CN115667506A - 重组ace2-fc融合分子及其制备和使用方法 - Google Patents

重组ace2-fc融合分子及其制备和使用方法 Download PDF

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CN115667506A
CN115667506A CN202180007064.2A CN202180007064A CN115667506A CN 115667506 A CN115667506 A CN 115667506A CN 202180007064 A CN202180007064 A CN 202180007064A CN 115667506 A CN115667506 A CN 115667506A
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蔡宗義
麦雅丝
安德鲁·韦特
史蒂文·K·伦迪
马克·吉尔克里斯特
贾汉·哈利利
卓识
张勇
朱海
朱义
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Abstract

一种融合蛋白，其包含与Fc结构域共价融合的变体血管紧张素转化酶2(ACE2)结构域。变体ACE2结构域相对于具有SEQ ID NO.1的全长野生型ACE2包含N‑端缺失、C‑端缺失或两者。变体ACE2结构域具有ACE2活性。

Description

重组ACE2-FC融合分子及其制备和使用方法
相关申请的交叉引用
根据35 U.S.C.119(e)，本申请要求于于2020年2月13日提交的美国临时申请序列号62/976,344和于2020年10月1日提交的美国临时申请序列号63/086,593的申请日的优先权，其全部公开内容通过引用并入本文。
技术领域
本申请涉及预防或治疗涉及血管紧张素转换酶2(ACE2)的疾病、症状或病症，例如冠状病毒病2019(COVID-19)和相关病症。
背景技术
除非本文另有说明，否则本部分中描述的材料不是本申请中权利要求的现有技术，并且不因为包含在本部分中而承认是现有技术。
COVID-19是由严重急性呼吸综合征(severe acute respiratory syndrome，SARS)冠状病毒2(SARS-CoV-2)引起的传染病。COVID-19的并发症可包括长期肺损伤、肺炎、急性呼吸窘迫综合征(acute respiratory distress syndrome，ARDS)、外周和嗅觉神经损伤、多器官衰竭、败血性休克和死亡。2020年1月在《柳叶刀》上发表的对前41例确诊COVID-19病例的研究报告称，最早出现症状的日期为2019年12月1日。到2020年3月11日，世界卫生组织(WHO)宣布COVID-19爆发大流行病。截至2020年9月26日，188个国家和地区报告了超过3260万例病例，死亡人数超过99万，其中美国报告了超过750万例病例和20.5万例死亡病例。
除瑞德西韦(Remdesivir)外，尚未有药物或疫苗被批准用于治疗COVID-19的特定适应症。美国国立卫生研究院指南不建议在临床试验之外使用任何预防COVID-19的药物，无论是在接触SARS-CoV-2病毒之前还是之后。已由至少一个国家监管机构批准用于公共使用的九种疫苗：来自Pfizer-BioNTech和Moderna的两种RNA疫苗；来自Sinopharm、BharatBiotech和Sinovac的三种常规灭活疫苗；来自Sputnik V、Oxford-AstraZeneca和Janssen的三种病毒载体疫苗；以及一种肽疫苗(EpiVacCorona)。
血管紧张素转化酶2(ACE2)是一种含锌的金属酶，主要位于肺的肺泡细胞、小肠的肠细胞、动脉和静脉的内皮细胞、动脉的平滑肌细胞以及肺、动脉、心脏、肾、肠和其他组织中的其他细胞谱系的细胞膜上。ACE2通过平衡心血管、肾和呼吸系统中的血管紧张素转化酶活性来调节肾素-血管紧张素系统(renin angiotensin system，RAS)，表明了其在控制血压中的重要作用。ACE2在高血压、心脏功能、心功能和糖尿病的生理学中起保护作用。在急性呼吸窘迫综合征(ARDS)中，ACE、AngII和AT1R促进疾病发病机制，而ACE2和AT2R保护免于ARDS。此外，ACE2已被鉴定为严重急性呼吸综合征(SARS)冠状病毒的受体，并在严重急性呼吸综合征(SARS)发病机理中起关键作用。在冠状病毒家族中，至少有三种病毒SARS-CoV、MERS CoV和SARS-CoV-2使用它们的病毒蛋白之一(也称为Spike)结合人宿主细胞表面上的ACE2蛋白以用于病毒进入人体。
SARS-CoV-2是七种已知的感染人类的冠状病毒之一，包括SARS-CoV-1和MERS CoV病毒，其于2003年在亚洲和2012年在中东引起SARS爆发。对SARS-CoV-2病毒的免疫应答包括细胞介导的免疫和抗体产生的组合。尽管有超过1亿人从COVID-19中痊愈，但仍然不清楚在个体中对SARS-CoV-2病毒的自然免疫是否会持久。其中一个问题涉及病毒突变的持续累积，其可改变病毒抗原性谱并引起病毒突变株的再感染。截至2021年1月，在欧洲和南非鉴定的SARS-CoV-2病毒的变异株似乎传播如此迅速。这些变体菌株可能含有突变，这些突变最终增强病毒识别并感染到宿主细胞中。这些或其他可预见的变体是否可能降低疫苗的效力或克服天然免疫并导致再感染的发作仍然未知。
另一个关注点涉及抗体依赖性增强(ADE)的现象。当次优抗体(suboptimalantibody)的结合增强病毒进入宿主细胞时，就会产生ADE。在冠状病毒中，靶向病毒刺突(S)糖蛋白的抗体促进ADE。在SARS-CoV-1病毒的情况下，发现中和大多数变体的抗体能够增强突变病毒的免疫细胞进入，这反过来恶化了设计疫苗来抵抗的疾病。因此，ADE会妨碍疫苗开发，因为疫苗可能导致产生次优抗体。在这种情况下，在暴露于SARS-CoV-2病毒之前或之后，除了疫苗之外的任何预防策略都应被认为是规避ADE的可行替代方案。
因此，仍然非常需要有效治疗或预防涉及血管紧张素转化酶2(ACE2)的疾病或病症。
发明内容
以下概述仅是说明性的，并不旨在以任何方式进行限制。除了上述说明性方面、实施例和特征之外，通过参考附图和以下详细描述，其他方面、实施例和特征将变得显而易见。
本申请尤其提供了融合蛋白、融合蛋白复合物、蛋白质复合物、含有融合蛋白复合物的免疫缀合物和含有融合蛋白复合物的药物组合物。本申请还提供了制备融合蛋白和融合蛋白复合物的方法以及使用融合蛋白或融合蛋白复合物治疗或预防疾病的方法。
在一个方面，本申请提供了具有ACE2活性的融合蛋白。在一个实施方案中，融合蛋白包括与Fc结构域共价融合的变体血管紧张素转化酶2(ACE2)结构域。在一个实施方案中，变体ACE2结构域相对于全长野生型ACE2包含N-端缺失、C-端缺失或两者。在一个实施方案中，全长野生型ACE2结构域具有与SEQ ID NO.1具有至少95％、97％或98％的序列同一性的氨基酸序列。在一个实施方案中，变体ACE2结构域具有ACE2活性。
在一个实施方案中，变体ACE2结构域包含与来自全长野生型ACE2的氨基酸序列片段具有至少75％、80％、85％、90％、95％、97％、98％或99％的序列同一性的氨基酸序列。在一个实施方案中，该片段可以从选自全长野生型ACE2的残基1-17的氨基酸残基开始。在一个实施方案中，该片段可以以选自全长野生型ACE2的残基615-740的氨基酸残基结束。例如，变体ACE2结构域可以具有这样的氨基酸序列，其与来自全长野生型ACE2的残基1-615、残基2-618、残基2-740、残基4-615、残基17-615、残基17-740、或起始残基和终止残基的任何其他组合的氨基酸序列片段具有至少98％或99％的序列同一性。
在一个实施方案中，变体ACE2结构域包括与SEQ ID NO.3具有至少75％、80％、85％、90％、95％、97％、98％或99％的序列同一性的氨基酸序列。
在一个实施方案中，变体ACE2结构域可具有比全长野生型ACE2更高的对SARS-CoV或SARS Spike蛋白的结合亲和力。例如，变体ACE2结构域对SARS-CoV或SARS刺突蛋白(spike protein)可具有KD为0.1-100nM的结合亲和力(binding affinity)。
