Glu Gly Ala Met His Thr Ala Leu Thr Gly Ala

435 440 445

Thr Glu Ile Gln Thr Ser Gly Thr Thr Thr Ile Phe Ala Gly His Leu

450 455 460

Lys Cys Arg Leu Lys Met Asp Lys Leu Thr Leu Lys Gly Met Ser Tyr

465 470 475 480

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

485 490 495

Gln His Gly Thr Val Leu Val Gln Val Lys Tyr Glu Gly Thr Asp Ala

500 505 510

Pro Cys Lys Ile Pro Phe Ser Ser Gln Asp Glu Lys Gly Val Thr Gln

515 520 525

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

530 535 540

Pro Val Asn Ile Glu Ala Glu Pro Pro Phe Gly Glu Ser Tyr Ile Val

545 550 555 560

Val Gly Ala Gly Glu Lys Ala Leu Lys Leu Ser Trp Phe Lys Lys Gly

565 570 575

Ser Ser Ile Gly Lys Met Phe Glu Ala Thr Ala Arg Gly Ala Arg Arg

580 585 590

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

595 600 605

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

610 615 620

Tyr Gly Val Leu Phe Ser Gly Val Ser Trp Thr Met Lys Ile Gly Ile

625 630 635 640

Gly Ile Leu Leu Thr Trp Leu Gly Leu Asn Ser Arg Ser Thr Ser Leu

645 650 655

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

660 665 670

Met Val Gln Ala

675

<210> 3

<211> 5498

<212> DNA

<213> 登革病毒

<220>

<221> misc_feature

<223> DENV1 prM E (Env K203N)

<220>

<221> CDS

<222> (1046)..(3073)

<400> 3

tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 60

cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 120

ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 180

accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 240

attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 300

tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 360

tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gacgcgtttg acattgatta 420

ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 480

ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 540

ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 600

cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 660

atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 720

cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 780

attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 840

cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 900

caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 960

cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gataactgca 1020

ggtcgacgat atcgcggccg ccacc atg tcg gtg acc atg ctc ctc atg ctg 1072

Met Ser Val Thr Met Leu Leu Met Leu

1 5

ctg ccc aca gcc ctg gcg ttc cat ctg acc acc cga ggg gga gag ccg 1120

Leu Pro Thr Ala Leu Ala Phe His Leu Thr Thr Arg Gly Gly Glu Pro

10 15 20 25

cac atg ata gtt agc aag cag gaa aga gga aaa tca ctt ttg ttt aag 1168

His Met Ile Val Ser Lys Gln Glu Arg Gly Lys Ser Leu Leu Phe Lys

30 35 40

acc tct gca ggt gtc aac atg tgc acc ctt att gca atg gat ttg gga 1216

Thr Ser Ala Gly Val Asn Met Cys Thr Leu Ile Ala Met Asp Leu Gly

45 50 55

gag tta tgt gag gac aca atg acc tac aaa tgc ccc cgg atc act gag 1264

Glu Leu Cys Glu Asp Thr Met Thr Tyr Lys Cys Pro Arg Ile Thr Glu

60 65 70

acg gaa cca gat gac gtt gac tgt tgg tgc aat gcc acg gag aca tgg 1312

Thr Glu Pro Asp Asp Val Asp Cys Trp Cys Asn Ala Thr Glu Thr Trp

75 80 85

gtg acc tat gga aca tgt tct caa act ggt gaa cac cga cga gac aaa 1360

Val Thr Tyr Gly Thr Cys Ser Gln Thr Gly Glu His Arg Arg Asp Lys

90 95 100 105

cgt tcc gtc gca ctg gca cca cac gta ggg ctt ggt cta gaa aca aga 1408

Arg Ser Val Ala Leu Ala Pro His Val Gly Leu Gly Leu Glu Thr Arg

110 115 120

acc gaa acg tgg atg tcc tct gaa ggc gct tgg aaa caa ata caa aaa 1456

Thr Glu Thr Trp Met Ser Ser Glu Gly Ala Trp Lys Gln Ile Gln Lys

125 130 135

gtg gag acc tgg gct ctg aga cac cca gga ttc acg gtg ata gcc ctt 1504

Val Glu Thr Trp Ala Leu Arg His Pro Gly Phe Thr Val Ile Ala Leu

140 145 150

ttt cta gca cat gcc ata gga aca tcc atc acc cag aaa ggg atc att 1552

Phe Leu Ala His Ala Ile Gly Thr Ser Ile Thr Gln Lys Gly Ile Ile

155 160 165

ttt att ttg ctg atg ctg gta act cca tcc atg gcc atg cgg tgc gtg 1600

Phe Ile Leu Leu Met Leu Val Thr Pro Ser Met Ala Met Arg Cys Val

170 175 180 185

gga ata ggc aac aga gac ttc gtg gaa gga ctg tca gga gct acg tgg 1648

Gly Ile Gly Asn Arg Asp Phe Val Glu Gly Leu Ser Gly Ala Thr Trp

190 195 200

gtg gat gtg gta ctg gag cat gga agt tgc gtc act acc atg gca aaa 1696

Val Asp Val Val Leu Glu His Gly Ser Cys Val Thr Thr Met Ala Lys

205 210 215

gac aaa cca aca ctg gac att gaa ctc ttg aag acg gag gtc aca aac 1744

Asp Lys Pro Thr Leu Asp Ile Glu Leu Leu Lys Thr Glu Val Thr Asn

220 225 230

cct gcc gtc ctg cgc aaa ctg tgc att gaa gct aaa ata tca aac acc 1792

Pro Ala Val Leu Arg Lys Leu Cys Ile Glu Ala Lys Ile Ser Asn Thr

235 240 245

acc acc gat tcg aga tgt cca aca caa gga gaa gcc acg ctg gtg gaa 1840

Thr Thr Asp Ser Arg Cys Pro Thr Gln Gly Glu Ala Thr Leu Val Glu

250 255 260 265

gaa cag gac acg aac ttt gtg tgt cga cga acg ttc gtg gac aga ggc 1888

Glu Gln Asp Thr Asn Phe Val Cys Arg Arg Thr Phe Val Asp Arg Gly

270 275 280

tgg ggc aat ggt tgt ggg cta ttc gga aaa ggt agc tta ata acg tgt 1936

Trp Gly Asn Gly Cys Gly Leu Phe Gly Lys Gly Ser Leu Ile Thr Cys

285 290 295

gct aag ttt aag tgt gtg aca aaa ctg gaa gga aag ata gtc caa tat 1984

Ala Lys Phe Lys Cys Val Thr Lys Leu Glu Gly Lys Ile Val Gln Tyr

300 305 310

gaa aac tta aaa tat tca gtg ata gtc acc gta cac act gga gac cag 2032

Glu Asn Leu Lys Tyr Ser Val Ile Val Thr Val His Thr Gly Asp Gln

315 320 325

cac caa gtt gga aat gag acc aca gaa cat gga aca act gca acc ata 2080

His Gln Val Gly Asn Glu Thr Thr Glu His Gly Thr Thr Ala Thr Ile

330 335 340 345

aca cct caa gct ccc acg tcg gaa ata cag ctg aca gac tac gga gct 2128

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

350 355 360

cta aca ttg gat tgt tca cct aga aca ggg cta gac ttt aat gag atg 2176

Leu Thr Leu Asp Cys Ser Pro Arg Thr Gly Leu Asp Phe Asn Glu Met

365 370 375

gtg ttg ttg aca atg gaa aat aaa tca tgg ctc gtc cac aaa caa tgg 2224

Val Leu Leu Thr Met Glu Asn Lys Ser Trp Leu Val His Lys Gln Trp

380 385 390

ttt cta gac tta cca ctg cct tgg acc tcg ggg gct tca aca tcc caa 2272

Phe Leu Asp Leu Pro Leu Pro Trp Thr Ser Gly Ala Ser Thr Ser Gln

395 400 405

gag act tgg aat aga caa gac ttg ctg gtc aca ttt aag aca gct cat 2320

Glu Thr Trp Asn Arg Gln Asp Leu Leu Val Thr Phe Lys Thr Ala His

410 415 420 425

gca aaa aag cag gaa gta gtc gta cta gga tca caa gaa gga gca atg 2368

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

430 435 440

cac act gcg ttg act gga gcg aca gaa atc caa acg tct gga acg aca 2416

His Thr Ala Leu Thr Gly Ala Thr Glu Ile Gln Thr Ser Gly Thr Thr

445 450 455

aca att ttt gca gga cac ctg aaa tgc aga cta aaa atg gat aaa ctg 2464

Thr Ile Phe Ala Gly His Leu Lys Cys Arg Leu Lys Met Asp Lys Leu

460 465 470

act tta aaa ggg atg tca tat gta atg tgc aca ggg tca ttc aag tta 2512

Thr Leu Lys Gly Met Ser Tyr Val Met Cys Thr Gly Ser Phe Lys Leu

475 480 485

gag aag gaa gtg gct gag acc cag cat gga act gtt cta gtg cag gtt 2560

Glu Lys Glu Val Ala Glu Thr Gln His Gly Thr Val Leu Val Gln Val

490 495 500 505

aaa tac gaa gga aca gat gca cca tgc aag atc ccc ttc tcg tcc caa 2608

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

510 515 520

gat gag aag gga gta acc cag aat ggg aga ttg ata aca gcc aac ccc 2656

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

525 530 535

ata gtc act gac aaa gaa aaa cca gtc aac att gaa gcg gag cca cct 2704

Ile Val Thr Asp Lys Glu Lys Pro Val Asn Ile Glu Ala Glu Pro Pro

540 545 550

ttt ggt gag agc tac att gtg gta gga gca ggt gaa aaa gct ttg aaa 2752

Phe Gly Glu Ser Tyr Ile Val Val Gly Ala Gly Glu Lys Ala Leu Lys

555 560 565

cta agc tgg ttc aag aag gga agc agt ata ggg aaa atg ttt gaa gca 2800

Leu Ser Trp Phe Lys Lys Gly Ser Ser Ile Gly Lys Met Phe Glu Ala

570 575 580 585

act gcc cgt gga gca cga agg atg gcc atc ctg gga gac act gca tgg 2848

Thr Ala Arg Gly Ala Arg Arg Met Ala Ile Leu Gly Asp Thr Ala Trp

590 595 600

gac ttc ggt tct ata gga ggg gtg ttc acg tct gtg gga aaa ctg ata 2896

Asp Phe Gly Ser Ile Gly Gly Val Phe Thr Ser Val Gly Lys Leu Ile

605 610 615

cac cag att ttt ggg act gcg tat gga gtt ttg ttc agc ggt gtt tct 2944

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

620 625 630

tgg acc atg aag ata gga ata ggg att ctg ctg aca tgg cta gga tta 2992

Trp Thr Met Lys Ile Gly Ile Gly Ile Leu Leu Thr Trp Leu Gly Leu

635 640 645

aac tca agg agc acg tcc ctt tca atg acg tgt atc gca gtt ggc atg 3040

Asn Ser Arg Ser Thr Ser Leu Ser Met Thr Cys Ile Ala Val Gly Met

650 655 660 665

gtc acg ctg tac cta gga gtc atg gtt cag gcg taataaagat ctgctgtgcc 3093

Val Thr Leu Tyr Leu Gly Val Met Val Gln Ala

670 675

ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 3153

tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 3213

gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 3273

caatagcagg catgctgggg atgcggtggg ctctatggct cgagcatggt catagctgtt 3333

tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa 3393

gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact 3453

gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 3513

ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg 3573

ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 3633

cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag 3693

gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 3753

tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 3813

ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 3873

atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 3933

gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 3993

tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 4053

cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 4113

cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt 4173

tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 4233

cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 4293

cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 4353

gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 4413

gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg 4473

gtctgacagt tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg 4533

attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag 4593

gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc 4653

aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg 4713

agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc 4773

aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat 4833

tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac 4893

aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga 4953

atcaggatat tcttctaata cctggaatgc tgttttccca gggatcgcag tggtgagtaa 5013

ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt 5073

cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg 5133

tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga 5193

ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt 5253

taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcatactct tcctttttca 5313

atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 5373

ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 5433

ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt 5493

tcgtc 5498

<210> 4

<211> 676

<212> PRT

<213> 登革病毒

<400> 4

Met Ser Val Thr Met Leu Leu Met Leu Leu Pro Thr Ala Leu Ala Phe

1 5 10 15

His Leu Thr Thr Arg Gly Gly Glu Pro His Met Ile Val Ser Lys Gln

20 25 30

Glu Arg Gly Lys Ser Leu Leu Phe Lys Thr Ser Ala Gly Val Asn Met

35 40 45

Cys Thr Leu Ile Ala Met Asp Leu Gly Glu Leu Cys Glu Asp Thr Met

50 55 60

Thr Tyr Lys Cys Pro Arg Ile Thr Glu Thr Glu Pro Asp Asp Val Asp

65 70 75 80

Cys Trp Cys Asn Ala Thr Glu Thr Trp Val Thr Tyr Gly Thr Cys Ser

85 90 95

Gln Thr Gly Glu His Arg Arg Asp Lys Arg Ser Val Ala Leu Ala Pro

100 105 110

His Val Gly Leu Gly Leu Glu Thr Arg Thr Glu Thr Trp Met Ser Ser

115 120 125

Glu Gly Ala Trp Lys Gln Ile Gln Lys Val Glu Thr Trp Ala Leu Arg

130 135 140

His Pro Gly Phe Thr Val Ile Ala Leu Phe Leu Ala His Ala Ile Gly

145 150 155 160

Thr Ser Ile Thr Gln Lys Gly Ile Ile Phe Ile Leu Leu Met Leu Val

165 170 175

Thr Pro Ser Met Ala Met Arg Cys Val Gly Ile Gly Asn Arg Asp Phe

180 185 190

Val Glu Gly Leu Ser Gly Ala Thr Trp Val Asp Val Val Leu Glu His

195 200 205

Gly Ser Cys Val Thr Thr Met Ala Lys Asp Lys Pro Thr Leu Asp Ile

210 215 220

Glu Leu Leu Lys Thr Glu Val Thr Asn Pro Ala Val Leu Arg Lys Leu

225 230 235 240

Cys Ile Glu Ala Lys Ile Ser Asn Thr Thr Thr Asp Ser Arg Cys Pro

245 250 255

Thr Gln Gly Glu Ala Thr Leu Val Glu Glu Gln Asp Thr Asn Phe Val

260 265 270

Cys Arg Arg Thr Phe Val Asp Arg Gly Trp Gly Asn Gly Cys Gly Leu

275 280 285

Phe Gly Lys Gly Ser Leu Ile Thr Cys Ala Lys Phe Lys Cys Val Thr

290 295 300

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

305 310 315 320

Ile Val Thr Val His Thr Gly Asp Gln His Gln Val Gly Asn Glu Thr

325 330 335

Thr Glu His Gly Thr Thr Ala Thr Ile Thr Pro Gln Ala Pro Thr Ser

340 345 350

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

355 360 365

Arg Thr Gly Leu Asp Phe Asn Glu Met Val Leu Leu Thr Met Glu Asn

370 375 380

Lys Ser Trp Leu Val His Lys Gln Trp Phe Leu Asp Leu Pro Leu Pro

385 390 395 400

Trp Thr Ser Gly Ala Ser Thr Ser Gln Glu Thr Trp Asn Arg Gln Asp

405 410 415

Leu Leu Val Thr Phe Lys Thr Ala His Ala Lys Lys Gln Glu Val Val

420 425 430

Val Leu Gly Ser Gln Glu Gly Ala Met His Thr Ala Leu Thr Gly Ala

435 440 445

Thr Glu Ile Gln Thr Ser Gly Thr Thr Thr Ile Phe Ala Gly His Leu

450 455 460

Lys Cys Arg Leu Lys Met Asp Lys Leu Thr Leu Lys Gly Met Ser Tyr

465 470 475 480

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

485 490 495

Gln His Gly Thr Val Leu Val Gln Val Lys Tyr Glu Gly Thr Asp Ala

500 505 510

Pro Cys Lys Ile Pro Phe Ser Ser Gln Asp Glu Lys Gly Val Thr Gln

515 520 525

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

530 535 540

Pro Val Asn Ile Glu Ala Glu Pro Pro Phe Gly Glu Ser Tyr Ile Val

545 550 555 560

Val Gly Ala Gly Glu Lys Ala Leu Lys Leu Ser Trp Phe Lys Lys Gly

565 570 575

Ser Ser Ile Gly Lys Met Phe Glu Ala Thr Ala Arg Gly Ala Arg Arg

580 585 590

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

595 600 605

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

610 615 620

Tyr Gly Val Leu Phe Ser Gly Val Ser Trp Thr Met Lys Ile Gly Ile

625 630 635 640

Gly Ile Leu Leu Thr Trp Leu Gly Leu Asn Ser Arg Ser Thr Ser Leu

645 650 655

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

660 665 670

Met Val Gln Ala

675

<210> 5

<211> 3770

<212> DNA

<213> 登革病毒

<220>

<221> misc_feature

<223> DENV1衣壳蛋白K85A_K86A突变体

<220>

<221> CDS

<222> (1046)..(1345)