在一个实施方案中，变体ACE2结构域可具有比全长野生型ACE2更高的对SARS-CoV或SARS Spike蛋白的结合亲合力(binding avidity)。例如，变体ACE2结构域对SARS-CoV或SARS刺突蛋白可具有KD为0.01-10nM的结合亲合力。
在一个实施方案中，Fc结构域来源于自免疫球蛋白的Fc结构域。免疫球蛋白可以为IgG1、IgG2、IgG3、IgG4、IgA1(d-IgA1、S-IgA1)、IgA2、IgD、IgE或IgM。在一个实施方案中，Fc结构域可包含Fc铰链区。在一个实施方案中，Fc铰链区可被工程改造为C220S。在一个实施方案中，与野生型Fc结构域相比，Fc结构域可包含选自K322A、L234A和L235A的无效突变。在一个实施方案中，野生型Fc结构域具有具有SEQ ID NO.5具有至少98％或99％的序列同一性的氨基酸序列。
在一个实施方案中，Fc域可能缺乏效应子功能。在一个实施方案中，Fc结构域可能缺乏抗体依赖性细胞毒性(ADCC)、抗体依赖性细胞吞噬作用(ADCP)和补体依赖性细胞毒性(CDC)。在一个实施方案中，Fc结构域包含IgG1 Fc结构域。
在一个实施方案中，Fc结构域包含与SEQ ID NO.6具有至少75％、80％、85％、90％、95％、97％、98％或99％的序列同一性的氨基酸序列。
在一个实施方案中，融合蛋白可具有与SEQ ID NO.7、9、11、13、15、16、17、18、19和21具有至少75％、80％、85％、90％、95％、97％、98％或99％的序列同一性的氨基酸序列。
在一个实施方案中，融合蛋白的分子量可为约50-250kDa。在一个实施方案中，融合蛋白的分子量可为50kDa、60kDa、70kDa、80kDa、90kDa、100kDa、120kDa、150kDa、180kDa、200kDa、250kDa或其间的任何数值。
在第二方面，本申请提供了融合蛋白复合物。在一个实施方案中，融合蛋白复合物是本文公开的融合蛋白的同型二聚体。在一个实施方案中，融合蛋白复合物包含两个变体ACE2结构域。
在一个实施方案中，融合蛋白复合物包含至少两种融合蛋白。在一个实施方案中，两个融合蛋白通过一个或两个二硫键配对。在一个实施方案中，二硫键位于Fc结构域的铰链区。
在一个实施方案中，融合蛋白或融合蛋白复合物对SARS-CoV-2、SARS-CoV或SARS刺突蛋白或其片段具有结合亲和力。在一个实施方案中，结合亲和力的平衡解离常数不大于0.1nM、0.5nM、1nM、2nM、3nM、5nM、10nM、20nM、25nM、30nM、40nM、50nM、60nM、80nM或其间的任何数值。
在一个实施方案中，融合蛋白或融合蛋白复合物对SARS-CoV-2、SARS-CoV或SARS刺突蛋白或其片段具有结合亲合力。在一个实施方案中，结合亲合力的平衡解离常数不大于0.01nM、0.05nM、1nM、2nM、3nM、5nM、10nM或其间的任何数值。
在一个实施方案中，融合蛋白或融合蛋白复合物的比酶活性为约50-约5000pmol/min/μg。在一个实施方案中，融合蛋白的比酶活性为约568pmol/min/μg。
在第三方面，本申请提供了蛋白质复合物。在一个实施方案中，蛋白质复合物包括与病毒蛋白结合的融合蛋白或如其所公开的融合蛋白复合物。在一个实施方案中，病毒蛋白包含SARS-CoV-2、SARS-CoV、SARS刺突蛋白、冠状病毒、SARS病毒或其片段或组合。
在另一个方面，本申请提供了编码如其所公开的融合蛋白的分离的核酸。
在另一个方面，本申请提供了包含编码如其所公开的融合蛋白的分离的核酸的表达载体。
在另一个方面，本申请提供了包含编码如其所公开的融合蛋白的核酸的宿主细胞。在一个实施方案中，宿主细胞为原核细胞。在一个实施方案中，宿主细胞为真核细胞。
在另一个方面，本申请提供了用于产生如其所公开的融合蛋白和融合蛋白复合物的方法。在一个实施方案中，该方法包括用编码融合蛋白或融合蛋白复合物的核酸培养宿主细胞，从而产生融合蛋白或融合复合物。
在另一个方面，本申请提供了蛋白质缀合物(protein-conjugate)。在一个实施方案中，蛋白质缀合物包括融合蛋白或如其所公开的融合蛋白复合物和药物部分。药物部分可以通过接头与融合蛋白或融合蛋白复合物连接。在一个实施方案中，接头可以为选自酯键、醚键、胺键、酰胺键、二硫键、酰亚胺键、砜键、磷酸酯键、磷酯键、肽键、腙键或其组合的共价键。
在一个实施方案中，药物部分可以为抗病毒剂、免疫调节试剂、显像剂或其组合。在一个实施方案中，抗病毒剂可以是法匹拉韦(favipiravir)、利巴韦林(ribavirin)、加利地韦(galidesivir)、瑞德西韦(remdesvir)或其组合。在一个实施方案中，显像剂可以是放射性核素、荧光剂、量子点或其组合。
在另一方面，本申请提供了用于治疗涉及血管紧张素转化酶2(ACE2)的疾病或病症的药物组合物。在一个实施方案中，药物组合物物包括融合蛋白或如其所公开的融合复合物和药学上可接受的载体。在一个实施方案中，药物组合物还包括抗病毒剂。在一个实施方案中，药物组合物包含如其所公开的蛋白质缀合物和药学上可接受的载体。
在另一方面，本申请提供了治疗或预防受试者的病毒感染、急性呼吸窘迫综合征、肺动脉高血压或急性肺损伤的方法。在一个实施方案中，该方法包括向受试者施用有效量的本文公开的融合蛋白或融合复合物的步骤。在一个实施方案中，该方法还包括共施用有效量的治疗剂。在一个实施方案中，治疗剂包括抗病毒剂。在一个实施方案中，受试者是哺乳动物。
在一个实施方案中，病毒感染可以为SARS-CoV-2、SARS-CoV、SARS Spike蛋白、冠状病毒、SARS病毒或其片段或组合的感染。
在一个实施方案中，该方法可包括静脉内、皮下、通过鼻道(例如鼻喷雾剂)或通过肺通道施用融合蛋白或融合蛋白复合物。
在另一个方面，本申请提供了一种溶液。在一个实施方案中，该溶液包含有效浓度的本文公开的融合蛋白或融合蛋白复合物。在一个实施方案中，该溶液为受试者的血浆。在一个实施方案中，该溶液包含本文公开的融合蛋白、融合蛋白复合物或蛋白质复合物，并且该溶液为受试者的血浆。
附图说明
结合附图，根据以下描述和所附权利要求，本公开的前述和其他特征将变得更加显而易见。应当理解的是，这些附图仅描绘了根据本公开布置的若干实施例并且因此不应被认为是对其范围的限制，将通过使用附图以附加的特征和细节来描述本公开，其中：
图1显示了(1A)ACE2功能域和工程改造的Fc(无效)片段(SI-69R2和SI-69R4)之间的重组融合蛋白图，(1B)SI-F019融合蛋白的序列，缺乏N-端17-氨基酸信号肽的翻译后修饰的SI-69R2，(1C)指示SI-F019融合蛋白复合物是同型二聚体的尺寸排阻色谱，和(1D)SI-F019-Spike蛋白复合物图；
图2显示了SI-F019而非SI-69R4对TMPRSS2依赖性水解具有抗性(2A)，并且SI-F019的酶活性可在体外荧光测定中定量(2B)；
图3表明SI-F019对VeroE6细胞的活SARS-CoV-2感染的剂量依赖性阻断在测试的所有三种MOI的病毒中达到100％；
图4显示在0.01的MOI下，在SARS-CoV-2或SARS-CoV-1病毒感染1h后，以10fM或更高的浓度加入SI-F019保护了一部分Vero E6细胞免于经受细胞裂解；
图5显示了在与假病毒预孵育后，SI-F019以剂量依赖性方式抑制病毒感染并在较高浓度下实现完全抑制(IC50＝32.56nM)；
图6显示了内化/感染介导测定的结果，其中在THP1(pH7.2)(6A)、THP1(pH6.0)(6B)和Daudi(6C)中48h，当用测试浓度的SI-FO19预处理时，不存在指示假病毒(PsV)的GFP信号摄取，而低GFP信号与SI-69C1(抗S1抗体)和SI-69R3(SARS-CoV-2ACE-2Fc WT)以及培养基、缓冲液和ACE2-his(SI-69C1)相关；
图7显示了如通过指示PsV感染的GFP信号测量的，SI-F019可以与天然抗SARS-CoV-2抗体或ACE2-Fc(野生型)融合蛋白竞争以阻断Fc介导的抗体依赖性增强(ADE)；
图8显示了如通过使用抗Spike抗体和抗人Fc抗体检测的，表达SARS-CoV-2Spike蛋白的HEK293-T细胞的流式细胞仪分析；
图9显示了通过几何平均荧光强度(gMFI)测量的SI-F019与表达SARS-CoV-2Spike蛋白的HEK293-T细胞的剂量依赖性结合；
图10展示了抗体依赖性细胞毒性(ADCC)测定的FACS分析，显示如通过钙黄绿素-AM和碘化丙啶染色测量的，人抗S1抗体(SI-69C3)指导人NK细胞靶向表达SARS-CoV-2Spike蛋白的HEK293-T细胞；
图11显示当与人抗S1抗体(SI-69C3)相比时，SI-F019在100μM和100nM之间的治疗剂量下不介导ADCC，而其具有野生型Fc的变体(SI-69R3)以剂量依赖性方式介导ADCC，即使活性水平较低；
图12显示了如通过表达SARS-CoV-2S蛋白的HEK293-T细胞的活力所测量的，Fc无效突变使得SI-F019能够在体外降低血清介导的补体依赖性细胞毒性(CDC)；
图13显示了通过测量在各种处理后表达SARS-CoV-2S蛋白的HEK293-T细胞的活力，SI-F019在体外不诱导血清补体依赖性细胞毒性(CDC)(13A)；并且SI-F019的Fc无效突变在体外处理96h后对后续的细胞生长没有影响；以及
图14显示了SI-F019在PBMC培养物中不引起细胞因子以可溶性或平板结合形式的释放：(14A)IFNγ；(14B)TNFα；(14C)GM-CSF；(14D)IL-2；(14E)IL-10；(14F)IL-6；(14G)IL-1β；(14H)IL-12p70；以及(14I)MCP-1。
具体实施方式
在下面的详细描述中，参考了构成其一部分的附图。在附图中，除非上下文另有规定，否则相似的符号通常标识相似的组件。在详细描述、附图和权利要求中描述的说明性实施方案并不意味着是限制性的。在不脱离本文所呈现的主题的精神或范围的情况下，可利用其他实施方案，且可作出其他改变。将容易理解的是，如本文中一般性描述的以及在附图中示出的本公开的方面可以以各种各样的不同配置来布置、替换、组合、分离和设计，所有这些在本文都是明确地预期的。
本申请尤其涉及融合蛋白如重组人ACE2-Fc融合蛋白的产生和表征。在一些实施方案中，这些融合蛋白能够保护人宿主细胞的膜ACE2免受病毒颗粒或病毒的侵害。在一个实施方案中，病毒颗粒或病毒可以利用病毒刺突蛋白在感染后使病毒进入宿主细胞。在一个实施方案中，病毒颗粒包括但不限于SARS-CoV-2病毒、COVID-19病毒、SARS-CoV-2的变体和其他冠状病毒。在一个实施方案中，病毒可引起严重急性呼吸综合征(SARS)。在一个实施方案中，SARS可以包括冠状病毒病2019或COVID-19。
在一个实施方案中，重组人ACE2-Fc融合蛋白可以是ACE2锌金属肽酶结构域(也称为ACE2胞外结构域，ACE2-ECD)和IgG1Fc片段的融合蛋白。在一个实施方案中，融合蛋白是SI-F019、ACE2-ECD和IgG1 Fc片段的融合蛋白，具有根据EU编号系统的C220S、L234A、L235A和K322A的突变(表1和图1)。活性ACE2-ECD保持宿主受体-病毒相互作用的结构构象。IgG1Fc片段中的每个突变可削减某些免疫应答。突变C220S可去除未配对的半胱氨酸以配对重链和轻链，从而提供避免蛋白质形成聚集、提高蛋白质稳定性和提高生产效率和可扩展性等技术优势。L234A和L235A的引入可以降低Fc的效应子功能，例如抗体依赖性细胞毒性(ADCC)、抗体依赖性细胞吞噬作用(ADCP)。K322A突变可以降低C1q结合触发的补体依赖性细胞毒性(CDC)。SI-F019被设计用于中和SARS-CoV-2病毒，同时触发较少的效应子应答。
除非上下文不合适，否则本文所用的术语“一”、“一个”和“该”被定义为意指“一个或多个”并且包括复数。
术语“重组融合蛋白”是指通过对编码两个或多个最初编码不同蛋白质的基因的融合基因进行基因工程而产生的蛋白质。
术语“ACE2-Fc”指人ACE2蛋白片段和人免疫球蛋白的片段可结晶区(Fc区)的工程片段的重组融合蛋白，其中人免疫球蛋白包括但不限于IgG1、IgG2、IgG3、IgG4、IgA1(d-IgA1、S-IgA1)、IgA2、IgD、IgE和IgM。
术语“刺突蛋白(spike)”、“Spikes”、“S蛋白”或变体是指负责允许病毒附着(“S1亚基”或“S1蛋白”)并与宿主细胞膜融合(“S2亚基”或“S2蛋白”)的蛋白。在COVID-19的情况下，SARS-CoV-2对人细胞上的ACE2受体具有足够的亲和力以将它们用作细胞进入的机制，并且SARS-CoV-2对人ACE2的亲和力高于原始SARS病毒。
术语“Fc结构域”、“Fc片段”和“Fc区”是指来源于铰链的IgG、IgA和IgD抗体同种型中的Fc区的相同结构域或片段(分别为“Fc结构域”和“Fc片段”)，以及抗体的两条重链的第二和第三恒定结构域(CH2-CH3)。
术语“亲和力(affinity)”是指两种多肽之例如受体/配体、ACE2/刺突蛋白或其变体间吸引力的量度。两个多肽之间的内在吸引力可以表示为特定相互作用的结合亲和力平衡常数(KD)。KD结合亲和力常数可以例如通过生物膜层干涉技术来测量。
术语“亲合力(avidity)”是指单个非共价结合相互作用的多重亲和力的累积强度，例如在蛋白质受体与其配体之间，并且通常称为功能亲和力。因此，亲合力不同于描述单一相互作用的强度的亲和力。
术语“抗原性漂移(antigenic drift)”是指感染性病毒的随机遗传突变，其产生抗原性具有微小变化的新病毒株，防止由先前病毒株感染的抗体可能对该抗原性无效。
术语“细胞因子释放综合征”(CRS)是指在严重的COVID-19病例中与炎性介质水平升高相关的CRS，该炎性介质包括细胞因子和趋化因子，例如白介素(IL)-2、IL-6、IL-7、IL-10、肿瘤坏死因子(TNF)、粒细胞集落刺激因子(G-CSF)、单核细胞趋化蛋白-1(MCP1；也称为CCL2)、巨噬细胞炎性蛋白1α(MIP1α；也称为CCL3)、CXC-趋化因子配体10(CXCL10)、C-反应蛋白、铁蛋白和DSARS-CoV-2感染后的血液中-二聚体。
实施例
实施例1.重组ACE2-Fc融合蛋白的克隆、表达和纯化
人膜ACE-2是介导SARS-CoV病毒进入人宿主细胞的关键受体。人ACE2蛋白具有至少三个功能域：信号肽(残基1-17)、锌金属肽酶结构域(残基18-615)和TMPRSS2蛋白酶切割位点(残基697-716)(SEQ ID NO.1为来自Genbank号NP_001358344.1的全长人ACE2蛋白质序列)，其中SARS-CoV病毒蛋白、Spike与锌金属肽酶结构域相互作用(SEQ ID NO.3为残基1-615截短的ACE2的蛋白质序列。另一方面，人抗体的Fc区(SEQ ID NO.5)能够与许多免疫细胞上的Fc受体(FcR)和补体系统的一些蛋白质相互作用。IgG1 Fc区的每个Fc片段(根据EU编号系统)在C220含有半胱氨酸，其可以固有地与κ或λ轻链形成二硫键。为了降低具有可能使蛋白质去稳定和/或失活的游离半胱氨酸的风险，C220可由丝氨酸(C220S)或其他氨基酸取代。为了减少Fc与FcγR和C1q的结合，可以将其他点突变如K322A、L234A和L235A工程改造成野生型IgG1 Fc片段。携带四个突变的IgG1 Fc片段统称为IgG1 Fc无效(SEQ IDNO.6)。
重组人ACE2-Fc融合蛋白(如表1所列)经工程改造以产生可溶性融合蛋白，其中SI-69R2(SEQ ID NO.7)为不具有TMPRSS2蛋白酶切割位点的截短的ACE2片段与IgG1Fc无效片段的重组融合蛋白。产生其他重组融合蛋白以提供Ig同种型的Fc片段，如SI-69R2-G4(IgG4 Fc，SEQ ID NO.9)、SI-69R2-A1(IgA1 Fc,SEQ ID NO.11)、SI-69R2-A2(IgA2 Fc，SEQID NO.13)、或野生型IgG1 Fc片段(IgG1 Fc，SEQ ID NO.19)。还产生了具有所有三个结构域和野生型IgG1 Fc片段的截短的ACE2的重组融合蛋白(SI-69R4，1-740，SEQ ID NO.21)。在所有重组ACE2-Fc融合蛋白中，信号肽(ACE2残基1-17)可以用不同长度的其他信号肽替换，而不影响人ACE蛋白或ACE2-Fc融合蛋白中其他结构域的功能。