<400> 5

tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 60

cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 120

ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 180

accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 240

attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 300

tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 360

tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gacgcgtttg acattgatta 420

ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 480

ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 540

ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 600

cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 660

atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 720

cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 780

attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 840

cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 900

caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 960

cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gataactgca 1020

ggtcgacgat atcgcggccg ccacc atg aac aac caa cgg aaa aag acg ggt 1072

Met Asn Asn Gln Arg Lys Lys Thr Gly

1 5

cga ccg tct ttc aat atg ctg aaa cgc gcg aga aac cgc gtg tca act 1120

Arg Pro Ser Phe Asn Met Leu Lys Arg Ala Arg Asn Arg Val Ser Thr

10 15 20 25

gtt tca cag ttg gcg aag aga ttc tca aaa gga ttg ctt tca ggc caa 1168

Val Ser Gln Leu Ala Lys Arg Phe Ser Lys Gly Leu Leu Ser Gly Gln

30 35 40

gga ccc atg aaa ttg gtg atg gct ttt ata gca ttc cta aga ttt cta 1216

Gly Pro Met Lys Leu Val Met Ala Phe Ile Ala Phe Leu Arg Phe Leu

45 50 55

gcc ata cct cca aca gca gga att ttg gct aga tgg ggc tca ttc aag 1264

Ala Ile Pro Pro Thr Ala Gly Ile Leu Ala Arg Trp Gly Ser Phe Lys

60 65 70

aag aat gga gcg atc aaa gtg tta cgg ggt ttc gcg gca gaa atc tca 1312

Lys Asn Gly Ala Ile Lys Val Leu Arg Gly Phe Ala Ala Glu Ile Ser

75 80 85

aac atg ttg aac ata atg aac agg agg aaa aga taataaagat ctgctgtgcc 1365

Asn Met Leu Asn Ile Met Asn Arg Arg Lys Arg

90 95 100

ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 1425

tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 1485

gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 1545

caatagcagg catgctgggg atgcggtggg ctctatggct cgagcatggt catagctgtt 1605

tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa 1665

gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact 1725

gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 1785

ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg 1845

ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 1905

cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag 1965

gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 2025

tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 2085

ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 2145

atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 2205

gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 2265

tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 2325

cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 2385

cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt 2445

tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 2505

cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 2565

cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 2625

gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 2685

gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg 2745

gtctgacagt tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg 2805

attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag 2865

gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc 2925

aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg 2985

agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc 3045

aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat 3105

tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac 3165

aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga 3225

atcaggatat tcttctaata cctggaatgc tgttttccca gggatcgcag tggtgagtaa 3285

ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt 3345

cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg 3405

tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga 3465

ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt 3525

taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcatactct tcctttttca 3585

atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 3645

ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 3705

ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt 3765

tcgtc 3770

<210> 6

<211> 100

<212> PRT

<213> 登革病毒

<400> 6

Met Asn Asn Gln Arg Lys Lys Thr Gly Arg Pro Ser Phe Asn Met Leu

1 5 10 15

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

20 25 30

Phe Ser Lys Gly Leu Leu Ser Gly Gln Gly Pro Met Lys Leu Val Met

35 40 45

Ala Phe Ile Ala Phe Leu Arg Phe Leu Ala Ile Pro Pro Thr Ala Gly

50 55 60

Ile Leu Ala Arg Trp Gly Ser Phe Lys Lys Asn Gly Ala Ile Lys Val

65 70 75 80

Leu Arg Gly Phe Ala Ala Glu Ile Ser Asn Met Leu Asn Ile Met Asn

85 90 95

Arg Arg Lys Arg

100

<210> 7

<211> 6679

<212> DNA

<213> 登革病毒

<220>

<221> misc_feature

<223> 嵌合体DENV2 prM E_嵌合DENV1 E

<220>

<221> CDS

<222> (906)..(2933)

<220>

<221> misc_feature

<222> (906)..(2336)

<223> DENV2 prM E区域

<220>

<221> misc_feature

<222> (2337)..(2933)