将表1中编码融合蛋白的重组融合基因克隆到pCGS3.0(如SI-69R2)或pTT5表达载体(如SI-69R4和SI-69R10)中并在ExpiCHO细胞中表达。按照标准蛋白表达方案纯化所有融合蛋白，使用0.22μm滤器灭菌，并保存在4℃的冷冻保存缓冲液中。在表达和纯化过程中，每个重组融合蛋白可以进行翻译后修饰，包括N-糖基化和N-端信号肽(17个氨基酸)的裂解。在SI-69R2的情况下，纯化的融合蛋白被赋予新的名称SI-F019。
如图1A和1B所示，SI-F019保留了截短的ACE2片段(残基18-615)，其包含人ACE2的锌金属肽酶结构域(残基19-611)，但不包含TMPRSS2蛋白酶切割位点。此外，SI-F019保留了没有与Fcγ受体结合的IgG1 Fc无效片段。这样，预期可溶性形式的SI-F019不会结合外周血中的任何靶细胞。
SI-F019融合蛋白可能进行翻译后修饰，例如N-糖基化和通过Fc区的两个二硫键连接的同源二聚化。为了评估SI-F019二聚体的实际分子量，在多角度光散射(MALS)、吸光度(UV)和/或折射率(RI)浓度检测器技术的组合中使用分析型尺寸排阻色谱法(SEC)，如图1C中所示。该方法结合了通过分子大小的色谱分离和通过光散射(LS)的绝对摩尔质量的测定，而不受分子量标准校准的限制。SI-F019表现出209.6kDa的平均总分子量(主峰)，其中SI-F019二聚体及其修饰物(即聚糖)的分子量分别测量为189.3kDa和20.3kDa。在其氨基酸的理论计算中，SI-F019单体的分子量为95.1kDa。因此，纯化的SI-F019融合蛋白复合物是同型二聚体，而SI-F019蛋白复合物是指作为单体或二聚体的SI-F019与其他蛋白如刺突蛋白和效应蛋白之间的蛋白-蛋白相互作用。SI-F019-Spike蛋白复合物的形成(如图1D所示)是该机制的基础，通过该机制，SI-F019作为一种候选抑制剂可防止SARS-CoV-2病毒与膜ACE2对接以使病毒进入宿主人体细胞。
实施例2：SI-F019对刺突、Fc受体和C1q的结合亲和力
SI-F019被设计成通过阻止刺突蛋白与人宿主细胞上的膜ACE2蛋白结合来阻断SARS-CoV病毒进入人体。刺突是冠状病毒最显著的特征，其是负责冠状或环状表面的球状结构。刺突蛋白通常由糖蛋白组成，每个刺突由S蛋白的三聚体组成，而S蛋白又由S1和S2亚基组成。同源三聚体S蛋白介导病毒和宿主细胞之间的受体结合和膜融合。S1亚基形成刺突的头部并具有受体结合结构域(RBD)。S2亚基形成茎，其将刺突锚定在病毒包膜中，并且在蛋白酶激活时能够融合。在功能活性状态下，当病毒在宿主细胞的蛋白酶如组织蛋白酶家族和跨膜蛋白酶丝氨酸2(TMPRSS2)的作用下与宿主细胞结合并融合时，S1和S2的亚基复合物分裂成单个亚基。刺突在冠状病毒感染过程的病毒进入中起重要作用。在COVID-19的情况下，SARS-CoV-2病毒与宿主细胞表面上的膜结合的ACE2受体对接，刺突和ACE2的功能域之间的相互作用通过触发病毒包膜和宿主细胞膜之间的融合而引起病毒核衣壳释放到宿主细胞细胞质中。
评估了SI-F019的ACE-Fc融合蛋白与病毒刺突蛋白的结合亲和力和亲合力。在生物膜层干涉技术分析中，刺突蛋白的样品包括SARS-CoV-2刺突三聚体、SARS-CoV-2S1蛋白、SARS-CoV-2S1蛋白RBD结构域和SARS-CoV-1RBD结构域(表2)。结合亲和力测定测量了固定在抗人IgG Fc捕获生物传感器尖端表面上的SI-F019与溶液中的刺突蛋白的结合。亲合力测定测量了固定在链霉抗生物素生物传感器尖端表面上的生物素化刺突蛋白与溶液中的SI-FO19的结合。数据分析使用1:1拟合模型计算结合亲和力和亲合力。结果表明SI-F019对这些刺突蛋白、片段或结构域的结合亲和力和亲合力似乎在它们各自的以纳摩尔(nM)计的KD范围内(表2)。该特征和信息数据可以是用于测量具有指示SARS-CoV-2变体中潜在抗原漂移的病毒刺突蛋白变体的SI-F019蛋白复合物的有用参考。该现象与实际情况接近，涉及SARS-CoV-2病毒的某些毒株中的病毒突变，例如刺突蛋白中的D614G(Zhang等人，2020)，其可能改变了病毒对膜ACE2的亲和力和病毒进入宿主细胞。
与其结合至刺突并行，通过使用生物膜层干涉技术评估SI-F019与人FcγR、C1q和FcRn的结合。如表3所示，未检测到与FcγR(包括FcγRI、FcγRIIa、FcγRIIb和FcγRIIIa)的结合，也未检测到与C1q的结合。然而，SI-F019确实与FcRn结合，在KD为37.6nM时测定结合亲和力，其与人IgG1 Fc区相当。
实施例3.SI-F019对TMPRSS2蛋白酶活性具有抗性
人ACE2通过TMPRSS2进行膜蛋白酶水解，单体细胞外ACE2从细胞中脱落，这可以在血清中容易检测到。在重组ACE2-Fc融合蛋白中，截短的ACE2结构域与Fc片段融合，但仍保持与病毒刺突蛋白的结合亲和力。
SI-F019被工程改造为在截短的ACE2结构域中不含TMPRSS2切割位点。如图1所示，SI-F019含有残基18-615，而SI-69R4编码包括TMPRSS2切割位点的所有三个ACE2结构域(残基：1-740，SEQ ID NO.21)。为了证明SI-F019没有TMPRSS2特异性蛋白水解，将SI-69R4用作对照。为了进行TMPRSS2特异性水解的测定，根据Genbank：NP_001358344.1克隆、表达并纯化TMPRSS2(106-492)催化结构域。如图2A所示，在不存在TMPRSS2的情况下，SI-F019和SI-69R4均稳定地迁移至它们各自的分子量(作为变性条件下的单体)。当添加TMPRSS2时，SI-F019显示出其对TMPRSS2的抗性，而SI-69R4进行了蛋白水解，表明其对TMPRSS2的敏感性与预期的一致。因此，SI-F019是稳定的并且对TMPRSS3介导的蛋白酶活性具有抗性。
实施例4.SI-F019发挥ACE2的酶活性
SI-F019是截短的ACE2(残基18-615)和IgG1 Fc无效片段的融合蛋白。截短的ACE2编码一种锌金属肽酶，其酶活性可以通过使用已建立的测定法重新评估。具有MCA(7-甲氧基香豆素-4-乙酸)荧光标签的ACE2的肽底物[MCA-YVADAPK(Dnp)-OH荧光肽底物]用于测量SI-F019的ACE2酶活性。制备MCA分子作为游离荧光团定量的标准曲线校准，并将底物在DMSO中稀释至0.97mg/ml。将SI-F019稀释至100、200和300ng/ml并用于体外裂解荧光肽以释放游离MCA。在室温下将该测定孵育20分钟，并在2分钟间隔的时间点收集荧光信号的数据。
使用MCA标准曲线以摩尔计对裂解的MCA进行定量。根据如图2B所示的线性曲线的斜率(MCA量对时间)测定酶活性。SI-F019在所有三种浓度下显示良好的线性(R2>0.99)，表明肽的稳定裂解是浓度依赖性的。为了计算酶活性，将斜率除以SI-F019的质量数(μg)。最终的比酶活性为568pmol/min/g。SI-F019保留了膜ACE2的酶活性的事实表明ACE2的独立结构域也保留了宿主受体-病毒相互作用的结构构象。
实施例5.SI-F019抑制活SARS-CoV-2感染VeroE6细胞。
测试SI-F019体外抑制活SARS-CoV-2感染和VeroE6(ATCC：CRL-1586)细胞裂解的能力。将1.5-1200nM的SI-F019测试浓度与3种浓度的活SARS-CoV-2病毒(USA-WA1/2020株，代表100倍范围的感染复数，MOI)预孵育1h，然后加入到90％汇合的VeroE6细胞单层中。1h后，去除含有病毒的培养基，并替换为含有匹配测试浓度的SI-F019的培养基，测试一式三份进行。72h后通过中性红染料的摄取来测量细胞活力，并通过与其中在没有SI-F019的情况下以各MOI加入病毒的孔比较确定裂解性病毒感染的抑制百分比。使用GraphPad Prism软件计算各病毒浓度(1MOI＝40,000个病毒颗粒)的50％抑制浓度(IC50)，并显示在每张图上。SI-F019与活SARS-CoV-2的预孵育导致感染的剂量依赖性阻断，在测试的所有三种MOI的病毒中达到100％。如图3所示，SI-F019在1.0的MOI下中和多达40,000个病毒颗粒，IC50为97.62nM。在0.1和0.01的MOI下，SI-F019能够分别在79.95nM和36.5nM的IC50下阻断感染。