<223> DENV1 E区域

<400> 7

gaattcccat tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt 60

ccaacattac cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg 120

gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc 180

ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc 240

atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact 300

gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat 360

gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact 420

tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac 480

atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac 540

gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac 600

tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga 660

gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat 720

agaagacacc gggaccgatc cagcctccgt taacggtgga gggcagtgta gtctgagcag 780

tactcgttgc tgccgcgcgc gccaccagac ataatagctg acagactaac agactgttcc 840

tttccatggg tcttttctgc agtcaccgtc gtcgacacgt gtgatcagat atcgcggccg 900

ccacc atg act gca ggc atg atc atc atg ctg att cca aca gtg atg gcg 950

Met Thr Ala Gly Met Ile Ile Met Leu Ile Pro Thr Val Met Ala

1 5 10 15

ttt cat ctg acc aca cgc aac gga gaa cca cac atg atc gtc agt aga 998

Phe His Leu Thr Thr Arg Asn Gly Glu Pro His Met Ile Val Ser Arg

20 25 30

caa gaa aaa ggg aaa agc ctt ctg ttt aag aca aag gac ggc acg aac 1046

Gln Glu Lys Gly Lys Ser Leu Leu Phe Lys Thr Lys Asp Gly Thr Asn

35 40 45

atg tgt acc ctc atg gcc atg gac ctt ggt gag ttg tgt gaa gac aca 1094

Met Cys Thr Leu Met Ala Met Asp Leu Gly Glu Leu Cys Glu Asp Thr

50 55 60

atc acg tat aaa tgt ccc ttt ctc aag cag aac gaa cca gaa gac ata 1142

Ile Thr Tyr Lys Cys Pro Phe Leu Lys Gln Asn Glu Pro Glu Asp Ile

65 70 75

gat tgt tgg tgc aac tcc acg tcc aca tgg gta act tat ggg aca tgt 1190

Asp Cys Trp Cys Asn Ser Thr Ser Thr Trp Val Thr Tyr Gly Thr Cys

80 85 90 95

acc acc aca gga gag cac aga aga gaa aaa aga tca gtg gcg ctt gtt 1238

Thr Thr Thr Gly Glu His Arg Arg Glu Lys Arg Ser Val Ala Leu Val

100 105 110

cca cac gtg gga atg gga ttg gag aca cga act gaa aca tgg atg tca 1286

Pro His Val Gly Met Gly Leu Glu Thr Arg Thr Glu Thr Trp Met Ser

115 120 125

tca gaa ggg gcc tgg aaa cat gcc cag aga att gaa act tgg att ctg 1334

Ser Glu Gly Ala Trp Lys His Ala Gln Arg Ile Glu Thr Trp Ile Leu

130 135 140

aga cat cca ggc ttt acc ata atg gcc gca atc ctg gca tac acc ata 1382

Arg His Pro Gly Phe Thr Ile Met Ala Ala Ile Leu Ala Tyr Thr Ile

145 150 155

gga acg acg cat ttc caa aga gtc ctg ata ttc atc cta ctg aca gcc 1430

Gly Thr Thr His Phe Gln Arg Val Leu Ile Phe Ile Leu Leu Thr Ala

160 165 170 175

atc gct cct tca atg aca atg cgc tgc ata gga ata tca aat agg gac 1478

Ile Ala Pro Ser Met Thr Met Arg Cys Ile Gly Ile Ser Asn Arg Asp

180 185 190

ttt gtg gaa gga gtg tca gga ggg agt tgg gtt gac ata gtt tta gaa 1526

Phe Val Glu Gly Val Ser Gly Gly Ser Trp Val Asp Ile Val Leu Glu

195 200 205

cat gga agt tgt gtg acg acg atg gca aaa aat aaa cca aca ctg gac 1574

His Gly Ser Cys Val Thr Thr Met Ala Lys Asn Lys Pro Thr Leu Asp

210 215 220

ttt gaa ctg ata aaa aca gaa gcc aaa caa ccc gcc acc tta agg aag 1622

Phe Glu Leu Ile Lys Thr Glu Ala Lys Gln Pro Ala Thr Leu Arg Lys

225 230 235

tac tgt ata gag gct aaa ctg acc aac acg aca aca gac tcg cgc tgc 1670

Tyr Cys Ile Glu Ala Lys Leu Thr Asn Thr Thr Thr Asp Ser Arg Cys

240 245 250 255

cca aca caa ggg gaa ccc acc ctg aat gaa gag cag gac aaa agg ttt 1718

Pro Thr Gln Gly Glu Pro Thr Leu Asn Glu Glu Gln Asp Lys Arg Phe

260 265 270

gtc tgc aaa cat tcc atg gta gac aga gga tgg gga aat gga tgt gga 1766

Val Cys Lys His Ser Met Val Asp Arg Gly Trp Gly Asn Gly Cys Gly

275 280 285

tta ttt gga aaa gga ggc atc gtg acc tgt gcc atg ttc aca tgc aaa 1814

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

290 295 300

aag aac atg gag gga aaa att gtg cag cca gaa aac ctg gaa tac act 1862

Lys Asn Met Glu Gly Lys Ile Val Gln Pro Glu Asn Leu Glu Tyr Thr

305 310 315

gtc gtt ata aca cct cat tca ggg gaa gaa cat gca gtc gga aat gac 1910

Val Val Ile Thr Pro His Ser Gly Glu Glu His Ala Val Gly Asn Asp

320 325 330 335

aca gga aaa cat ggt aaa gaa gtc aag ata aca cca cag agc tcc atc 1958

Thr Gly Lys His Gly Lys Glu Val Lys Ile Thr Pro Gln Ser Ser Ile

340 345 350

aca gag gcg gaa ctg aca ggc tat ggc act gtt acg atg gag tgc tct 2006

Thr Glu Ala Glu Leu Thr Gly Tyr Gly Thr Val Thr Met Glu Cys Ser

355 360 365

cca aga acg ggc ctc gac ttc aat gag atg gtg ttg ctg caa atg aaa 2054

Pro Arg Thr Gly Leu Asp Phe Asn Glu Met Val Leu Leu Gln Met Lys

370 375 380

gac aaa gct tgg ctg gtg cac aga caa tgg ttc cta gac cta ccg ttg 2102

Asp Lys Ala Trp Leu Val His Arg Gln Trp Phe Leu Asp Leu Pro Leu

385 390 395

cca tgg ctg ccc gga gca gac aca caa gga tca aat tgg ata cag aaa 2150

Pro Trp Leu Pro Gly Ala Asp Thr Gln Gly Ser Asn Trp Ile Gln Lys

400 405 410 415

gag aca ctg gtc acc ttc aaa aat ccc cat gcg aaa aaa cag gat gtt 2198

Glu Thr Leu Val Thr Phe Lys Asn Pro His Ala Lys Lys Gln Asp Val

420 425 430

gtt gtc tta ggc tcc caa gag ggg gcc atg cat aca gca ctc aca ggg 2246

Val Val Leu Gly Ser Gln Glu Gly Ala Met His Thr Ala Leu Thr Gly

435 440 445

gct acg gaa atc cag atg tca tca gga aac ctg ctg ttc aca gga cat 2294

Ala Thr Glu Ile Gln Met Ser Ser Gly Asn Leu Leu Phe Thr Gly His

450 455 460

ctt aag tgc agg ctg aga atg gac aaa tta caa ctt aaa ggg atg tca 2342

Leu Lys Cys Arg Leu Arg Met Asp Lys Leu Gln Leu Lys Gly Met Ser

465 470 475

tat gta atg tgc aca ggg tca ttc aag tta gag aag gaa gtg gct gag 2390

Tyr Val Met Cys Thr Gly Ser Phe Lys Leu Glu Lys Glu Val Ala Glu

480 485 490 495

acc cag cat gga act gtt cta gtg cag gtt aaa tac gaa gga aca gat 2438

Thr Gln His Gly Thr Val Leu Val Gln Val Lys Tyr Glu Gly Thr Asp

500 505 510

gca cca tgc aag atc ccc ttc tcg tcc caa gat gag aag gga gta acc 2486

Ala Pro Cys Lys Ile Pro Phe Ser Ser Gln Asp Glu Lys Gly Val Thr

515 520 525

cag aat ggg aga ttg ata aca gcc aac ccc ata gtc act gac aaa gaa 2534

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

530 535 540

aaa cca gtc aac att gaa gcg gag cca cct ttt ggt gag agc tac att 2582

Lys Pro Val Asn Ile Glu Ala Glu Pro Pro Phe Gly Glu Ser Tyr Ile

545 550 555

gtg gta gga gca ggt gaa aaa gct ttg aaa cta agc tgg ttc aag aag 2630

Val Val Gly Ala Gly Glu Lys Ala Leu Lys Leu Ser Trp Phe Lys Lys

560 565 570 575

gga agc agt ata ggg aaa atg ttt gaa gca act gcc cgt gga gca cga 2678

Gly Ser Ser Ile Gly Lys Met Phe Glu Ala Thr Ala Arg Gly Ala Arg

580 585 590

agg atg gcc atc ctg gga gac act gca tgg gac ttc ggt tct ata gga 2726

Arg Met Ala Ile Leu Gly Asp Thr Ala Trp Asp Phe Gly Ser Ile Gly

595 600 605

ggg gtg ttc acg tct gtg gga aaa ctg ata cac cag att ttt ggg act 2774

Gly Val Phe Thr Ser Val Gly Lys Leu Ile His Gln Ile Phe Gly Thr

610 615 620

gcg tat gga gtt ttg ttc agc ggt gtt tct tgg acc atg aag ata gga 2822

Ala Tyr Gly Val Leu Phe Ser Gly Val Ser Trp Thr Met Lys Ile Gly

625 630 635

ata ggg att ctg ctg aca tgg cta gga tta aac tca agg agc acg tcc 2870

Ile Gly Ile Leu Leu Thr Trp Leu Gly Leu Asn Ser Arg Ser Thr Ser

640 645 650 655

ctt tca atg acg tgt atc gca gtt ggc atg gtc acg ctg tac cta gga 2918

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

660 665 670

gtc atg gtt cag gcg taataaagat ctgctgtgcc ttctagttgc cagccatctg 2973

Val Met Val Gln Ala

675

ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt 3033

cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg 3093

gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg 3153

atgcggtggg ctctatgggt acccaggtgc tgaagaattg acccggttcc tcctgggcca 3213

gaaagaagca ggcacatccc cttctctgtg acacaccctg tccacgcccc tggttcttag 3273

ttccagcccc actcatagga cactcatagc tcaggagggc tccgccttca atcccacccg 3333

ctaaagtact tggagcggtc tctccctccc tcatcagccc accaaaccaa acctagcctc 3393

caagagtggg aagaaattaa agcaagatag gctattaagt gcagagggag agaaaatgcc 3453

tccaacatgt gaggaagtaa tgagagaaat catagaattt taaggccatg atttaaggcc 3513

atcatggcct aagcttgaaa ggagatagga tcaaagcttg gcgtaatcat ggtcatagct 3573

gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 3633

aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 3693

actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 3753

cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 3813

gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 3873

atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 3933

caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 3993

gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 4053

ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 4113

cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 4173

taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 4233

cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 4293

acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 4353

aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 4413

atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 4473

atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 4533

gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 4593

gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 4653

ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 4713

ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 4773

tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 4833

accatctggc cccagtgctg caatgatacc gcgagaacca cgctcaccgg ctccagattt 4893

atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 4953

cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 5013

tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 5073

tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 5133

gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 5193

agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 5253

aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 5313

gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 5373

tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 5433

gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 5493

tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 5553

aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 5613

catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 5673

acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat 5733

tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gggtcgcgcg 5793

tttcggtgat gacggtgaaa acctctgaca catgcagctc ccgttgacgg tcacagcttg 5853

tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg 5913

gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccataa 5973

aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt 6033

ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agcccgagat 6093

agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa 6153

cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccaa 6213

atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc 6273

ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc 6333

gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac 6393

acccgccgcg cttaatgcgc cgctacaggg cgcgtactat ggttgctttg acgtatgcgg 6453

tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc 6513

aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg 6573

gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca 6633

cgacgttgta aaacgacggc cagtgaattc catggtctca actttc 6679

<210> 8

<211> 676

<212> PRT

<213> 登革病毒

<400> 8

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

1 5 10 15

His Leu Thr Thr Arg Asn Gly Glu Pro His Met Ile Val Ser Arg Gln

20 25 30

Glu Lys Gly Lys Ser Leu Leu Phe Lys Thr Lys Asp Gly Thr Asn Met

35 40 45

Cys Thr Leu Met Ala Met Asp Leu Gly Glu Leu Cys Glu Asp Thr Ile

50 55 60

Thr Tyr Lys Cys Pro Phe Leu Lys Gln Asn Glu Pro Glu Asp Ile Asp

65 70 75 80

Cys Trp Cys Asn Ser Thr Ser Thr Trp Val Thr Tyr Gly Thr Cys Thr

85 90 95

Thr Thr Gly Glu His Arg Arg Glu Lys Arg Ser Val Ala Leu Val Pro

100 105 110

His Val Gly Met Gly Leu Glu Thr Arg Thr Glu Thr Trp Met Ser Ser

115 120 125

Glu Gly Ala Trp Lys His Ala Gln Arg Ile Glu Thr Trp Ile Leu Arg

130 135 140

His Pro Gly Phe Thr Ile Met Ala Ala Ile Leu Ala Tyr Thr Ile Gly

145 150 155 160

Thr Thr His Phe Gln Arg Val Leu Ile Phe Ile Leu Leu Thr Ala Ile

165 170 175

Ala Pro Ser Met Thr Met Arg Cys Ile Gly Ile Ser Asn Arg Asp Phe

180 185 190

Val Glu Gly Val Ser Gly Gly Ser Trp Val Asp Ile Val Leu Glu His

195 200 205

Gly Ser Cys Val Thr Thr Met Ala Lys Asn Lys Pro Thr Leu Asp Phe

210 215 220

Glu Leu Ile Lys Thr Glu Ala Lys Gln Pro Ala Thr Leu Arg Lys Tyr

225 230 235 240

Cys Ile Glu Ala Lys Leu Thr Asn Thr Thr Thr Asp Ser Arg Cys Pro

245 250 255

Thr Gln Gly Glu Pro Thr Leu Asn Glu Glu Gln Asp Lys Arg Phe Val

260 265 270

Cys Lys His Ser Met Val Asp Arg Gly Trp Gly Asn Gly Cys Gly Leu

275 280 285

Phe Gly Lys Gly Gly Ile Val Thr Cys Ala Met Phe Thr Cys Lys Lys

290 295 300

Asn Met Glu Gly Lys Ile Val Gln Pro Glu Asn Leu Glu Tyr Thr Val

305 310 315 320

Val Ile Thr Pro His Ser Gly Glu Glu His Ala Val Gly Asn Asp Thr

325 330 335

Gly Lys His Gly Lys Glu Val Lys Ile Thr Pro Gln Ser Ser Ile Thr

340 345 350

Glu Ala Glu Leu Thr Gly Tyr Gly Thr Val Thr Met Glu Cys Ser Pro

355 360 365

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

370 375 380

Lys Ala Trp Leu Val His Arg Gln Trp Phe Leu Asp Leu Pro Leu Pro

385 390 395 400

Trp Leu Pro Gly Ala Asp Thr Gln Gly Ser Asn Trp Ile Gln Lys Glu

405 410 415

Thr Leu Val Thr Phe Lys Asn Pro His Ala Lys Lys Gln Asp Val Val

420 425 430

Val Leu Gly Ser Gln Glu Gly Ala Met His Thr Ala Leu Thr Gly Ala

435 440 445

Thr Glu Ile Gln Met Ser Ser Gly Asn Leu Leu Phe Thr Gly His Leu

450 455 460

Lys Cys Arg Leu Arg Met Asp Lys Leu Gln Leu Lys Gly Met Ser Tyr

465 470 475 480

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

485 490 495

Gln His Gly Thr Val Leu Val Gln Val Lys Tyr Glu Gly Thr Asp Ala

500 505 510

Pro Cys Lys Ile Pro Phe Ser Ser Gln Asp Glu Lys Gly Val Thr Gln

515 520 525

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

530 535 540

Pro Val Asn Ile Glu Ala Glu Pro Pro Phe Gly Glu Ser Tyr Ile Val

545 550 555 560

Val Gly Ala Gly Glu Lys Ala Leu Lys Leu Ser Trp Phe Lys Lys Gly

565 570 575

Ser Ser Ile Gly Lys Met Phe Glu Ala Thr Ala Arg Gly Ala Arg Arg

580 585 590

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

595 600 605

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

610 615 620

Tyr Gly Val Leu Phe Ser Gly Val Ser Trp Thr Met Lys Ile Gly Ile

625 630 635 640

Gly Ile Leu Leu Thr Trp Leu Gly Leu Asn Ser Arg Ser Thr Ser Leu

645 650 655

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

660 665 670

Met Val Gln Ala

675

<210> 9

<211> 6679

<212> DNA

<213> 登革病毒

<220>

<221> misc_feature

<223> 嵌合体DENV2 prM E (Env F108A)_嵌合DENV1 E

<220>

<221> CDS

<222> (906)..(2933)

<220>

<221> misc_feature

<222> (906)..(2336)

<223> DENV2 prM E区域

<220>

<221> misc_feature

<222> (2337)..(2933)

<223> DENV1区域 (F108A)

<400> 9

gaattcccat tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt 60

ccaacattac cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg 120

gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc 180

ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc 240

atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact 300

gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat 360

gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact 420

tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac 480

atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac 540

gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac 600

tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga 660

gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat 720

agaagacacc gggaccgatc cagcctccgt taacggtgga gggcagtgta gtctgagcag 780

tactcgttgc tgccgcgcgc gccaccagac ataatagctg acagactaac agactgttcc 840

tttccatggg tcttttctgc agtcaccgtc gtcgacacgt gtgatcagat atcgcggccg 900

ccacc atg act gca ggc atg atc atc atg ctg att cca aca gtg atg gcg 950

Met Thr Ala Gly Met Ile Ile Met Leu Ile Pro Thr Val Met Ala

1 5 10 15

ttt cat ctg acc aca cgc aac gga gaa cca cac atg atc gtc agt aga 998

Phe His Leu Thr Thr Arg Asn Gly Glu Pro His Met Ile Val Ser Arg

20 25 30

caa gaa aaa ggg aaa agc ctt ctg ttt aag aca aag gac ggc acg aac 1046

Gln Glu Lys Gly Lys Ser Leu Leu Phe Lys Thr Lys Asp Gly Thr Asn

35 40 45

atg tgt acc ctc atg gcc atg gac ctt ggt gag ttg tgt gaa gac aca 1094

Met Cys Thr Leu Met Ala Met Asp Leu Gly Glu Leu Cys Glu Asp Thr

50 55 60

atc acg tat aaa tgt ccc ttt ctc aag cag aac gaa cca gaa gac ata 1142

Ile Thr Tyr Lys Cys Pro Phe Leu Lys Gln Asn Glu Pro Glu Asp Ile

65 70 75

gat tgt tgg tgc aac tcc acg tcc aca tgg gta act tat ggg aca tgt 1190

Asp Cys Trp Cys Asn Ser Thr Ser Thr Trp Val Thr Tyr Gly Thr Cys

80 85 90 95

acc acc aca gga gag cac aga aga gaa aaa aga tca gtg gcg ctt gtt 1238

Thr Thr Thr Gly Glu His Arg Arg Glu Lys Arg Ser Val Ala Leu Val

100 105 110

cca cac gtg gga atg gga ttg gag aca cga act gaa aca tgg atg tca 1286

Pro His Val Gly Met Gly Leu Glu Thr Arg Thr Glu Thr Trp Met Ser

115 120 125

tca gaa ggg gcc tgg aaa cat gcc cag aga att gaa act tgg att ctg 1334

Ser Glu Gly Ala Trp Lys His Ala Gln Arg Ile Glu Thr Trp Ile Leu

130 135 140

aga cat cca ggc ttt acc ata atg gcc gca atc ctg gca tac acc ata 1382

Arg His Pro Gly Phe Thr Ile Met Ala Ala Ile Leu Ala Tyr Thr Ile

145 150 155

gga acg acg cat ttc caa aga gtc ctg ata ttc atc cta ctg aca gcc 1430

Gly Thr Thr His Phe Gln Arg Val Leu Ile Phe Ile Leu Leu Thr Ala

160 165 170 175

atc gct cct tca atg aca atg cgc tgc ata gga ata tca aat agg gac 1478

Ile Ala Pro Ser Met Thr Met Arg Cys Ile Gly Ile Ser Asn Arg Asp

180 185 190

ttt gtg gaa gga gtg tca gga ggg agt tgg gtt gac ata gtt tta gaa 1526

Phe Val Glu Gly Val Ser Gly Gly Ser Trp Val Asp Ile Val Leu Glu

195 200 205

cat gga agt tgt gtg acg acg atg gca aaa aat aaa cca aca ctg gac 1574

His Gly Ser Cys Val Thr Thr Met Ala Lys Asn Lys Pro Thr Leu Asp

210 215 220

ttt gaa ctg ata aaa aca gaa gcc aaa caa ccc gcc acc tta agg aag 1622

Phe Glu Leu Ile Lys Thr Glu Ala Lys Gln Pro Ala Thr Leu Arg Lys

225 230 235

tac tgt ata gag gct aaa ctg acc aac acg aca aca gac tcg cgc tgc 1670

Tyr Cys Ile Glu Ala Lys Leu Thr Asn Thr Thr Thr Asp Ser Arg Cys

240 245 250 255

cca aca caa ggg gaa ccc acc ctg aat gaa gag cag gac aaa agg ttt 1718

Pro Thr Gln Gly Glu Pro Thr Leu Asn Glu Glu Gln Asp Lys Arg Phe

260 265 270

gtc tgc aaa cat tcc atg gta gac aga gga tgg gga aat gga tgt gga 1766

Val Cys Lys His Ser Met Val Asp Arg Gly Trp Gly Asn Gly Cys Gly

275 280 285

tta gct gga aaa gga ggc atc gtg acc tgt gcc atg ttc aca tgc aaa 1814

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

290 295 300

aag aac atg gag gga aaa att gtg cag cca gaa aac ctg gaa tac act 1862

Lys Asn Met Glu Gly Lys Ile Val Gln Pro Glu Asn Leu Glu Tyr Thr

305 310 315

gtc gtt ata aca cct cat tca ggg gaa gaa cat gca gtc gga aat gac 1910

Val Val Ile Thr Pro His Ser Gly Glu Glu His Ala Val Gly Asn Asp

320 325 330 335

aca gga aaa cat ggt aaa gaa gtc aag ata aca cca cag agc tcc atc 1958

Thr Gly Lys His Gly Lys Glu Val Lys Ile Thr Pro Gln Ser Ser Ile

340 345 350

aca gag gcg gaa ctg aca ggc tat ggc act gtt acg atg gag tgc tct 2006

Thr Glu Ala Glu Leu Thr Gly Tyr Gly Thr Val Thr Met Glu Cys Ser

355 360 365

cca aga acg ggc ctc gac ttc aat gag atg gtg ttg ctg caa atg aaa 2054

Pro Arg Thr Gly Leu Asp Phe Asn Glu Met Val Leu Leu Gln Met Lys

370 375 380

gac aaa gct tgg ctg gtg cac aga caa tgg ttc cta gac cta ccg ttg 2102

Asp Lys Ala Trp Leu Val His Arg Gln Trp Phe Leu Asp Leu Pro Leu

385 390 395

cca tgg ctg ccc gga gca gac aca caa gga tca aat tgg ata cag aaa 2150

Pro Trp Leu Pro Gly Ala Asp Thr Gln Gly Ser Asn Trp Ile Gln Lys

400 405 410 415

gag aca ctg gtc acc ttc aaa aat ccc cat gcg aaa aaa cag gat gtt 2198

Glu Thr Leu Val Thr Phe Lys Asn Pro His Ala Lys Lys Gln Asp Val

420 425 430

gtt gtc tta ggc tcc caa gag ggg gcc atg cat aca gca ctc aca ggg 2246

Val Val Leu Gly Ser Gln Glu Gly Ala Met His Thr Ala Leu Thr Gly

435 440 445

gct acg gaa atc cag atg tca tca gga aac ctg ctg ttc aca gga cat 2294

Ala Thr Glu Ile Gln Met Ser Ser Gly Asn Leu Leu Phe Thr Gly His

450 455 460

ctt aag tgc agg ctg aga atg gac aaa tta caa ctt aaa ggg atg tca 2342

Leu Lys Cys Arg Leu Arg Met Asp Lys Leu Gln Leu Lys Gly Met Ser

465 470 475

tat gta atg tgc aca ggg tca ttc aag tta gag aag gaa gtg gct gag 2390

Tyr Val Met Cys Thr Gly Ser Phe Lys Leu Glu Lys Glu Val Ala Glu

480 485 490 495

acc cag cat gga act gtt cta gtg cag gtt aaa tac gaa gga aca gat 2438

Thr Gln His Gly Thr Val Leu Val Gln Val Lys Tyr Glu Gly Thr Asp

500 505 510

gca cca tgc aag atc ccc ttc tcg tcc caa gat gag aag gga gta acc 2486

Ala Pro Cys Lys Ile Pro Phe Ser Ser Gln Asp Glu Lys Gly Val Thr

515 520 525

cag aat ggg aga ttg ata aca gcc aac ccc ata gtc act gac aaa gaa 2534

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

530 535 540

aaa cca gtc aac att gaa gcg gag cca cct ttt ggt gag agc tac att 2582

Lys Pro Val Asn Ile Glu Ala Glu Pro Pro Phe Gly Glu Ser Tyr Ile

545 550 555

gtg gta gga gca ggt gaa aaa gct ttg aaa cta agc tgg ttc aag aag 2630

Val Val Gly Ala Gly Glu Lys Ala Leu Lys Leu Ser Trp Phe Lys Lys

560 565 570 575

gga agc agt ata ggg aaa atg ttt gaa gca act gcc cgt gga gca cga 2678

Gly Ser Ser Ile Gly Lys Met Phe Glu Ala Thr Ala Arg Gly Ala Arg

580 585 590

agg atg gcc atc ctg gga gac act gca tgg gac ttc ggt tct ata gga 2726

Arg Met Ala Ile Leu Gly Asp Thr Ala Trp Asp Phe Gly Ser Ile Gly

595 600 605

ggg gtg ttc acg tct gtg gga aaa ctg ata cac cag att ttt ggg act 2774

Gly Val Phe Thr Ser Val Gly Lys Leu Ile His Gln Ile Phe Gly Thr

610 615 620

gcg tat gga gtt ttg ttc agc ggt gtt tct tgg acc atg aag ata gga 2822

Ala Tyr Gly Val Leu Phe Ser Gly Val Ser Trp Thr Met Lys Ile Gly

625 630 635

ata ggg att ctg ctg aca tgg cta gga tta aac tca agg agc acg tcc 2870

Ile Gly Ile Leu Leu Thr Trp Leu Gly Leu Asn Ser Arg Ser Thr Ser

640 645 650 655

ctt tca atg acg tgt atc gca gtt ggc atg gtc acg ctg tac cta gga 2918

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

660 665 670

gtc atg gtt cag gcg taataaagat ctgctgtgcc ttctagttgc cagccatctg 2973

Val Met Val Gln Ala

675

ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt 3033

cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg 3093

gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg 3153

atgcggtggg ctctatgggt acccaggtgc tgaagaattg acccggttcc tcctgggcca 3213

gaaagaagca ggcacatccc cttctctgtg acacaccctg tccacgcccc tggttcttag 3273

ttccagcccc actcatagga cactcatagc tcaggagggc tccgccttca atcccacccg 3333

ctaaagtact tggagcggtc tctccctccc tcatcagccc accaaaccaa acctagcctc 3393

caagagtggg aagaaattaa agcaagatag gctattaagt gcagagggag agaaaatgcc 3453

tccaacatgt gaggaagtaa tgagagaaat catagaattt taaggccatg atttaaggcc 3513

atcatggcct aagcttgaaa ggagatagga tcaaagcttg gcgtaatcat ggtcatagct 3573

gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 3633

aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 3693

actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 3753

cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 3813

gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 3873

atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 3933

caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 3993

gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 4053

ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 4113

cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 4173

taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 4233

cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 4293

acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 4353

aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 4413

atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 4473

atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 4533

gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 4593

gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 4653

ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 4713

ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 4773

tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 4833

accatctggc cccagtgctg caatgatacc gcgagaacca cgctcaccgg ctccagattt 4893

atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 4953

cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 5013

tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 5073

tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 5133

gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 5193

agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 5253

aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 5313

gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 5373

tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 5433

gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 5493

tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 5553

aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 5613

catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 5673

acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat 5733

tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gggtcgcgcg 5793

tttcggtgat gacggtgaaa acctctgaca catgcagctc ccgttgacgg tcacagcttg 5853

tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg 5913

gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccataa 5973

aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt 6033

ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agcccgagat 6093

agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa 6153

cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccaa 6213

atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc 6273

ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc 6333

gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac 6393

acccgccgcg cttaatgcgc cgctacaggg cgcgtactat ggttgctttg acgtatgcgg 6453

tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc 6513

aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg 6573

gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca 6633

cgacgttgta aaacgacggc cagtgaattc catggtctca actttc 6679

<210> 10

<211> 676

<212> PRT

<213> 登革病毒

<400> 10

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

1 5 10 15

His Leu Thr Thr Arg Asn Gly Glu Pro His Met Ile Val Ser Arg Gln

20 25 30

Glu Lys Gly Lys Ser Leu Leu Phe Lys Thr Lys Asp Gly Thr Asn Met

35 40 45

Cys Thr Leu Met Ala Met Asp Leu Gly Glu Leu Cys Glu Asp Thr Ile

50 55 60

Thr Tyr Lys Cys Pro Phe Leu Lys Gln Asn Glu Pro Glu Asp Ile Asp

65 70 75 80

Cys Trp Cys Asn Ser Thr Ser Thr Trp Val Thr Tyr Gly Thr Cys Thr

85 90 95

Thr Thr Gly Glu His Arg Arg Glu Lys Arg Ser Val Ala Leu Val Pro

100 105 110

His Val Gly Met Gly Leu Glu Thr Arg Thr Glu Thr Trp Met Ser Ser

115 120 125

Glu Gly Ala Trp Lys His Ala Gln Arg Ile Glu Thr Trp Ile Leu Arg

130 135 140

His Pro Gly Phe Thr Ile Met Ala Ala Ile Leu Ala Tyr Thr Ile Gly

145 150 155 160

Thr Thr His Phe Gln Arg Val Leu Ile Phe Ile Leu Leu Thr Ala Ile

165 170 175

Ala Pro Ser Met Thr Met Arg Cys Ile Gly Ile Ser Asn Arg Asp Phe

180 185 190

Val Glu Gly Val Ser Gly Gly Ser Trp Val Asp Ile Val Leu Glu His

195 200 205

Gly Ser Cys Val Thr Thr Met Ala Lys Asn Lys Pro Thr Leu Asp Phe

210 215 220

Glu Leu Ile Lys Thr Glu Ala Lys Gln Pro Ala Thr Leu Arg Lys Tyr

225 230 235 240

Cys Ile Glu Ala Lys Leu Thr Asn Thr Thr Thr Asp Ser Arg Cys Pro

245 250 255

Thr Gln Gly Glu Pro Thr Leu Asn Glu Glu Gln Asp Lys Arg Phe Val

260 265 270

Cys Lys His Ser Met Val Asp Arg Gly Trp Gly Asn Gly Cys Gly Leu

275 280 285

Ala Gly Lys Gly Gly Ile Val Thr Cys Ala Met Phe Thr Cys Lys Lys

290 295 300

Asn Met Glu Gly Lys Ile Val Gln Pro Glu Asn Leu Glu Tyr Thr Val

305 310 315 320

Val Ile Thr Pro His Ser Gly Glu Glu His Ala Val Gly Asn Asp Thr

325 330 335

Gly Lys His Gly Lys Glu Val Lys Ile Thr Pro Gln Ser Ser Ile Thr

340 345 350

Glu Ala Glu Leu Thr Gly Tyr Gly Thr Val Thr Met Glu Cys Ser Pro

355 360 365

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

370 375 380

Lys Ala Trp Leu Val His Arg Gln Trp Phe Leu Asp Leu Pro Leu Pro

385 390 395 400

Trp Leu Pro Gly Ala Asp Thr Gln Gly Ser Asn Trp Ile Gln Lys Glu

405 410 415

Thr Leu Val Thr Phe Lys Asn Pro His Ala Lys Lys Gln Asp Val Val

420 425 430

Val Leu Gly Ser Gln Glu Gly Ala Met His Thr Ala Leu Thr Gly Ala

435 440 445

Thr Glu Ile Gln Met Ser Ser Gly Asn Leu Leu Phe Thr Gly His Leu

450 455 460

Lys Cys Arg Leu Arg Met Asp Lys Leu Gln Leu Lys Gly Met Ser Tyr

465 470 475 480

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

485 490 495

Gln His Gly Thr Val Leu Val Gln Val Lys Tyr Glu Gly Thr Asp Ala

500 505 510

Pro Cys Lys Ile Pro Phe Ser Ser Gln Asp Glu Lys Gly Val Thr Gln

515 520 525

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

530 535 540

Pro Val Asn Ile Glu Ala Glu Pro Pro Phe Gly Glu Ser Tyr Ile Val

545 550 555 560

Val Gly Ala Gly Glu Lys Ala Leu Lys Leu Ser Trp Phe Lys Lys Gly

565 570 575

Ser Ser Ile Gly Lys Met Phe Glu Ala Thr Ala Arg Gly Ala Arg Arg

580 585 590

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

595 600 605

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

610 615 620

Tyr Gly Val Leu Phe Ser Gly Val Ser Trp Thr Met Lys Ile Gly Ile

625 630 635 640

Gly Ile Leu Leu Thr Trp Leu Gly Leu Asn Ser Arg Ser Thr Ser Leu

645 650 655

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

660 665 670

Met Val Gln Ala

675

<210> 11

<211> 3770

<212> DNA

<213> 登革病毒

<220>

<221> misc_feature

<223> DENV2衣壳蛋白R85A K86A突变体

<220>

<221> CDS

<222> (1046)..(1345)

<400> 11

tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 60

cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 120

ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 180

accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 240

attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 300

tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 360

tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gacgcgtttg acattgatta 420

ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 480

ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 540

ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 600

cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 660

atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 720

cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 780

attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 840

cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 900

caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 960

cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gataactgca 1020

ggtcgacgat atcgcggccg ccacc atg aat gac caa cgg aaa aag gcg aga 1072

Met Asn Asp Gln Arg Lys Lys Ala Arg

1 5

aac acg cct ttc aat atg ctg aaa cgc gag aga aac cgc gtg tca act 1120

Asn Thr Pro Phe Asn Met Leu Lys Arg Glu Arg Asn Arg Val Ser Thr

10 15 20 25

gta caa cag ttg aca aag aga ttc tca ctt gga atg ctg cag gga cga 1168

Val Gln Gln Leu Thr Lys Arg Phe Ser Leu Gly Met Leu Gln Gly Arg

30 35 40

gga cca cta aaa ttg ttc atg gcc ctg gtg gca ttc ctt cgt ttc cta 1216

Gly Pro Leu Lys Leu Phe Met Ala Leu Val Ala Phe Leu Arg Phe Leu

45 50 55

aca atc cca cca aca gca ggg ata tta aaa aga tgg gga aca att aaa 1264

Thr Ile Pro Pro Thr Ala Gly Ile Leu Lys Arg Trp Gly Thr Ile Lys

60 65 70

aaa tca aag gct att aat gtt ctg aga ggc ttc gcg gca gag att gga 1312

Lys Ser Lys Ala Ile Asn Val Leu Arg Gly Phe Ala Ala Glu Ile Gly

75 80 85

agg atg ctg aat atc tta aac agg aga cgt aga taataaagat ctgctgtgcc 1365

Arg Met Leu Asn Ile Leu Asn Arg Arg Arg Arg

90 95 100

ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 1425

tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 1485

gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 1545

caatagcagg catgctgggg atgcggtggg ctctatggct cgagcatggt catagctgtt 1605

tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa 1665

gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact 1725

gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 1785

ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg 1845

ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 1905

cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag 1965

gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 2025

tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 2085

ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 2145

atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 2205

gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 2265

tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 2325

cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 2385

cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt 2445

tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 2505

cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 2565

cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 2625

gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 2685

gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg 2745

gtctgacagt tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg 2805

attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag 2865

gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc 2925

aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg 2985

agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc 3045

aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat 3105

tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac 3165

aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga 3225

atcaggatat tcttctaata cctggaatgc tgttttccca gggatcgcag tggtgagtaa 3285

ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt 3345

cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg 3405

tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga 3465

ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt 3525

taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcatactct tcctttttca 3585

atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 3645

ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 3705

ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt 3765

tcgtc 3770

<210> 12

<211> 100

<212> PRT

<213> 登革病毒

<400> 12

Met Asn Asp Gln Arg Lys Lys Ala Arg Asn Thr Pro Phe Asn Met Leu

1 5 10 15

Lys Arg Glu Arg Asn Arg Val Ser Thr Val Gln Gln Leu Thr Lys Arg

20 25 30

Phe Ser Leu Gly Met Leu Gln Gly Arg Gly Pro Leu Lys Leu Phe Met

35 40 45

Ala Leu Val Ala Phe Leu Arg Phe Leu Thr Ile Pro Pro Thr Ala Gly

50 55 60

Ile Leu Lys Arg Trp Gly Thr Ile Lys Lys Ser Lys Ala Ile Asn Val

65 70 75 80

Leu Arg Gly Phe Ala Ala Glu Ile Gly Arg Met Leu Asn Ile Leu Asn

85 90 95

Arg Arg Arg Arg

100

<210> 13

<211> 6910

<212> DNA

<213> 登革病毒

<220>

<221> misc_feature

<223> DENV1 prM E (F108A K246M)

<220>

<221> CDS

<222> (1137)..(3164)