实施例6.SI-F019减少病毒复制和再感染
测试了SI-F019抑制复制和再感染的能力，即从以前感染了低MOI的SARS-CoV-2或SARS-CoV-1病毒的细胞向VeroE6细胞进一步转移感染的能力。将90％汇合单层中的VeroE6细胞(约20,000个细胞)暴露于SARS-CoV-2(USA-WA1/2020株)或SARS-CoV-1(Urbani2003000592株)1h，MOI为0.01(计算为400个病毒感染性颗粒)。洗去游离病毒颗粒后，将SI-F019以10fM-100nM的范围一式三份加入细胞中，并将细胞培养物保持72h。通过中性红染料摄取来测定细胞活力，并计算病毒细胞毒性的抑制％。使用无病毒或无药物条件(NVND)下的最大吸光度表示100％细胞活力，在每个平板上归一化吸光度值，并使用下式确定病毒/无药物条件(VND)下的平均吸光度值：
细胞存活％＝[(孔OD540-VND OD540)/(NVND OD540-VND OD540)]*100
如图4所示，以10fM的浓度加入SI-F019保护Vero E6细胞免受二次感染。以0.01的MOI感染SARS-CoV-2或SARS-CoV-1病毒1h的培养物使细胞裂解减少至少20％。然而，当SI-F019的浓度以10倍增量增加至100nM时，在该测定中未观察到保护的显著增加。该发现表明，向感染低滴度病毒的细胞中加入SI-F019可以降低病毒的传播以及细胞毒性的程度，即使在低浓度下也是如此。
实施例7.SI-F019抑制HEK293T-ACE2细胞的假病毒感染
HEK293T(ATCC:CRL-3216)-3D4通过慢病毒转导人ACE2蛋白产生克隆细胞系。表达的人ACE2的功能通过酶底物转化测定并通过FACS的特异性抗体的结合来证实。含有荧光素酶报道基因的SARS-CoV-2S蛋白包装的假病毒获自国家药品生物制品检定所(NationalInstitute for the Control of Pharmaceutical&Biological Products)。根据制造商的说明书进行测试。将S-假病毒原液在MRD为20的培养基中稀释，以产生300TCID50/孔的病毒载量。将浓度为0.07-1500nM的SI-F019与稀释的病毒溶液预孵育1h。将HEK293T-3D4细胞分散在96孔板中。1h后，将混合物加入到细胞板中。孵育24h后，通过测试荧光素酶活性来测量感染的细胞。使用GraphPad Prism软件计算确定病毒载量的50％抑制浓度(IC50)。图5显示了在与假病毒预孵育后，SI-F019以剂量依赖性方式抑制病毒感染并在较高浓度下实现完全抑制(IC50＝32.56nM)。
实施例8.SI-F019降低ADE的发生率
抗体依赖性增强(ADE)是病毒与次优抗体的结合增强其进入宿主细胞的现象。在COVID-19的情况下，SARS-CoV-2病毒对具有由初次感染产生的抗SARS-CoV-2抗体的患者或对已接种疫苗的个体的继发感染可导致单核细胞和B细胞对病毒的摄取增加。与病毒接触的抗病毒抗体可以结合在某些免疫细胞或某些补体蛋白上表达的Fc受体。后者的结合依赖于抗体的Fc区。通常，病毒在称为吞噬作用的过程中降解，由此病毒颗粒被宿主细胞通过质膜吞噬。然而，如果病毒没有被抗体中和，由于低亲和力结合或靶向非中和表位，抗体结合可能导致病毒逃逸。结果是抗体增强的感染。
通过天然免疫或接种产生的抗体具有野生型Fc区。虽然SI-F019能够与抗刺突抗体竞争结合SARS-CoV2病毒，但是IgG1 Fc无效片段不能结合Fc受体或C1q(参见表3)。为了证明SI-F019在降低ADE作用方面的比较优势，评估了SI-F019在内化、复制和再感染中的作用。
在测量Fc介导的内化的试验中，将SARS-CoV-2S蛋白包装到表达GFP的假病毒(PsV)中，表达Fc受体和补体受体2(CR2)的两种细胞系THP1(单核细胞)和Daudi(B细胞)用于测试FcRγ和CR2介导的ADE机制。SI-69R3用作SI-F019的对照，其具有野生型Fc，与具有IgG1 Fc无效修饰的SI-F019相反(参见表1)。在暴露于PsV 48h后，将来自细胞的绿色荧光信号定量为PsV感染的指示。在用PsV和SI-69C1处理的条件下，在THP1(pH7.2)(6A)、THP1(pH6.0)(6B)和Daudi(6C)细胞中测量48h的抗S1抗体或SI-69R3低水平的绿色荧光。该结果表明PsV的一些转移可通过Fc受体发生。相反，SI-F019在所示浓度的条件下导致THP1或Daudi细胞没有摄取PsV，与在阴性对照条件(包括测定培养基、制剂缓冲液和SI-69C1)中测量的绿色荧光信号相当(图6)。Fc介导的ADE的作用是剂量依赖性的，因为对细胞的处理使用1pM-100nM的剂量进行。这表明通过FcγR或CR2机制可能摄取一些PsV。
实施例9.SI-F019降低PsV的病毒载量
由于缺乏功能性Fc片段，SI-F019可能不介导S蛋白包装的表达GFP的假病毒(PsV)的内化。为了确定SI-F019是否可以抑制假病毒的摄取，SI-F019以竞争模式与SI-69R3或天然抗SARS-CoV-2抗体共同处理。在感染相同组的靶细胞之前，将PsV与SI-F019以1pM-100nM的剂量范围与10pM的抗SARS-CoV-2(S1)抗体或10pM的SI-69R3一起孵育1h。检测PsV来源的GFP信号作为感染的病毒载量。SI-F019能够从10fM开始抑制靶细胞中PsV的病毒载量(图7)。
虽然诸如抗SARS-CoV-2(S1)抗体的抗体和SI-69R3中的截短的ACE2-野生型Fc片段的融合蛋白都显示能够介导SARS-CoV-2刺突假型慢病毒的内化，但是SI-F019由于缺乏功能性Fc片段而无法介导内化。本文中，SI-F019在10pM抗SARS-CoV-2(S1)抗体或10pM SI-69R3的存在下，甚至在10fM的低浓度下也有助于降低PsV的病毒载量。总之，这些结果表明SI-F019可以分别降低THP1单核细胞和DaudiB细胞中由FcRγ和CR2依赖性机制诱导的ADE的发生率。
实施例10.表达SARS-CoV-2刺突蛋白的HEK293-T细胞
通过转导用编码SARS-CoV-2刺突蛋白的cDNA(登录号：YP_009724390.1)包装的慢病毒并表达IRES，并基于由相同表达构建体(LPP-CoV219-Lv105-050,GeneCopoeia)驱动的嘌呤霉素抗性进行选择，建立稳定表达SARS-CoV-2刺突蛋白的HEK293-T细胞(ATCC：CRL-3216)。SARS-CoV-2刺突蛋白的表达通过结合人IgG克隆AM001414来证实，其对SARS-CoV-2Spike蛋白“抗Spike”(SKU938701，Biolegend)具有特异性，并且人IgG同种型匹配的克隆QA16A12用作对照“同种型”(SKU403502，Biolegend)。如图8所示，通过与多克隆抗人FcAF647 Fab(SKU109-607-008，Jackson ImmunoResearch)二次孵育和FACS评估来定量结合的蛋白质。
将表达SARS-CoV-2刺突蛋白的HEK293-T细胞和亲本HEK293细胞于37℃在内化抑制剂叠氮化钠存在下用指定材料染色30分钟。去除游离的SI-F019后，使用抗人Fc AF647fab(SKU109-607-008，Jackson ImmunoResearch)和流式细胞仪分析来检测和定量SI-F109。几何平均信号强度用于定量SI-F019与靶细胞系的结合，如图9所示。表达SARS-CoV-2刺突蛋白的HEK293-T细胞可用作COVID-19感染细胞的模型。
实施例11.SI-F019对抗体依赖性细胞毒性(ADCC)的影响
抗体依赖性细胞毒性(ADCC)是对病毒感染的重要免疫应答之一，例如在COVID-19的情况下感染SARS-CoV-2病毒。在最初的病毒感染后，抗病毒抗体直接结合病毒颗粒以用于中和和凝集。病毒-抗体复合物与吞噬细胞上的Fc受体的结合可触发吞噬作用，导致病毒的破坏；与免疫效应细胞(例如单核细胞、嗜中性粒细胞、嗜酸性粒细胞和NK细胞)上的Fc受体结合可触发细胞毒性因子(例如抗病毒干扰素)的释放，产生对病毒复制有害的微环境。
为了区分SI-F019与抗刺突抗体的作用，用钙黄绿素-AM装载表达SARS-CoV-2刺突蛋白的HEK293-T细胞，并与纯化的人NK细胞以5:1的效应子与靶标比率共培养。测试的处理包括SI-F019和S1特异性人IgG克隆SI-69C3。SI-69C3是人抗体克隆CC12.3，分离自住院的COVID-19患者(10.1126/science.abc7520)。共培养12h后，将细胞用碘化丙啶染色并评估活力。如图10所示，基于钙黄绿素-AM和碘化丙啶染色的表达的活靶细胞频率(群体3)的降低被评估为细胞溶解的量度。
当暴露于S1特异性人IgG克隆SI-69C3(克隆CC12.3)时，NK细胞介导的ADCC可针对表达SARS-CoV-2蛋白的HEK293-T细胞。在100nM-100fM的处理范围内，与SI-69C3相比，SI-F019不介导ADCC。在这些测定条件下，具有野生型Fc的SI-F019药物变体(SI-69R3)能够以剂量依赖性方式介导ADCC，但活性水平与S1特异性人IgG克隆CC12.3相比更低，如图11和图12所示。这些数据表明，与SARS-CoV-2S1特异性人IgG抗体不同，SI-F019不介导NK细胞介导的ADCC。
实施例12.SI-F019对补体依赖性细胞毒性(CDC)的影响
补体级联在介导COVID-19患者中基于抗体的细胞和组织损伤中的作用在天然免疫应答和基于中和抗体的治疗中都是明显的(Perico等人，2021)。由病毒和特异性IgG形成的免疫复合物介导补体诱导的血液凝固、血栓栓塞和全身性微血管病。COVID-19患者中的这些广泛并发症可能危及生命，并且依赖于与IgG结合的补体蛋白。通过C1q桥接红细胞的病毒免疫复合物和具有FcγRIIA的血小板是COVID-19患者中血栓栓塞的介质(Nazy等人，2020)。免疫复合物对内皮血管壁的固定和补体介导的凝血是COVID-19患者的主要关注点，其中内皮细胞的活化是血栓栓塞级联的一部分。
与天然IgG抗体不同，SI-F019不能结合C1q，如表3所示。该特征消除了感染的上皮和内皮诱导细胞死亡的风险，这些上皮和内皮可能在其表面瞬时表达SARS-CoV-2刺突蛋白。SI-F019的这种保护作用与抗刺突人IgG抗体相比得到证实。
为了证明SI-F019的保护作用，将表达SARS-CoV-2刺突蛋白的HEK293-T细胞在无血清培养基(Optimem)中培养处理30分钟，然后以1:10的血清与培养基比率加入人血清补体。测试的处理包括SI-F019和S1特异性人IgG克隆AM001414。将细胞在37℃下培养3h，然后加入碘化丙啶染色并在每个孔中计数阳性染色细胞。如图12和图13所示，用Incucyte Zoom软件在3h计数的红细胞作为CDC的量度进行评估。如图13所示，96h后评估总细胞汇合，作为CDC影响的量度。
通过这些细胞在人血清补体攻击后进一步增殖的能力进一步证实了组织细胞免受补体损伤的保护。当暴露于S1特异性人IgG克隆(克隆AM4141)时，由与无血清培养基的1:10体积比的人血清补体介导的CDC可针对表达SARS-cov-2S蛋白的HEK293T细胞来评估。结果表明人可溶性单体ACE2和SI-F019都不介导CDC，而SI-69R3与人IgG抗体相比具有有限的剂量依赖性的CDC活性增加。基于处理后96h的孔汇合，CDC细胞溶解反映为细胞生长减少。
实施例13.由PBMC培养物中的可溶性或板结合的SI-F019引起的细胞因子释放。
SARS-CoV-2具有针对呼吸道和小肠的ACE2表达上皮的趋向性。升高的IL-2、IL-6、IL-7、粒细胞-巨噬细胞集落刺激因子(GM-CSF)、干扰素γ诱导蛋白10(IP-10)、单核细胞趋化蛋白1(MCP-1)、巨噬细胞炎性蛋白1-α(MIP-1α)和指示细胞因子释放综合征(CRS)的肿瘤坏死因子-α(TNF-α)的临床实验室发现表明了潜在的免疫病理学。CRS是主要的不良副作用，其可以限制使用生物制剂的处理的效用，并且使用体外细胞因子释放测定进行测试。
SI-F019是由人ACE2和不能结合Fcγ受体的人IgG1 Fc的突变形式组成的融合蛋白。因此，预期SI-F019不会结合外周血中的任何靶细胞或引起细胞因子释放。将从5个健康供体分离的包括嗜中性粒细胞的白细胞(WBC)置于含有2000nM和200nM浓度的板结合或可溶性SI-F019的培养孔中。
作为阳性对照，TGN1412抗体以相同的浓度和相同的形式使用，这是由于其在该测定的板结合形式中诱导细胞因子释放的充分证明的能力。通过与具有能够结合由外周血中几种细胞类型表达的Fcγ受体的野生型Fc片段的SI-69R3比较，评估IgG1 Fc无效片段减少细胞因子释放的潜在贡献。仅含有相似稀释度的SI-F019制剂缓冲液的WBC培养物用作阴性对照。在24h和48h时间点收集培养物上清液，并使用Meso Scale Discovery(MSD)平台检测9种细胞因子的存在。
如图14A-14E所示，细胞因子组中包括T细胞相关细胞因子IFNγ、TNFα、GM-CSF、IL-2和IL-10。如图14F-14I所示，还测试了促炎性非T细胞相关细胞因子IL-1β、IL-12p70和IL-6以及单核细胞趋化蛋白MCP-1的水平。取每个血液供体的重复孔的结果的平均值，并使用JMP14软件绘制成框图，显示95％置信区间和离群值。
结果表明，在200nM和2000nM浓度下，SI-F019不诱导任何测试的细胞因子暴露于板结合或可溶性形式的WBC。在所有条件下，SI-F019处理的样品中的细胞因子水平显示与缓冲液对照相似的浓度。阳性对照TGN1412强烈诱导板结合形式中的大多数细胞因子，但不诱导可溶性形式，这与先前公布的结果一致。当使用板结合的ACE2-Fc野生型刺激WBC时，可检测到IFNγ、GM-CSF和TNFα的一些中间产物，表明SI-F019的Fc无效片段的安全性增加。
最近在接受介入性IgG治疗的患者中提出了针对SARS-CoV-2病毒的体液反应的致病作用(Weinreich等人，2021；Chen等人，2021)。小血管高炎症反应是不良事件的基础，包括血小板增多症、瘙痒、发热和高血压。本申请表明，SI-F019可提供与IgG治疗相当的病毒中和的益处，同时保护组织和器官免于多种途径的功能障碍。因此，SI-F019可用于治疗、预防或缓和病毒感染，具体地用于预防和控制COVID-19的进展，同时减少临床并发症，并且另外用于急性呼吸窘迫综合征、肺动脉高血压或急性肺损伤。
表格
表1重组ACE2-Fc融合蛋白的克隆、表达和纯化
Figure GDA0003945357980000151
表2 SI-F019结合病毒蛋白的亲和力和亲合力
Figure GDA0003945357980000152
表3 Fc无效突变对其与Fc受体结合的影响
Figure GDA0003945357980000153
序列表
Figure GDA0003945357980000161
>序列ID 1：huACE2全长蛋白质序列(Genbank_number：NP_001358344.1，TMPRSS2蛋白酶切割位点)MSSSSWLLLSLVAVTAAQSTIEEQAKTFLDKFNHEAEDLFYQSSLASWNYNTNITEENVQNMNNAGDKWSAFLKEQSTLAQMYPLQEIQNLTVKLQLQALQQNGSSVLSEDKSKRLNTILNTMSTIYSTGKVCNPDNPQECLLLEPGLNEIMANSLDYNERLWAWESWRSEVGKQLRPLYEEYVVLKNEMARANHYEDYGDYWRGDYEVNGVDGYDYSRGQLIEDVEHTFEEIKPLYEHLHAYVRAKLMNAYPSYISPIGCLPAHLLGDMWGRFWTNLYSLTVPFGQKPNIDVTDAMVDQAWDAQRIFKEAEKFFVSVGLPNMTQGFWENSMLTDPGNVQKAVCHPTAWDLGKGDFRILMCTKVTMDDFLTAHHEMGHIQYDMAYAAQPFLLRNGANEGFHEAVGEIMSLSAATPKHLKSIGLLSPDFQEDNETEINFLLKQALTIVGTLPFTYMLEKWRWMVFKGEIPKDQWMKKWWEMKREIVGVVEPVPHDETYCDPASLFHVSNDYSFIRYYTRTLYQFQFQEALCQAAKHEGPLHKCDISNSTEAGQKLFNMLRLGKSEPWTLALENVVGAKNMNVRPLLNYFEPLFTWLKDQNKNSFVGWSTDWSPYADQSIKVRISLKSALGDKAYEWNDNEMYLFRSSVAYAMRQYFLKVKNQMILFGEEDVRVANLKPRISFNFFVTAPKNVSDIIPRTEVEKAIRMSRSRINDAFRLNDNSLEFLGIQPTLGPPNQPPVSIWLIVFGVVMGVIVVGIVILIFTGIRDRKKKNKARSGENPYASIDISKGENNPGFQNTDDVQTSF