<400> 13

gaattcccat tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt 60

ccaacattac cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg 120

gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc 180

ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc 240

atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact 300

gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat 360

gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact 420

tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac 480

atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac 540

gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac 600

tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga 660

gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat 720

agaagacacc gggaccgatc cagcctccat cggctcgcat ctctccttca cgcgcccgcc 780

gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg 840

gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc 900

tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga 960

ccctgcttgc tcaactctag ttaacggtgg agggcagtgt agtctgagca gtactcgttg 1020

ctgccgcgcg cgccaccaga cataatagct gacagactaa cagactgttc ctttccatgg 1080

gtcttttctg cagtcaccgt cgtcgacacg tgtgatcaga tatcgcggcc gccacc atg 1139

Met

1

tcg gtg acc atg ctc ctc atg ctg ctg ccc aca gcc ctg gcg ttc cat 1187

Ser Val Thr Met Leu Leu Met Leu Leu Pro Thr Ala Leu Ala Phe His

5 10 15

ctg acc acc cga ggg gga gag ccg cac atg ata gtt agc aag cag gaa 1235

Leu Thr Thr Arg Gly Gly Glu Pro His Met Ile Val Ser Lys Gln Glu

20 25 30

aga gga aaa tca ctt ttg ttt aag acc tct gca ggt gtc aac atg tgc 1283

Arg Gly Lys Ser Leu Leu Phe Lys Thr Ser Ala Gly Val Asn Met Cys

35 40 45

acc ctt att gca atg gat ttg gga gag tta tgt gag gac aca atg acc 1331

Thr Leu Ile Ala Met Asp Leu Gly Glu Leu Cys Glu Asp Thr Met Thr

50 55 60 65

tac aaa tgc ccc cgg atc act gag acg gaa cca gat gac gtt gac tgt 1379

Tyr Lys Cys Pro Arg Ile Thr Glu Thr Glu Pro Asp Asp Val Asp Cys

70 75 80

tgg tgc aat gcc acg gag aca tgg gtg acc tat gga aca tgt tct caa 1427

Trp Cys Asn Ala Thr Glu Thr Trp Val Thr Tyr Gly Thr Cys Ser Gln

85 90 95

act ggt gaa cac cga cga gac aaa cgt tcc gtc gca ctg gca cca cac 1475

Thr Gly Glu His Arg Arg Asp Lys Arg Ser Val Ala Leu Ala Pro His

100 105 110

gta ggg ctt ggt cta gaa aca aga acc gaa acg tgg atg tcc tct gaa 1523

Val Gly Leu Gly Leu Glu Thr Arg Thr Glu Thr Trp Met Ser Ser Glu

115 120 125

ggc gct tgg aaa caa ata caa aaa gtg gag acc tgg gct ctg aga cac 1571

Gly Ala Trp Lys Gln Ile Gln Lys Val Glu Thr Trp Ala Leu Arg His

130 135 140 145

cca gga ttc acg gtg ata gcc ctt ttt cta gca cat gcc ata gga aca 1619

Pro Gly Phe Thr Val Ile Ala Leu Phe Leu Ala His Ala Ile Gly Thr

150 155 160

tcc atc acc cag aaa ggg atc att ttt att ttg ctg atg ctg gta act 1667

Ser Ile Thr Gln Lys Gly Ile Ile Phe Ile Leu Leu Met Leu Val Thr

165 170 175

cca tcc atg gcc atg cgg tgc gtg gga ata ggc aac aga gac ttc gtg 1715

Pro Ser Met Ala Met Arg Cys Val Gly Ile Gly Asn Arg Asp Phe Val

180 185 190

gaa gga ctg tca gga gct acg tgg gtg gat gtg gta ctg gag cat gga 1763

Glu Gly Leu Ser Gly Ala Thr Trp Val Asp Val Val Leu Glu His Gly

195 200 205

agt tgc gtc act acc atg gca aaa gac aaa cca aca ctg gac att gaa 1811

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

210 215 220 225

ctc ttg aag acg gag gtc aca aac cct gcc gtc ctg cgc aaa ctg tgc 1859

Leu Leu Lys Thr Glu Val Thr Asn Pro Ala Val Leu Arg Lys Leu Cys

230 235 240

att gaa gct aaa ata tca aac acc acc acc gat tcg aga tgt cca aca 1907

Ile Glu Ala Lys Ile Ser Asn Thr Thr Thr Asp Ser Arg Cys Pro Thr

245 250 255

caa gga gaa gcc acg ctg gtg gaa gaa cag gac acg aac ttt gtg tgt 1955

Gln Gly Glu Ala Thr Leu Val Glu Glu Gln Asp Thr Asn Phe Val Cys

260 265 270

cga cga acg ttc gtg gac aga ggc tgg ggc aat ggt tgt ggg cta gcc 2003

Arg Arg Thr Phe Val Asp Arg Gly Trp Gly Asn Gly Cys Gly Leu Ala

275 280 285

gga aaa ggt agc tta ata acg tgt gct aag ttt aag tgt gtg aca aaa 2051

Gly Lys Gly Ser Leu Ile Thr Cys Ala Lys Phe Lys Cys Val Thr Lys

290 295 300 305

ctg gaa gga aag ata gtc caa tat gaa aac tta aaa tat tca gtg ata 2099

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

310 315 320

gtc acc gta cac act gga gac cag cac caa gtt gga aat gag acc aca 2147

Val Thr Val His Thr Gly Asp Gln His Gln Val Gly Asn Glu Thr Thr

325 330 335

gaa cat gga aca act gca acc ata aca cct caa gct ccc acg tcg gaa 2195

Glu His Gly Thr Thr Ala Thr Ile Thr Pro Gln Ala Pro Thr Ser Glu

340 345 350

ata cag ctg aca gac tac gga gct cta aca ttg gat tgt tca cct aga 2243

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

355 360 365

aca ggg cta gac ttt aat gag atg gtg ttg ttg aca atg gaa aat aaa 2291

Thr Gly Leu Asp Phe Asn Glu Met Val Leu Leu Thr Met Glu Asn Lys

370 375 380 385

tca tgg ctc gtc cac aaa caa tgg ttt cta gac tta cca ctg cct tgg 2339

Ser Trp Leu Val His Lys Gln Trp Phe Leu Asp Leu Pro Leu Pro Trp

390 395 400

acc tcg ggg gct tca aca tcc caa gag act tgg aat aga caa gac ttg 2387

Thr Ser Gly Ala Ser Thr Ser Gln Glu Thr Trp Asn Arg Gln Asp Leu

405 410 415

ctg gtc aca ttt aag aca gct cat gca aaa aag cag gaa gta gtc gta 2435

Leu Val Thr Phe Lys Thr Ala His Ala Lys Lys Gln Glu Val Val Val

420 425 430

cta gga tca caa gaa gga gca atg cac act gcg ttg act gga gcg aca 2483

Leu Gly Ser Gln Glu Gly Ala Met His Thr Ala Leu Thr Gly Ala Thr

435 440 445

gaa atc caa acg tct gga acg aca aca att ttt gca gga cac ctg aaa 2531

Glu Ile Gln Thr Ser Gly Thr Thr Thr Ile Phe Ala Gly His Leu Lys

450 455 460 465

tgc aga cta aaa atg gat aaa ctg act tta aaa ggg atg tca tat gta 2579

Cys Arg Leu Lys Met Asp Lys Leu Thr Leu Lys Gly Met Ser Tyr Val

470 475 480

atg tgc aca ggg tca ttc aag tta gag aag gaa gtg gct gag acc cag 2627

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

485 490 495

cat gga act gtt cta gtg cag gtt aaa tac gaa gga aca gat gca cca 2675

His Gly Thr Val Leu Val Gln Val Lys Tyr Glu Gly Thr Asp Ala Pro

500 505 510

tgc aag atc ccc ttc tcg tcc caa gat gag aag gga gta acc cag aat 2723

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

515 520 525

ggg aga ttg ata aca gcc aac ccc ata gtc act gac aaa gaa aaa cca 2771

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

530 535 540 545

gtc aac att gaa gcg gag cca cct ttt ggt gag agc tac att gtg gta 2819

Val Asn Ile Glu Ala Glu Pro Pro Phe Gly Glu Ser Tyr Ile Val Val

550 555 560

gga gca ggt gaa aaa gct ttg aaa cta agc tgg ttc aag aag gga agc 2867

Gly Ala Gly Glu Lys Ala Leu Lys Leu Ser Trp Phe Lys Lys Gly Ser

565 570 575

agt ata ggg aaa atg ttt gaa gca act gcc cgt gga gca cga agg atg 2915

Ser Ile Gly Lys Met Phe Glu Ala Thr Ala Arg Gly Ala Arg Arg Met

580 585 590

gcc atc ctg gga gac act gca tgg gac ttc ggt tct ata gga ggg gtg 2963

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

595 600 605

ttc acg tct gtg gga aaa ctg ata cac cag att ttt ggg act gcg tat 3011

Phe Thr Ser Val Gly Lys Leu Ile His Gln Ile Phe Gly Thr Ala Tyr

610 615 620 625

gga gtt ttg ttc agc ggt gtt tct tgg acc atg aag ata gga ata ggg 3059

Gly Val Leu Phe Ser Gly Val Ser Trp Thr Met Lys Ile Gly Ile Gly

630 635 640

att ctg ctg aca tgg cta gga tta aac tca agg agc acg tcc ctt tca 3107

Ile Leu Leu Thr Trp Leu Gly Leu Asn Ser Arg Ser Thr Ser Leu Ser

645 650 655

atg acg tgt atc gca gtt ggc atg gtc acg ctg tac cta gga gtc atg 3155

Met Thr Cys Ile Ala Val Gly Met Val Thr Leu Tyr Leu Gly Val Met

660 665 670

gtt cag gcg taataaagat ctgctgtgcc ttctagttgc cagccatctg 3204

Val Gln Ala

675

ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt 3264

cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg 3324

gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg 3384

atgcggtggg ctctatgggt acccaggtgc tgaagaattg acccggttcc tcctgggcca 3444

gaaagaagca ggcacatccc cttctctgtg acacaccctg tccacgcccc tggttcttag 3504

ttccagcccc actcatagga cactcatagc tcaggagggc tccgccttca atcccacccg 3564

ctaaagtact tggagcggtc tctccctccc tcatcagccc accaaaccaa acctagcctc 3624

caagagtggg aagaaattaa agcaagatag gctattaagt gcagagggag agaaaatgcc 3684

tccaacatgt gaggaagtaa tgagagaaat catagaattt taaggccatg atttaaggcc 3744

atcatggcct aagcttgaaa ggagatagga tcaaagcttg gcgtaatcat ggtcatagct 3804

gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 3864

aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 3924

actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 3984

cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 4044

gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 4104

atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 4164

caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 4224

gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 4284

ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 4344

cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 4404

taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 4464

cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 4524

acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 4584

aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 4644

atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 4704

atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 4764

gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 4824

gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 4884

ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 4944

ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 5004

tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 5064

accatctggc cccagtgctg caatgatacc gcgagaacca cgctcaccgg ctccagattt 5124

atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 5184

cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 5244

tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 5304

tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 5364

gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 5424

agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 5484

aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 5544

gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 5604

tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 5664

gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 5724

tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 5784

aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 5844

catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 5904

acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat 5964

tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gggtcgcgcg 6024

tttcggtgat gacggtgaaa acctctgaca catgcagctc ccgttgacgg tcacagcttg 6084

tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg 6144

gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccataa 6204

aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt 6264

ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agcccgagat 6324

agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa 6384

cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccaa 6444

atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc 6504

ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc 6564

gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac 6624

acccgccgcg cttaatgcgc cgctacaggg cgcgtactat ggttgctttg acgtatgcgg 6684

tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc 6744

aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg 6804

gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca 6864

cgacgttgta aaacgacggc cagtgaattc catggtctca actttc 6910

<210> 14

<211> 676

<212> PRT

<213> 登革病毒

<400> 14

Met Ser Val Thr Met Leu Leu Met Leu Leu Pro Thr Ala Leu Ala Phe

1 5 10 15

His Leu Thr Thr Arg Gly Gly Glu Pro His Met Ile Val Ser Lys Gln

20 25 30

Glu Arg Gly Lys Ser Leu Leu Phe Lys Thr Ser Ala Gly Val Asn Met

35 40 45

Cys Thr Leu Ile Ala Met Asp Leu Gly Glu Leu Cys Glu Asp Thr Met

50 55 60

Thr Tyr Lys Cys Pro Arg Ile Thr Glu Thr Glu Pro Asp Asp Val Asp

65 70 75 80

Cys Trp Cys Asn Ala Thr Glu Thr Trp Val Thr Tyr Gly Thr Cys Ser

85 90 95

Gln Thr Gly Glu His Arg Arg Asp Lys Arg Ser Val Ala Leu Ala Pro

100 105 110

His Val Gly Leu Gly Leu Glu Thr Arg Thr Glu Thr Trp Met Ser Ser

115 120 125

Glu Gly Ala Trp Lys Gln Ile Gln Lys Val Glu Thr Trp Ala Leu Arg

130 135 140

His Pro Gly Phe Thr Val Ile Ala Leu Phe Leu Ala His Ala Ile Gly

145 150 155 160

Thr Ser Ile Thr Gln Lys Gly Ile Ile Phe Ile Leu Leu Met Leu Val

165 170 175

Thr Pro Ser Met Ala Met Arg Cys Val Gly Ile Gly Asn Arg Asp Phe

180 185 190

Val Glu Gly Leu Ser Gly Ala Thr Trp Val Asp Val Val Leu Glu His

195 200 205

Gly Ser Cys Val Thr Thr Met Ala Lys Asp Lys Pro Thr Leu Asp Ile

210 215 220

Glu Leu Leu Lys Thr Glu Val Thr Asn Pro Ala Val Leu Arg Lys Leu

225 230 235 240

Cys Ile Glu Ala Lys Ile Ser Asn Thr Thr Thr Asp Ser Arg Cys Pro

245 250 255

Thr Gln Gly Glu Ala Thr Leu Val Glu Glu Gln Asp Thr Asn Phe Val

260 265 270

Cys Arg Arg Thr Phe Val Asp Arg Gly Trp Gly Asn Gly Cys Gly Leu

275 280 285

Ala Gly Lys Gly Ser Leu Ile Thr Cys Ala Lys Phe Lys Cys Val Thr

290 295 300

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

305 310 315 320

Ile Val Thr Val His Thr Gly Asp Gln His Gln Val Gly Asn Glu Thr

325 330 335

Thr Glu His Gly Thr Thr Ala Thr Ile Thr Pro Gln Ala Pro Thr Ser

340 345 350

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

355 360 365

Arg Thr Gly Leu Asp Phe Asn Glu Met Val Leu Leu Thr Met Glu Asn

370 375 380

Lys Ser Trp Leu Val His Lys Gln Trp Phe Leu Asp Leu Pro Leu Pro

385 390 395 400

Trp Thr Ser Gly Ala Ser Thr Ser Gln Glu Thr Trp Asn Arg Gln Asp

405 410 415

Leu Leu Val Thr Phe Lys Thr Ala His Ala Lys Lys Gln Glu Val Val

420 425 430

Val Leu Gly Ser Gln Glu Gly Ala Met His Thr Ala Leu Thr Gly Ala

435 440 445

Thr Glu Ile Gln Thr Ser Gly Thr Thr Thr Ile Phe Ala Gly His Leu

450 455 460

Lys Cys Arg Leu Lys Met Asp Lys Leu Thr Leu Lys Gly Met Ser Tyr

465 470 475 480

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

485 490 495

Gln His Gly Thr Val Leu Val Gln Val Lys Tyr Glu Gly Thr Asp Ala

500 505 510

Pro Cys Lys Ile Pro Phe Ser Ser Gln Asp Glu Lys Gly Val Thr Gln

515 520 525

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

530 535 540

Pro Val Asn Ile Glu Ala Glu Pro Pro Phe Gly Glu Ser Tyr Ile Val

545 550 555 560

Val Gly Ala Gly Glu Lys Ala Leu Lys Leu Ser Trp Phe Lys Lys Gly

565 570 575

Ser Ser Ile Gly Lys Met Phe Glu Ala Thr Ala Arg Gly Ala Arg Arg

580 585 590

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

595 600 605

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

610 615 620

Tyr Gly Val Leu Phe Ser Gly Val Ser Trp Thr Met Lys Ile Gly Ile

625 630 635 640

Gly Ile Leu Leu Thr Trp Leu Gly Leu Asn Ser Arg Ser Thr Ser Leu

645 650 655

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

660 665 670

Met Val Gln Ala

675

<210> 15

<211> 6910

<212> DNA

<213> 登革病毒

<220>

<221> misc_feature

<223> DENV1 prM E (F108A K203N) VLP D01

<220>

<221> CDS

<222> (1137)..(3164)

<400> 15

gaattcccat tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt 60

ccaacattac cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg 120

gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc 180

ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc 240

atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact 300

gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat 360

gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact 420

tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac 480

atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac 540

gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac 600

tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga 660

gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat 720

agaagacacc gggaccgatc cagcctccat cggctcgcat ctctccttca cgcgcccgcc 780

gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg 840

gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc 900

tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga 960

ccctgcttgc tcaactctag ttaacggtgg agggcagtgt agtctgagca gtactcgttg 1020

ctgccgcgcg cgccaccaga cataatagct gacagactaa cagactgttc ctttccatgg 1080

gtcttttctg cagtcaccgt cgtcgacacg tgtgatcaga tatcgcggcc gccacc atg 1139

Met

1

tcg gtg acc atg ctc ctc atg ctg ctg ccc aca gcc ctg gcg ttc cat 1187

Ser Val Thr Met Leu Leu Met Leu Leu Pro Thr Ala Leu Ala Phe His

5 10 15

ctg acc acc cga ggg gga gag ccg cac atg ata gtt agc aag cag gaa 1235

Leu Thr Thr Arg Gly Gly Glu Pro His Met Ile Val Ser Lys Gln Glu

20 25 30

aga gga aaa tca ctt ttg ttt aag acc tct gca ggt gtc aac atg tgc 1283

Arg Gly Lys Ser Leu Leu Phe Lys Thr Ser Ala Gly Val Asn Met Cys

35 40 45

acc ctt att gca atg gat ttg gga gag tta tgt gag gac aca atg acc 1331

Thr Leu Ile Ala Met Asp Leu Gly Glu Leu Cys Glu Asp Thr Met Thr

50 55 60 65

tac aaa tgc ccc cgg atc act gag acg gaa cca gat gac gtt gac tgt 1379

Tyr Lys Cys Pro Arg Ile Thr Glu Thr Glu Pro Asp Asp Val Asp Cys

70 75 80

tgg tgc aat gcc acg gag aca tgg gtg acc tat gga aca tgt tct caa 1427

Trp Cys Asn Ala Thr Glu Thr Trp Val Thr Tyr Gly Thr Cys Ser Gln

85 90 95

act ggt gaa cac cga cga gac aaa cgt tcc gtc gca ctg gca cca cac 1475

Thr Gly Glu His Arg Arg Asp Lys Arg Ser Val Ala Leu Ala Pro His

100 105 110

gta ggg ctt ggt cta gaa aca aga acc gaa acg tgg atg tcc tct gaa 1523

Val Gly Leu Gly Leu Glu Thr Arg Thr Glu Thr Trp Met Ser Ser Glu

115 120 125

ggc gct tgg aaa caa ata caa aaa gtg gag acc tgg gct ctg aga cac 1571

Gly Ala Trp Lys Gln Ile Gln Lys Val Glu Thr Trp Ala Leu Arg His

130 135 140 145

cca gga ttc acg gtg ata gcc ctt ttt cta gca cat gcc ata gga aca 1619

Pro Gly Phe Thr Val Ile Ala Leu Phe Leu Ala His Ala Ile Gly Thr

150 155 160

tcc atc acc cag aaa ggg atc att ttt att ttg ctg atg ctg gta act 1667

Ser Ile Thr Gln Lys Gly Ile Ile Phe Ile Leu Leu Met Leu Val Thr

165 170 175

cca tcc atg gcc atg cgg tgc gtg gga ata ggc aac aga gac ttc gtg 1715

Pro Ser Met Ala Met Arg Cys Val Gly Ile Gly Asn Arg Asp Phe Val

180 185 190

gaa gga ctg tca gga gct acg tgg gtg gat gtg gta ctg gag cat gga 1763

Glu Gly Leu Ser Gly Ala Thr Trp Val Asp Val Val Leu Glu His Gly

195 200 205

agt tgc gtc act acc atg gca aaa gac aaa cca aca ctg gac att gaa 1811

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

210 215 220 225

ctc ttg aag acg gag gtc aca aac cct gcc gtc ctg cgc aaa ctg tgc 1859

Leu Leu Lys Thr Glu Val Thr Asn Pro Ala Val Leu Arg Lys Leu Cys

230 235 240

att gaa gct aaa ata tca aac acc acc acc gat tcg aga tgt cca aca 1907

Ile Glu Ala Lys Ile Ser Asn Thr Thr Thr Asp Ser Arg Cys Pro Thr

245 250 255

caa gga gaa gcc acg ctg gtg gaa gaa cag gac acg aac ttt gtg tgt 1955

Gln Gly Glu Ala Thr Leu Val Glu Glu Gln Asp Thr Asn Phe Val Cys

260 265 270

cga cga acg ttc gtg gac aga ggc tgg ggc aat ggt tgt ggg cta gcc 2003

Arg Arg Thr Phe Val Asp Arg Gly Trp Gly Asn Gly Cys Gly Leu Ala

275 280 285

gga aaa ggt agc tta ata acg tgt gct aag ttt aag tgt gtg aca aaa 2051

Gly Lys Gly Ser Leu Ile Thr Cys Ala Lys Phe Lys Cys Val Thr Lys

290 295 300 305

ctg gaa gga aag ata gtc caa tat gaa aac tta aaa tat tca gtg ata 2099

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

310 315 320

gtc acc gta cac act gga gac cag cac caa gtt gga aat gag acc aca 2147

Val Thr Val His Thr Gly Asp Gln His Gln Val Gly Asn Glu Thr Thr

325 330 335

gaa cat gga aca act gca acc ata aca cct caa gct ccc acg tcg gaa 2195

Glu His Gly Thr Thr Ala Thr Ile Thr Pro Gln Ala Pro Thr Ser Glu

340 345 350

ata cag ctg aca gac tac gga gct cta aca ttg gat tgt tca cct aga 2243

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

355 360 365

aca ggg cta gac ttt aat gag atg gtg ttg ttg aca atg gaa aaa aaa 2291

Thr Gly Leu Asp Phe Asn Glu Met Val Leu Leu Thr Met Glu Lys Lys

370 375 380 385

tca tgg ctc gtc cac aaa caa tgg ttt cta gac tta cca ctg cct tgg 2339

Ser Trp Leu Val His Lys Gln Trp Phe Leu Asp Leu Pro Leu Pro Trp

390 395 400

acc tcg ggg gct tca aca tcc caa gag act tgg aat aga caa gac ttg 2387

Thr Ser Gly Ala Ser Thr Ser Gln Glu Thr Trp Asn Arg Gln Asp Leu

405 410 415

ctg gtc aca ttt aag aca gct cat gca atg aag cag gaa gta gtc gta 2435

Leu Val Thr Phe Lys Thr Ala His Ala Met Lys Gln Glu Val Val Val

420 425 430

cta gga tca caa gaa gga gca atg cac act gcg ttg act gga gcg aca 2483

Leu Gly Ser Gln Glu Gly Ala Met His Thr Ala Leu Thr Gly Ala Thr

435 440 445

gaa atc caa acg tct gga acg aca aca att ttt gca gga cac ctg aaa 2531

Glu Ile Gln Thr Ser Gly Thr Thr Thr Ile Phe Ala Gly His Leu Lys

450 455 460 465

tgc aga cta aaa atg gat aaa ctg act tta aaa ggg atg tca tat gta 2579

Cys Arg Leu Lys Met Asp Lys Leu Thr Leu Lys Gly Met Ser Tyr Val

470 475 480

atg tgc aca ggg tca ttc aag tta gag aag gaa gtg gct gag acc cag 2627

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

485 490 495

cat gga act gtt cta gtg cag gtt aaa tac gaa gga aca gat gca cca 2675

His Gly Thr Val Leu Val Gln Val Lys Tyr Glu Gly Thr Asp Ala Pro

500 505 510

tgc aag atc ccc ttc tcg tcc caa gat gag aag gga gta acc cag aat 2723

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

515 520 525

ggg aga ttg ata aca gcc aac ccc ata gtc act gac aaa gaa aaa cca 2771

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

530 535 540 545

gtc aac att gaa gcg gag cca cct ttt ggt gag agc tac att gtg gta 2819

Val Asn Ile Glu Ala Glu Pro Pro Phe Gly Glu Ser Tyr Ile Val Val

550 555 560

gga gca ggt gaa aaa gct ttg aaa cta agc tgg ttc aag aag gga agc 2867

Gly Ala Gly Glu Lys Ala Leu Lys Leu Ser Trp Phe Lys Lys Gly Ser

565 570 575

agt ata ggg aaa atg ttt gaa gca act gcc cgt gga gca cga agg atg 2915

Ser Ile Gly Lys Met Phe Glu Ala Thr Ala Arg Gly Ala Arg Arg Met

580 585 590

gcc atc ctg gga gac act gca tgg gac ttc ggt tct ata gga ggg gtg 2963

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

595 600 605

ttc acg tct gtg gga aaa ctg ata cac cag att ttt ggg act gcg tat 3011

Phe Thr Ser Val Gly Lys Leu Ile His Gln Ile Phe Gly Thr Ala Tyr

610 615 620 625

gga gtt ttg ttc agc ggt gtt tct tgg acc atg aag ata gga ata ggg 3059

Gly Val Leu Phe Ser Gly Val Ser Trp Thr Met Lys Ile Gly Ile Gly

630 635 640

att ctg ctg aca tgg cta gga tta aac tca agg agc acg tcc ctt tca 3107

Ile Leu Leu Thr Trp Leu Gly Leu Asn Ser Arg Ser Thr Ser Leu Ser

645 650 655

atg acg tgt atc gca gtt ggc atg gtc acg ctg tac cta gga gtc atg 3155

Met Thr Cys Ile Ala Val Gly Met Val Thr Leu Tyr Leu Gly Val Met

660 665 670

gtt cag gcg taataaagat ctgctgtgcc ttctagttgc cagccatctg 3204

Val Gln Ala

675

ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt 3264

cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg 3324

gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg 3384

atgcggtggg ctctatgggt acccaggtgc tgaagaattg acccggttcc tcctgggcca 3444

gaaagaagca ggcacatccc cttctctgtg acacaccctg tccacgcccc tggttcttag 3504

ttccagcccc actcatagga cactcatagc tcaggagggc tccgccttca atcccacccg 3564

ctaaagtact tggagcggtc tctccctccc tcatcagccc accaaaccaa acctagcctc 3624

caagagtggg aagaaattaa agcaagatag gctattaagt gcagagggag agaaaatgcc 3684

tccaacatgt gaggaagtaa tgagagaaat catagaattt taaggccatg atttaaggcc 3744

atcatggcct aagcttgaaa ggagatagga tcaaagcttg gcgtaatcat ggtcatagct 3804

gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 3864

aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 3924

actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 3984

cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 4044

gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 4104

atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 4164

caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 4224

gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 4284

ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 4344

cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 4404

taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 4464

cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 4524

acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 4584

aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 4644

atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 4704

atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 4764

gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 4824

gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 4884

ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 4944

ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 5004

tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 5064

accatctggc cccagtgctg caatgatacc gcgagaacca cgctcaccgg ctccagattt 5124

atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 5184

cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 5244

tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 5304

tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 5364

gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 5424

agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 5484

aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 5544

gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 5604

tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 5664

gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 5724

tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 5784

aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 5844

catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 5904

acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat 5964

tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gggtcgcgcg 6024

tttcggtgat gacggtgaaa acctctgaca catgcagctc ccgttgacgg tcacagcttg 6084

tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg 6144

gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccataa 6204

aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt 6264

ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agcccgagat 6324

agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa 6384

cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccaa 6444

atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc 6504

ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc 6564

gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac 6624

acccgccgcg cttaatgcgc cgctacaggg cgcgtactat ggttgctttg acgtatgcgg 6684

tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc 6744

aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg 6804

gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca 6864

cgacgttgta aaacgacggc cagtgaattc catggtctca actttc 6910

<210> 16

<211> 676

<212> PRT

<213> 登革病毒

<400> 16

Met Ser Val Thr Met Leu Leu Met Leu Leu Pro Thr Ala Leu Ala Phe

1 5 10 15

His Leu Thr Thr Arg Gly Gly Glu Pro His Met Ile Val Ser Lys Gln

20 25 30

Glu Arg Gly Lys Ser Leu Leu Phe Lys Thr Ser Ala Gly Val Asn Met

35 40 45

Cys Thr Leu Ile Ala Met Asp Leu Gly Glu Leu Cys Glu Asp Thr Met

50 55 60

Thr Tyr Lys Cys Pro Arg Ile Thr Glu Thr Glu Pro Asp Asp Val Asp

65 70 75 80

Cys Trp Cys Asn Ala Thr Glu Thr Trp Val Thr Tyr Gly Thr Cys Ser

85 90 95

Gln Thr Gly Glu His Arg Arg Asp Lys Arg Ser Val Ala Leu Ala Pro

100 105 110

His Val Gly Leu Gly Leu Glu Thr Arg Thr Glu Thr Trp Met Ser Ser

115 120 125

Glu Gly Ala Trp Lys Gln Ile Gln Lys Val Glu Thr Trp Ala Leu Arg

130 135 140

His Pro Gly Phe Thr Val Ile Ala Leu Phe Leu Ala His Ala Ile Gly

145 150 155 160

Thr Ser Ile Thr Gln Lys Gly Ile Ile Phe Ile Leu Leu Met Leu Val

165 170 175

Thr Pro Ser Met Ala Met Arg Cys Val Gly Ile Gly Asn Arg Asp Phe

180 185 190

Val Glu Gly Leu Ser Gly Ala Thr Trp Val Asp Val Val Leu Glu His

195 200 205

Gly Ser Cys Val Thr Thr Met Ala Lys Asp Lys Pro Thr Leu Asp Ile

210 215 220

Glu Leu Leu Lys Thr Glu Val Thr Asn Pro Ala Val Leu Arg Lys Leu

225 230 235 240

Cys Ile Glu Ala Lys Ile Ser Asn Thr Thr Thr Asp Ser Arg Cys Pro

245 250 255

Thr Gln Gly Glu Ala Thr Leu Val Glu Glu Gln Asp Thr Asn Phe Val

260 265 270

Cys Arg Arg Thr Phe Val Asp Arg Gly Trp Gly Asn Gly Cys Gly Leu

275 280 285

Ala Gly Lys Gly Ser Leu Ile Thr Cys Ala Lys Phe Lys Cys Val Thr

290 295 300

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

305 310 315 320

Ile Val Thr Val His Thr Gly Asp Gln His Gln Val Gly Asn Glu Thr

325 330 335

Thr Glu His Gly Thr Thr Ala Thr Ile Thr Pro Gln Ala Pro Thr Ser

340 345 350

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

355 360 365

Arg Thr Gly Leu Asp Phe Asn Glu Met Val Leu Leu Thr Met Glu Lys

370 375 380

Lys Ser Trp Leu Val His Lys Gln Trp Phe Leu Asp Leu Pro Leu Pro

385 390 395 400

Trp Thr Ser Gly Ala Ser Thr Ser Gln Glu Thr Trp Asn Arg Gln Asp

405 410 415

Leu Leu Val Thr Phe Lys Thr Ala His Ala Met Lys Gln Glu Val Val

420 425 430

Val Leu Gly Ser Gln Glu Gly Ala Met His Thr Ala Leu Thr Gly Ala

435 440 445

Thr Glu Ile Gln Thr Ser Gly Thr Thr Thr Ile Phe Ala Gly His Leu

450 455 460

Lys Cys Arg Leu Lys Met Asp Lys Leu Thr Leu Lys Gly Met Ser Tyr

465 470 475 480

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

485 490 495

Gln His Gly Thr Val Leu Val Gln Val Lys Tyr Glu Gly Thr Asp Ala

500 505 510

Pro Cys Lys Ile Pro Phe Ser Ser Gln Asp Glu Lys Gly Val Thr Gln

515 520 525

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

530 535 540

Pro Val Asn Ile Glu Ala Glu Pro Pro Phe Gly Glu Ser Tyr Ile Val

545 550 555 560

Val Gly Ala Gly Glu Lys Ala Leu Lys Leu Ser Trp Phe Lys Lys Gly

565 570 575

Ser Ser Ile Gly Lys Met Phe Glu Ala Thr Ala Arg Gly Ala Arg Arg

580 585 590

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

595 600 605

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

610 615 620

Tyr Gly Val Leu Phe Ser Gly Val Ser Trp Thr Met Lys Ile Gly Ile

625 630 635 640

Gly Ile Leu Leu Thr Trp Leu Gly Leu Asn Ser Arg Ser Thr Ser Leu

645 650 655

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

660 665 670

Met Val Gln Ala

675

<210> 17

<211> 14

<212> PRT

<213> 登革病毒

<220>

<221> 信号

<222> (1)..(14)