>序列ID 2：huACE2全长DNA序列
(Genbank_number：NM_021804.3)
ATGTCAAGCTCTTCCTGGCTCCTTCTCAGCCTTGTTGCTGTAACTGCTGCTCAGTCCACCATTGAGGAACAGGCCAAGACATTTTTGGACAAGTTTAACCACGAAGCCGAAGACCTGTTCTATCAAAGTTCACTTGCTTCTTGGAATTATAACACCAATATTACTGAAGAGAATGTCCAAAACATGAATAATGCTGGGGACAAATGGTCTGCCTTTTTAAAGGAACAGTCCACACTTGCCCAAATGTATCCACTACAAGAAATTCAGAATCTCACAGTCAAGCTTCAGCTGCAGGCTCTTCAGCAAAATGGGTCTTCAGTGCTCTCAGAAGACAAGAGCAAACGGTTGAACACAATTCTAAATACAATGAGCACCATCTACAGTACTGGAAAAGTTTGTAACCCAGATAATCCACAAGAATGCTTATTACTTGAACCAGGTTTGAATGAAATAATGGCAAACAGTTTAGACTACAATGAGAGGCTCTGGGCTTGGGAAAGCTGGAGATCTGAGGTCGGCAAGCAGCTGAGGCCATTATATGAAGAGTATGTGGTCTTGAAAAATGAGATGGCAAGAGCAAATCATTATGAGGACTATGGGGATTATTGGAGAGGAGACTATGAAGTAAATGGGGTAGATGGCTATGACTACAGCCGCGGCCAGTTGATTGAAGATGTGGAACATACCTTTGAAGAGATTAAACCATTATATGAACATCTTCATGCCTATGTGAGGGCAAAGTTGATGAATGCCTATCCTTCCTATATCAGTCCAATTGGATGCCTCCCTGCTCATTTGCTTGGTGATATGTGGGGTAGATTTTGGACAAATCTGTACTCTTTGACAGTTCCCTTTGGACAGAAACCAAACATAGATGTTACTGATGCAATGGTGGACCAGGCCTGGGATGCACAGAGAATATTCAAGGAGGCCGAGAAGTTCTTTGTATCTGTTGGTCTTCCTAATATGACTCAAGGATTCTGGGAAAATTCCATGCTAACGGACCCAGGAAATGTTCAGAAAGCAGTCTGCCATCCCACAGCTTGGGACCTGGGGAAGGGCGACTTCAGGATCCTTATGTGCACAAAGGTGACAATGGACGACTTCCTGACAGCTCATCATGAGATGGGGCATATCCAGTATGATATGGCATATGCTGCACAACCTTTTCTGCTAAGAAATGGAGCTAATGAAGGATTCCATGAAGCTGTTGGGGAAATCATGTCACTTTCTGCAGCCACACCTAAGCATTTAAAATCCATTGGTCTTCTGTCACCCGATTTTCAAGAAGACAATGAAACAGAAATAAACTTCCTGCTCAAACAAGCACTCACGATTGTTGGGACTCTGCCATTTACTTACATGTTAGAGAAGTGGAGGTGGATGGTCTTTAAAGGGGAAATTCCCAAAGACCAGTGGATGAAAAAGTGGTGGGAGATGAAGCGAGAGATAGTTGGGGTGGTGGAACCTGTGCCCCATGATGAAACATACTGTGACCCCGCATCTCTGTTCCATGTTTCTAATGATTACTCATTCATTCGATATTACACAAGGACCCTTTACCAATTCCAGTTTCAAGAAGCACTTTGTCAAGCAGCTAAACATGAAGGCCCTCTGCACAAATGTGACATCTCAAACTCTACAGAAGCTGGACAGAAACTGTTCAATATGCTGAGGCTTGGAAAATCAGAACCCTGGACCCTAGCATTGGAAAATGTTGTAGGAGCAAAGAACATGAATGTAAGGCCACTGCTCAACTACTTTGAGCCCTTATTTACCTGGCTGAAAGACCAGAACAAGAATTCTTTTGTGGGATGGAGTACCGACTGGAGTCCATATGCAGACCAAAGCATCAAAGTGAGGATAAGCCTAAAATCAGCTCTTGGAGATAAAGCATATGAATGGAACGACAATGAAATGTACCTGTTCCGATCATCTGTTGCATATGCTATGAGGCAGTACTTTTTAAAAGTAAAAAATCAGATGATTCTTTTTGGGGAGGAGGATGTGCGAGTGGCTAATTTGAAACCAAGAATCTCCTTTAATTTCTTTGTCACTGCACCTAAAAATGTGTCTGATATCATTCCTAGAACTGAAGTTGAAAAGGCCATCAGGATGTCCCGGAGCCGTATCAATGATGCTTTCCGTCTGAATGACAACAGCCTAGAGTTTCTGGGGATACAGCCAACACTTGGACCTCCTAACCAGCCCCCTGTTTCCATATGGCTGATTGTTTTTGGAGTTGTGATGGGAGTGATAGTGGTTGGCATTGTCATCCTGATCTTCACTGGGATCAGAGATCGGAAGAAGAAAAATAAAGCAAGAAGTGGAGAAAATCCTTATGCCTCCATCGATATTAGCAAAGGAGAAAATAATCCAGGATTCCAAAACACTGATGATGTTCAGACCTCCTTTTAG
>序列ID 3：huACE2功能域(残基：1-615)蛋白质序列
MSSSSWLLLSLVAVTAAQSTIEEQAKTFLDKFNHEAEDLFYQSSLASWNYNTNITEENVQNMNNAGDKWSAFLKEQSTLAQMYPLQEIQNLTVKLQLQALQQNGSSVLSEDKSKRLNTILNTMSTIYSTGKVCNPDNPQECLLLEPGLNEIMANSLDYNERLWAWESWRSEVGKQLRPLYEEYVVLKNEMARANHYEDYGDYWRGDYEVNGVDGYDYSRGQLIEDVEHTFEEIKPLYEHLHAYVRAKLMNAYPSYISPIGCLPAHLLGDMWGRFWTNLYSLTVPFGQKPNIDVTDAMVDQAWDAQRIFKEAEKFFVSVGLPNMTQGFWENSMLTDPGNVQKAVCHPTAWDLGKGDFRILMCTKVTMDDFLTAHHEMGHIQYDMAYAAQPFLLRNGANEGFHEAVGEIMSLSAATPKHLKSIGLLSPDFQEDNETEINFLLKQALTIVGTLPFTYMLEKWRWMVFKGEIPKDQWMKKWWEMKREIVGVVEPVPHDETYCDPASLFHVSNDYSFIRYYTRTLYQFQFQEALCQAAKHEGPLHKCDISNSTEAGQKLFNMLRLGKSEPWTLALENVVGAKNMNVRPLLNYFEPLFTWLKDQNKNSFVGWSTDWSPYAD
>序列ID 4：huACE2功能域(残基：1-615)DNA序列