<400> 17

Ser Val Thr Met Leu Leu Met Leu Leu Pro Thr Ala Leu Ala

1 5 10

<210> 18

<211> 91

<212> PRT

<213> 登革病毒

<220>

<221> misc_feature

<222> (1)..(91)

<223> pr序列

<220>

<221> misc_feature

<222> (1)..(91)

<223> DENV1 pr序列

<400> 18

Phe His Leu Thr Thr Arg Gly Gly Glu Pro His Met Ile Val Ser Lys

1 5 10 15

Gln Glu Arg Gly Lys Ser Leu Leu Phe Lys Thr Ser Ala Gly Val Asn

20 25 30

Met Cys Thr Leu Ile Ala Met Asp Leu Gly Glu Leu Cys Glu Asp Thr

35 40 45

Met Thr Tyr Lys Cys Pro Arg Ile Thr Glu Thr Glu Pro Asp Asp Val

50 55 60

Asp Cys Trp Cys Asn Ala Thr Glu Thr Trp Val Thr Tyr Gly Thr Cys

65 70 75 80

Ser Gln Thr Gly Glu His Arg Arg Asp Lys Arg

85 90

<210> 19

<211> 75

<212> PRT

<213> 登革病毒

<220>

<221> MISC_FEATURE

<222> (1)..(75)

<223> M序列

<220>

<221> MISC_FEATURE

<222> (1)..(75)

<223> DENV1 M序列

<400> 19

Ser Val Ala Leu Ala Pro His Val Gly Leu Gly Leu Glu Thr Arg Thr

1 5 10 15

Glu Thr Trp Met Ser Ser Glu Gly Ala Trp Lys Gln Ile Gln Lys Val

20 25 30

Glu Thr Trp Ala Leu Arg His Pro Gly Phe Thr Val Ile Ala Leu Phe

35 40 45

Leu Ala His Ala Ile Gly Thr Ser Ile Thr Gln Lys Gly Ile Ile Phe

50 55 60

Ile Leu Leu Met Leu Val Thr Pro Ser Met Ala

65 70 75

<210> 20

<211> 495

<212> PRT

<213> 登革病毒

<220>

<221> MISC_FEATURE

<222> (1)..(495)

<223> DENV1包膜序列

<400> 20

Met Arg Cys Val Gly Ile Gly Asn Arg Asp Phe Val Glu Gly Leu Ser

1 5 10 15

Gly Ala Thr Trp Val Asp Val Val Leu Glu His Gly Ser Cys Val Thr

20 25 30

Thr Met Ala Lys Asp Lys Pro Thr Leu Asp Ile Glu Leu Leu Lys Thr

35 40 45

Glu Val Thr Asn Pro Ala Val Leu Arg Lys Leu Cys Ile Glu Ala Lys

50 55 60

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

65 70 75 80

Thr Leu Val Glu Glu Gln Asp Thr Asn Phe Val Cys Arg Arg Thr Phe

85 90 95

Val Asp Arg Gly Trp Gly Asn Gly Cys Gly Leu Phe Gly Lys Gly Ser

100 105 110

Leu Ile Thr Cys Ala Lys Phe Lys Cys Val Thr Lys Leu Glu Gly Lys

115 120 125

Ile Val Gln Tyr Glu Asn Leu Lys Tyr Ser Val Ile Val Thr Val His

130 135 140

Thr Gly Asp Gln His Gln Val Gly Asn Glu Thr Thr Glu His Gly Thr

145 150 155 160

Thr Ala Thr Ile Thr Pro Gln Ala Pro Thr Ser Glu Ile Gln Leu Thr

165 170 175

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

180 185 190

Phe Asn Glu Met Val Leu Leu Thr Met Glu Lys Lys Ser Trp Leu Val

195 200 205

His Lys Gln Trp Phe Leu Asp Leu Pro Leu Pro Trp Thr Ser Gly Ala

210 215 220

Ser Thr Ser Gln Glu Thr Trp Asn Arg Gln Asp Leu Leu Val Thr Phe

225 230 235 240

Lys Thr Ala His Ala Lys Lys Gln Glu Val Val Val Leu Gly Ser Gln

245 250 255

Glu Gly Ala Met His Thr Ala Leu Thr Gly Ala Thr Glu Ile Gln Thr

260 265 270

Ser Gly Thr Thr Thr Ile Phe Ala Gly His Leu Lys Cys Arg Leu Lys

275 280 285

Met Asp Lys Leu Thr Leu Lys Gly Met Ser Tyr Val Met Cys Thr Gly

290 295 300

Ser Phe Lys Leu Glu Lys Glu Val Ala Glu Thr Gln His Gly Thr Val

305 310 315 320

Leu Val Gln Val Lys Tyr Glu Gly Thr Asp Ala Pro Cys Lys Ile Pro

325 330 335

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

340 345 350

Thr Ala Asn Pro Ile Val Thr Asp Lys Glu Lys Pro Val Asn Ile Glu

355 360 365

Ala Glu Pro Pro Phe Gly Glu Ser Tyr Ile Val Val Gly Ala Gly Glu

370 375 380

Lys Ala Leu Lys Leu Ser Trp Phe Lys Lys Gly Ser Ser Ile Gly Lys

385 390 395 400

Met Phe Glu Ala Thr Ala Arg Gly Ala Arg Arg Met Ala Ile Leu Gly

405 410 415

Asp Thr Ala Trp Asp Phe Gly Ser Ile Gly Gly Val Phe Thr Ser Val

420 425 430

Gly Lys Leu Ile His Gln Ile Phe Gly Thr Ala Tyr Gly Val Leu Phe

435 440 445

Ser Gly Val Ser Trp Thr Met Lys Ile Gly Ile Gly Ile Leu Leu Thr

450 455 460

Trp Leu Gly Leu Asn Ser Arg Ser Thr Ser Leu Ser Met Thr Cys Ile

465 470 475 480

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

485 490 495

<210> 21

<211> 676

<212> PRT

<213> 登革病毒

<220>

<221> INIT_MET

<222> (1)..(1)

<220>

<221> 信号

<222> (2)..(15)

<220>

<221> MISC_FEATURE

<222> (16)..(106)

<220>

<221> MISC_FEATURE

<222> (107)..(181)

<223> DENV1 M序列

<220>

<221> MISC_FEATURE

<222> (182)..(676)

<223> DENV1包膜序列

<400> 21

Met Ser Val Thr Met Leu Leu Met Leu Leu Pro Thr Ala Leu Ala Phe

1 5 10 15

His Leu Thr Thr Arg Gly Gly Glu Pro His Met Ile Val Ser Lys Gln

20 25 30

Glu Arg Gly Lys Ser Leu Leu Phe Lys Thr Ser Ala Gly Val Asn Met

35 40 45

Cys Thr Leu Ile Ala Met Asp Leu Gly Glu Leu Cys Glu Asp Thr Met

50 55 60

Thr Tyr Lys Cys Pro Arg Ile Thr Glu Thr Glu Pro Asp Asp Val Asp

65 70 75 80

Cys Trp Cys Asn Ala Thr Glu Thr Trp Val Thr Tyr Gly Thr Cys Ser

85 90 95

Gln Thr Gly Glu His Arg Arg Asp Lys Arg Ser Val Ala Leu Ala Pro

100 105 110

His Val Gly Leu Gly Leu Glu Thr Arg Thr Glu Thr Trp Met Ser Ser

115 120 125

Glu Gly Ala Trp Lys Gln Ile Gln Lys Val Glu Thr Trp Ala Leu Arg

130 135 140

His Pro Gly Phe Thr Val Ile Ala Leu Phe Leu Ala His Ala Ile Gly

145 150 155 160

Thr Ser Ile Thr Gln Lys Gly Ile Ile Phe Ile Leu Leu Met Leu Val

165 170 175

Thr Pro Ser Met Ala Met Arg Cys Val Gly Ile Gly Asn Arg Asp Phe

180 185 190

Val Glu Gly Leu Ser Gly Ala Thr Trp Val Asp Val Val Leu Glu His

195 200 205

Gly Ser Cys Val Thr Thr Met Ala Lys Asp Lys Pro Thr Leu Asp Ile

210 215 220

Glu Leu Leu Lys Thr Glu Val Thr Asn Pro Ala Val Leu Arg Lys Leu

225 230 235 240

Cys Ile Glu Ala Lys Ile Ser Asn Thr Thr Thr Asp Ser Arg Cys Pro

245 250 255

Thr Gln Gly Glu Ala Thr Leu Val Glu Glu Gln Asp Thr Asn Phe Val

260 265 270

Cys Arg Arg Thr Phe Val Asp Arg Gly Trp Gly Asn Gly Cys Gly Leu

275 280 285

Phe Gly Lys Gly Ser Leu Ile Thr Cys Ala Lys Phe Lys Cys Val Thr

290 295 300

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

305 310 315 320

Ile Val Thr Val His Thr Gly Asp Gln His Gln Val Gly Asn Glu Thr

325 330 335

Thr Glu His Gly Thr Thr Ala Thr Ile Thr Pro Gln Ala Pro Thr Ser

340 345 350

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

355 360 365

Arg Thr Gly Leu Asp Phe Asn Glu Met Val Leu Leu Thr Met Glu Lys

370 375 380

Lys Ser Trp Leu Val His Lys Gln Trp Phe Leu Asp Leu Pro Leu Pro

385 390 395 400

Trp Thr Ser Gly Ala Ser Thr Ser Gln Glu Thr Trp Asn Arg Gln Asp

405 410 415

Leu Leu Val Thr Phe Lys Thr Ala His Ala Lys Lys Gln Glu Val Val

420 425 430

Val Leu Gly Ser Gln Glu Gly Ala Met His Thr Ala Leu Thr Gly Ala

435 440 445

Thr Glu Ile Gln Thr Ser Gly Thr Thr Thr Ile Phe Ala Gly His Leu

450 455 460

Lys Cys Arg Leu Lys Met Asp Lys Leu Thr Leu Lys Gly Met Ser Tyr

465 470 475 480

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

485 490 495

Gln His Gly Thr Val Leu Val Gln Val Lys Tyr Glu Gly Thr Asp Ala

500 505 510

Pro Cys Lys Ile Pro Phe Ser Ser Gln Asp Glu Lys Gly Val Thr Gln

515 520 525

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

530 535 540

Pro Val Asn Ile Glu Ala Glu Pro Pro Phe Gly Glu Ser Tyr Ile Val

545 550 555 560

Val Gly Ala Gly Glu Lys Ala Leu Lys Leu Ser Trp Phe Lys Lys Gly

565 570 575

Ser Ser Ile Gly Lys Met Phe Glu Ala Thr Ala Arg Gly Ala Arg Arg

580 585 590

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

595 600 605

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

610 615 620

Tyr Gly Val Leu Phe Ser Gly Val Ser Trp Thr Met Lys Ile Gly Ile

625 630 635 640

Gly Ile Leu Leu Thr Trp Leu Gly Leu Asn Ser Arg Ser Thr Ser Leu

645 650 655

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

660 665 670

Met Val Gln Ala

675

<210> 22

<211> 676

<212> PRT

<213> 登革病毒

<220>

<221> MISC_FEATURE

<223> DEN2 prM E (F108A)_3' DEN1

<220>

<221> MISC_FEATURE

<222> (182)..(477)

<223> DEN2 prM E (F108A)

<220>

<221> MISC_FEATURE

<222> (182)..(477)

<223> DEV2 prM E (F108A)

<220>

<221> MISC_FEATURE

<222> (478)..(676)

<223> 3' DEV1

<400> 22

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

1 5 10 15

His Leu Thr Thr Arg Asn Gly Glu Pro His Met Ile Val Ser Arg Gln

20 25 30

Glu Lys Gly Lys Ser Leu Leu Phe Lys Thr Lys Asp Gly Thr Asn Met

35 40 45

Cys Thr Leu Met Ala Met Asp Leu Gly Glu Leu Cys Glu Asp Thr Ile

50 55 60

Thr Tyr Lys Cys Pro Phe Leu Lys Gln Asn Glu Pro Glu Asp Ile Asp

65 70 75 80

Cys Trp Cys Asn Ser Thr Ser Thr Trp Val Thr Tyr Gly Thr Cys Thr

85 90 95

Thr Thr Gly Glu His Arg Arg Glu Lys Arg Ser Val Ala Leu Val Pro

100 105 110

His Val Gly Met Gly Leu Glu Thr Arg Thr Glu Thr Trp Met Ser Ser

115 120 125

Glu Gly Ala Trp Lys His Ala Gln Arg Ile Glu Thr Trp Ile Leu Arg

130 135 140

His Pro Gly Phe Thr Ile Met Ala Ala Ile Leu Ala Tyr Thr Ile Gly

145 150 155 160

Thr Thr His Phe Gln Arg Val Leu Ile Phe Ile Leu Leu Thr Ala Ile

165 170 175

Ala Pro Ser Met Thr Met Arg Cys Ile Gly Ile Ser Asn Arg Asp Phe

180 185 190

Val Glu Gly Val Ser Gly Gly Ser Trp Val Asp Ile Val Leu Glu His

195 200 205

Gly Ser Cys Val Thr Thr Met Ala Lys Asn Lys Pro Thr Leu Asp Phe

210 215 220

Glu Leu Ile Lys Thr Glu Ala Lys Gln Pro Ala Thr Leu Arg Lys Tyr

225 230 235 240

Cys Ile Glu Ala Lys Leu Thr Asn Thr Thr Thr Asp Ser Arg Cys Pro

245 250 255

Thr Gln Gly Glu Pro Thr Leu Asn Glu Glu Gln Asp Lys Arg Phe Val

260 265 270

Cys Lys His Ser Met Val Asp Arg Gly Trp Gly Asn Gly Cys Gly Leu

275 280 285

Ala Gly Lys Gly Gly Ile Val Thr Cys Ala Met Phe Thr Cys Lys Lys

290 295 300

Asn Met Glu Gly Lys Ile Val Gln Pro Glu Asn Leu Glu Tyr Thr Val

305 310 315 320

Val Ile Thr Pro His Ser Gly Glu Glu His Ala Val Gly Asn Asp Thr

325 330 335

Gly Lys His Gly Lys Glu Val Lys Ile Thr Pro Gln Ser Ser Ile Thr

340 345 350

Glu Ala Glu Leu Thr Gly Tyr Gly Thr Val Thr Met Glu Cys Ser Pro

355 360 365

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

370 375 380

Lys Ala Trp Leu Val His Arg Gln Trp Phe Leu Asp Leu Pro Leu Pro

385 390 395 400

Trp Leu Pro Gly Ala Asp Thr Gln Gly Ser Asn Trp Ile Gln Lys Glu

405 410 415

Thr Leu Val Thr Phe Lys Asn Pro His Ala Lys Lys Gln Asp Val Val

420 425 430

Val Leu Gly Ser Gln Glu Gly Ala Met His Thr Ala Leu Thr Gly Ala

435 440 445

Thr Glu Ile Gln Met Ser Ser Gly Asn Leu Leu Phe Thr Gly His Leu

450 455 460

Lys Cys Arg Leu Arg Met Asp Lys Leu Gln Leu Lys Gly Met Ser Tyr

465 470 475 480

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

485 490 495

Gln His Gly Thr Val Leu Val Gln Val Lys Tyr Glu Gly Thr Asp Ala

500 505 510

Pro Cys Lys Ile Pro Phe Ser Ser Gln Asp Glu Lys Gly Val Thr Gln

515 520 525

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

530 535 540

Pro Val Asn Ile Glu Ala Glu Pro Pro Phe Gly Glu Ser Tyr Ile Val

545 550 555 560

Val Gly Ala Gly Glu Lys Ala Leu Lys Leu Ser Trp Phe Lys Lys Gly

565 570 575

Ser Ser Ile Gly Lys Met Phe Glu Ala Thr Ala Arg Gly Ala Arg Arg

580 585 590

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

595 600 605

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

610 615 620

Tyr Gly Val Leu Phe Ser Gly Val Ser Trp Thr Met Lys Ile Gly Ile

625 630 635 640

Gly Ile Leu Leu Thr Trp Leu Gly Leu Asn Ser Arg Ser Thr Ser Leu

645 650 655

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

660 665 670

Met Val Gln Ala

675

<210> 23

<211> 674

<212> PRT

<213> 登革病毒

<220>

<221> misc_feature

<223> DEV3 prM E (F108A)_3'DEV1

<220>

<221> misc_feature

<222> (182)..(475)

<223> DEV3 prM E (F108A)

<220>

<221> misc_feature

<222> (476)..(676)

<223> 3' DEV1

<400> 23

Met Thr Ser Leu Cys Leu Met Met Ile Leu Pro Ala Thr Leu Ala Phe

1 5 10 15

His Leu Thr Ser Arg Asp Gly Glu Pro Arg Met Ile Val Gly Lys Asn

20 25 30

Glu Arg Gly Lys Ser Leu Leu Phe Lys Thr Ala Ser Gly Ile Asn Met

35 40 45

Cys Thr Leu Ile Ala Met Asp Leu Gly Glu Met Cys Asp Asp Thr Val

50 55 60

Thr Tyr Lys Cys Pro Leu Ile Ala Glu Val Glu Pro Glu Asp Ile Asp

65 70 75 80

Cys Trp Cys Asn Leu Thr Ser Thr Trp Val Thr Tyr Gly Thr Cys Asn

85 90 95

Gln Ala Gly Glu His Arg Arg Asp Lys Arg Ser Val Ala Leu Ala Pro

100 105 110

His Val Gly Met Gly Leu Asp Thr Arg Thr Gln Thr Trp Met Ser Ala

115 120 125

Glu Gly Ala Trp Arg Gln Val Glu Lys Val Glu Thr Trp Ala Leu Arg

130 135 140

His Pro Gly Phe Thr Ile Leu Ala Leu Phe Leu Ala His Tyr Ile Gly

145 150 155 160

Thr Ser Leu Thr Gln Lys Val Val Ile Phe Ile Leu Leu Met Leu Val

165 170 175

Thr Pro Ser Met Thr Met Arg Cys Val Gly Val Gly Asn Arg Asp Phe

180 185 190

Val Glu Gly Leu Ser Gly Ala Thr Trp Val Asp Val Val Leu Glu His

195 200 205

Gly Gly Cys Val Thr Thr Met Ala Lys Asn Lys Pro Thr Leu Asp Ile

210 215 220

Glu Leu Gln Lys Thr Glu Ala Thr Gln Leu Ala Thr Leu Arg Lys Leu

225 230 235 240

Cys Ile Glu Gly Lys Ile Thr Asn Ile Thr Thr Asp Ser Arg Cys Pro

245 250 255

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

260 265 270

Cys Lys His Thr Tyr Val Asp Arg Gly Trp Gly Asn Gly Cys Gly Leu

275 280 285

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

290 295 300

Pro Ile Glu Gly Lys Val Val Gln His Glu Asn Leu Lys Tyr Thr Val

305 310 315 320

Ile Ile Thr Val His Thr Gly Asp Gln His Gln Val Gly Asn Asp Thr

325 330 335

Gln Gly Val Thr Val Glu Ile Thr Pro Gln Ala Ser Thr Val Glu Ala

340 345 350

Ile Leu Pro Glu Tyr Gly Thr Leu Gly Leu Glu Cys Ser Pro Arg Thr

355 360 365

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

370 375 380

Trp Met Val His Arg Gln Trp Phe Phe Asp Leu Pro Leu Pro Trp Thr

385 390 395 400

Ser Gly Ala Thr Thr Glu Ala Pro Thr Trp Asn Arg Lys Glu Leu Leu

405 410 415

Val Thr Phe Lys Asn Ala His Ala Lys Lys Gln Glu Val Val Val Leu

420 425 430

Gly Ser Gln Glu Gly Ala Met His Thr Ala Leu Thr Gly Ala Thr Glu

435 440 445

Ile Gln Asn Ser Gly Gly Thr Ser Ile Phe Ala Gly His Leu Lys Cys

450 455 460

Arg Leu Lys Met Asp Lys Leu Glu Leu Lys Gly Met Ser Tyr Val Met

465 470 475 480

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

485 490 495

Gly Thr Val Leu Val Gln Val Lys Tyr Glu Gly Thr Asp Ala Pro Cys

500 505 510

Lys Ile Pro Phe Ser Ser Gln Asp Glu Lys Gly Val Thr Gln Asn Gly

515 520 525

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

530 535 540

Asn Ile Glu Ala Glu Pro Pro Phe Gly Glu Ser Tyr Ile Val Val Gly

545 550 555 560

Ala Gly Glu Lys Ala Leu Lys Leu Ser Trp Phe Lys Lys Gly Ser Ser

565 570 575

Ile Gly Lys Met Phe Glu Ala Thr Ala Arg Gly Ala Arg Arg Met Ala

580 585 590

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

595 600 605

Thr Ser Val Gly Lys Leu Ile His Gln Ile Phe Gly Thr Ala Tyr Gly

610 615 620

Val Leu Phe Ser Gly Val Ser Trp Thr Met Lys Ile Gly Ile Gly Ile

625 630 635 640

Leu Leu Thr Trp Leu Gly Leu Asn Ser Arg Ser Thr Ser Leu Ser Met

645 650 655

Thr Cys Ile Ala Val Gly Met Val Thr Leu Tyr Leu Gly Val Met Val

660 665 670

Gln Ala

<210> 24

<211> 676

<212> PRT

<213> 登革病毒

<220>

<221> MISC_FEATURE

<223> DEV4 prM E (F108A)_3'DEV1

<220>

<221> MISC_FEATURE

<222> (1)..(477)

<223> DEV4 prM E (F108A)

<220>

<221> MISC_FEATURE

<222> (478)..(676)

<223> 3' DEV1

<400> 24

Met Ser Thr Val Thr Leu Leu Cys Leu Ile Pro Thr Val Met Ala Phe

1 5 10 15

His Leu Ser Thr Arg Asp Gly Glu Pro Leu Met Ile Val Ala Lys His

20 25 30

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

35 40 45

Cys Thr Leu Ile Ala Met Asp Leu Gly Glu Met Cys Glu Asp Thr Val

50 55 60

Thr Tyr Lys Cys Pro Leu Leu Val Asn Thr Glu Pro Glu Asp Ile Asp

65 70 75 80

Cys Trp Cys Asn Leu Thr Ser Thr Trp Val Met Tyr Gly Thr Cys Thr

85 90 95

Gln Asn Gly Glu Arg Arg Arg Glu Lys Arg Ser Val Ala Leu Thr Pro

100 105 110

His Ser Gly Met Gly Leu Glu Thr Arg Ala Glu Thr Trp Met Ser Ser

115 120 125

Glu Gly Ala Trp Lys His Ala Gln Arg Val Glu Ser Trp Ile Leu Arg

130 135 140

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

145 150 155 160

Gln Thr Gly Ile Gln Arg Thr Val Phe Phe Val Leu Met Met Leu Val

165 170 175

Ala Pro Ser Tyr Gly Met Arg Cys Ile Gly Val Gly Asn Arg Asp Phe

180 185 190

Val Glu Gly Val Ser Gly Gly Ala Trp Val Asp Leu Val Leu Glu His

195 200 205

Gly Gly Cys Val Thr Thr Met Ala Gln Gly Lys Pro Thr Leu Asp Phe

210 215 220

Glu Leu Ile Lys Thr Thr Ala Lys Glu Val Ala Leu Leu Arg Thr Tyr

225 230 235 240

Cys Ile Glu Ala Ser Ile Ser Asn Ile Thr Thr Ala Thr Arg Cys Pro

245 250 255

Thr Gln Gly Glu Pro Tyr Leu Lys Glu Glu Gln Asp Gln Gln Tyr Ile

260 265 270

Cys Arg Arg Asp Val Val Asp Arg Gly Trp Gly Asn Gly Cys Gly Leu

275 280 285

Ala Gly Lys Gly Gly Val Val Thr Cys Ala Lys Phe Ser Cys Ser Gly

290 295 300

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

305 310 315 320

Val Val Thr Val His Asn Gly Asp Thr His Ala Val Gly Asn Asp Thr

325 330 335

Ser Asn His Gly Val Thr Ala Thr Ile Thr Pro Arg Ser Pro Ser Val

340 345 350

Glu Val Glu Leu Pro Asp Tyr Gly Glu Leu Ser Leu Asp Cys Glu Pro

355 360 365

Arg Ser Gly Ile Asp Phe Asn Glu Met Ile Leu Met Lys Met Glu Lys

370 375 380

Lys Thr Trp Leu Val His Lys Gln Trp Phe Leu Asp Leu Pro Leu Pro

385 390 395 400

Trp Thr Ala Gly Ala Asp Thr Ser Glu Val His Trp Asn His Lys Glu

405 410 415

Arg Met Val Thr Phe Lys Val Pro His Ala Lys Arg Gln Asp Val Thr

420 425 430

Val Leu Gly Ser Gln Glu Gly Ala Met His Ser Ala Leu Thr Gly Ala

435 440 445

Thr Glu Val Asp Ser Gly Asp Gly Asn His Met Phe Ala Gly His Leu

450 455 460

Lys Cys Lys Val Arg Met Glu Lys Leu Arg Ile Lys Gly Met Ser Tyr

465 470 475 480

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

485 490 495

Gln His Gly Thr Val Leu Val Gln Val Lys Tyr Glu Gly Thr Asp Ala

500 505 510

Pro Cys Lys Ile Pro Phe Ser Ser Gln Asp Glu Lys Gly Val Thr Gln

515 520 525

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

530 535 540

Pro Val Asn Ile Glu Ala Glu Pro Pro Phe Gly Glu Ser Tyr Ile Val

545 550 555 560

Val Gly Ala Gly Glu Lys Ala Leu Lys Leu Ser Trp Phe Lys Lys Gly

565 570 575

Ser Ser Ile Gly Lys Met Phe Glu Ala Thr Ala Arg Gly Ala Arg Arg

580 585 590

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

595 600 605

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

610 615 620

Tyr Gly Val Leu Phe Ser Gly Val Ser Trp Thr Met Lys Ile Gly Ile

625 630 635 640

Gly Ile Leu Leu Thr Trp Leu Gly Leu Asn Ser Arg Ser Thr Ser Leu

645 650 655

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

660 665 670

Met Val Gln Ala

675

Claims

1.病毒样颗粒，其包含一种或更多种黄热病毒结构蛋白，其中所述黄热病毒结构蛋白的包膜区域含有氨基酸序列中的至少一个变化。

2.根据权利要求1所述的病毒样颗粒，其中所述至少一个变化在SEQ ID NO: 21的氨基酸位置182和氨基酸位置676之间，或在通过比对确定为对应于SEQ ID NO: 21的位置182和676的位置的位置之间。

3.根据权利要求1所述的病毒样颗粒，其中所述至少一个变化在由SEQ ID NO: 20表示的登革病毒1型的包膜区域的氨基酸位置90和氨基酸位置121之间，或在通过比对确定为对应于SEQ ID NO: 20的位置90和121的位置的位置之间。

4.根据权利要求3所述的病毒样颗粒，其中所述至少一个变化在SEQ ID NO: 20的氨基酸位置108，或在通过比对确定为对应于SEQ ID NO: 20的位置108的位置。

5.根据权利要求1至4中任一项所述的病毒样颗粒，其中所述黄热病毒为西尼罗河病毒、登革病毒、蜱媒脑炎病毒、黄热病病毒、日本脑炎病毒或丙型肝炎病毒。

6.根据权利要求5所述的病毒样颗粒，其中所述黄热病毒为登革病毒、西尼罗河病毒、日本脑炎病毒。

7.根据权利要求6所述的病毒样颗粒，其中所述黄热病毒为登革病毒。

8.根据权利要求1-7中任一项所述的病毒样颗粒，其中所述病毒结构蛋白包含prM和包膜区域。

9.根据权利要求1-7中任一项所述的病毒样颗粒，其中所述黄热病毒结构蛋白是包含由SEQ ID No.2、4、8、10、14和16中任一者表示的氨基酸序列的氨基酸位置16至氨基酸位置676的氨基酸序列的蛋白。

10.根据权利要求9所述的病毒样颗粒，其中所述黄热病毒结构蛋白是由SEQ ID No.2、4、8、10、14和16中任一者表示的氨基酸序列组成的蛋白。

11.根据权利要求1-10中任一项所述的病毒样颗粒，其中所述黄热病毒结构蛋白衍生自由与SEQ ID No.2、4、8、10、14和16中任一者表示的氨基酸序列具有90%或更高的序列同一性的氨基酸序列组成的蛋白。

12.分离的核酸分子，其包含核苷酸序列，所述核苷酸序列编码根据权利要求1-11中任一项所述的病毒样颗粒中含有的黄热病毒结构蛋白。

13.分离的核酸分子，其由与SEQ ID No.1、3、7、9、13和15中任一者表示的核苷酸序列具有90%或更高的序列同一性的核苷酸序列组成。

14.根据权利要求12所述的分离的核酸分子，其由SEQ ID No.1、3、7、9、13和15中任一者表示的核苷酸序列组成。

15.载体，其包含权利要求12所述的核酸分子，其中所述载体任选地包含可操作地连接至所述核酸分子的表达控制序列。

16.药物组合物，其包含：

(a)根据权利要求1-11中任一项所述的病毒样颗粒和/或根据权利要求12-15中任一项所述的核酸分子；以及

(b)药学上可接受的载体。

17.疫苗组合物，其包含根据权利要求1-11中任一项所述的病毒样颗粒。

18.DNA疫苗组合物，其包含根据权利要求12-15中任一项所述的核酸分子。

19.产生抗体的方法，其包括使根据权利要求1-11中任一项所述的病毒样颗粒和/或根据权利要求12-15中任一项所述的核酸分子与哺乳动物接触。

20.治疗或预防由黄热病毒感染引起的疾病或病况的方法，其包括将有效量的权利要求16-18中任一项所述的组合物施用于有需要的哺乳动物对象。

21.用于产生根据权利要求1-11中任一项所述的病毒样颗粒的方法，其包括培养用基因转染的细胞，所述基因编码根据权利要求1-11中任一项所述的病毒样颗粒中含有的至少一种黄热病毒结构蛋白；以及从细胞培养物回收病毒样颗粒。

22.权利要求1-11中任一项的病毒样颗粒，其用于诊断哺乳动物中的黄热病毒感染的方法或试剂盒中。