ATGTCAAGCTCTTCCTGGCTCCTTCTCAGCCTTGTTGCTGTAACTGCTGCTCAGTCCACCATTGAGGAACAGGCCAAGACATTTTTGGACAAGTTTAACCACGAAGCCGAAGACCTGTTCTATCAAAGTTCACTTGCTTCTTGGAATTATAACACCAATATTACTGAAGAGAATGTCCAAAACATGAATAATGCTGGGGACAAATGGTCTGCCTTTTTAAAGGAACAGTCCACACTTGCCCAAATGTATCCACTACAAGAAATTCAGAATCTCACAGTCAAGCTTCAGCTGCAGGCTCTTCAGCAAAATGGGTCTTCAGTGCTCTCAGAAGACAAGAGCAAACGGTTGAACACAATTCTAAATACAATGAGCACCATCTACAGTACTGGAAAAGTTTGTAACCCAGATAATCCACAAGAATGCTTATTACTTGAACCAGGTTTGAATGAAATAATGGCAAACAGTTTAGACTACAATGAGAGGCTCTGGGCTTGGGAAAGCTGGAGATCTGAGGTCGGCAAGCAGCTGAGGCCATTATATGAAGAGTATGTGGTCTTGAAAAATGAGATGGCAAGAGCAAATCATTATGAGGACTATGGGGATTATTGGAGAGGAGACTATGAAGTAAATGGGGTAGATGGCTATGACTACAGCCGCGGCCAGTTGATTGAAGATGTGGAACATACCTTTGAAGAGATTAAACCATTATATGAACATCTTCATGCCTATGTGAGGGCAAAGTTGATGAATGCCTATCCTTCCTATATCAGTCCAATTGGATGCCTCCCTGCTCATTTGCTTGGTGATATGTGGGGTAGATTTTGGACAAATCTGTACTCTTTGACAGTTCCCTTTGGACAGAAACCAAACATAGATGTTACTGATGCAATGGTGGACCAGGCCTGGGATGCACAGAGAATATTCAAGGAGGCCGAGAAGTTCTTTGTATCTGTTGGTCTTCCTAATATGACTCAAGGATTCTGGGAAAATTCCATGCTAACGGACCCAGGAAATGTTCAGAAAGCAGTCTGCCATCCCACAGCTTGGGACCTGGGGAAGGGCGACTTCAGGATCCTTATGTGCACAAAGGTGACAATGGACGACTTCCTGACAGCTCATCATGAGATGGGGCATATCCAGTATGATATGGCATATGCTGCACAACCTTTTCTGCTAAGAAATGGAGCTAATGAAGGATTCCATGAAGCTGTTGGGGAAATCATGTCACTTTCTGCAGCCACACCTAAGCATTTAAAATCCATTGGTCTTCTGTCACCCGATTTTCAAGAAGACAATGAAACAGAAATAAACTTCCTGCTCAAACAAGCACTCACGATTGTTGGGACTCTGCCATTTACTTACATGTTAGAGAAGTGGAGGTGGATGGTCTTTAAAGGGGAAATTCCCAAAGACCAGTGGATGAAAAAGTGGTGGGAGATGAAGCGAGAGATAGTTGGGGTGGTGGAACCTGTGCCCCATGATGAAACATACTGTGACCCCGCATCTCTGTTCCATGTTTCTAATGATTACTCATTCATTCGATATTACACAAGGACCCTTTACCAATTCCAGTTTCAAGAAGCACTTTGTCAAGCAGCTAAACATGAAGGCCCTCTGCACAAATGTGACATCTCAAACTCTACAGAAGCTGGACAGAAACTGTTCAATATGCTGAGGCTTGGAAAATCAGAACCCTGGACCCTAGCATTGGAAAATGTTGTAGGAGCAAAGAACATGAATGTAAGGCCACTGCTCAACTACTTTGAGCCCTTATTTACCTGGCTGAAAGACCAGAACAAGAATTCTTTTGTGGGATGGAGTACCGACTGGAGTCCATATGCAGAC
>序列ID 5：Fc野生型IgG1 Fc(EU编号216-447)
EPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
>序列ID 6：FC无效版本(EU编号216-447，具有突变C220S、L234A、L235A和K322A)
EPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
>序列ID 7：SI-69R2_huACE2功能域(残基：1-615)-IgG1 Fc(无效)蛋白质序列(EU编号216-447，具有突变C220S、L234A、L235A和K322A)
MSSSSWLLLSLVAVTAAQSTIEEQAKTFLDKFNHEAEDLFYQSSLASWNYNTNITEENVQNMNNAGDKWSAFLKEQSTLAQMYPLQEIQNLTVKLQLQALQQNGSSVLSEDKSKRLNTILNTMSTIYSTGKVCNPDNPQECLLLEPGLNEIMANSLDYNERLWAWESWRSEVGKQLRPLYEEYVVLKNEMARANHYEDYGDYWRGDYEVNGVDGYDYSRGQLIEDVEHTFEEIKPLYEHLHAYVRAKLMNAYPSYISPIGCLPAHLLGDMWGRFWTNLYSLTVPFGQKPNIDVTDAMVDQAWDAQRIFKEAEKFFVSVGLPNMTQGFWENSMLTDPGNVQKAVCHPTAWDLGKGDFRILMCTKVTMDDFLTAHHEMGHIQYDMAYAAQPFLLRNGANEGFHEAVGEIMSLSAATPKHLKSIGLLSPDFQEDNETEINFLLKQALTIVGTLPFTYMLEKWRWMVFKGEIPKDQWMKKWWEMKREIVGVVEPVPHDETYCDPASLFHVSNDYSFIRYYTRTLYQFQFQEALCQAAKHEGPLHKCDISNSTEAGQKLFNMLRLGKSEPWTLALENVVGAKNMNVRPLLNYFEPLFTWLKDQNKNSFVGWSTDWSPYADEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
>序列ID 8：Si-69R2：huACE2功能域(残基：1-615)-IgG1Fc(无效)DNA序列
ATGTCAAGCTCTTCCTGGCTCCTTCTCAGCCTTGTTGCTGTAACTGCTGCTCAGTCCACCATTGAGGAACAGGCCAAGACATTTTTGGACAAGTTTAACCACGAAGCCGAAGACCTGTTCTATCAAAGTTCACTTGCTTCTTGGAATTATAACACCAATATTACTGAAGAGAATGTCCAAAACATGAATAATGCTGGGGACAAATGGTCTGCCTTTTTAAAGGAACAGTCCACACTTGCCCAAATGTATCCACTACAAGAAATTCAGAATCTCACAGTCAAGCTTCAGCTGCAGGCTCTTCAGCAAAATGGGTCTTCAGTGCTCTCAGAAGACAAGAGCAAACGGTTGAACACAATTCTAAATACAATGAGCACCATCTACAGTACTGGAAAAGTTTGTAACCCAGATAATCCACAAGAATGCTTATTACTTGAACCAGGTTTGAATGAAATAATGGCAAACAGTTTAGACTACAATGAGAGGCTCTGGGCTTGGGAAAGCTGGAGATCTGAGGTCGGCAAGCAGCTGAGGCCATTATATGAAGAGTATGTGGTCTTGAAAAATGAGATGGCAAGAGCAAATCATTATGAGGACTATGGGGATTATTGGAGAGGAGACTATGAAGTAAATGGGGTAGATGGCTATGACTACAGCCGCGGCCAGTTGATTGAAGATGTGGAACATACCTTTGAAGAGATTAAACCATTATATGAACATCTTCATGCCTATGTGAGGGCAAAGTTGATGAATGCCTATCCTTCCTATATCAGTCCAATTGGATGCCTCCCTGCTCATTTGCTTGGTGATATGTGGGGTAGATTTTGGACAAATCTGTACTCTTTGACAGTTCCCTTTGGACAGAAACCAAACATAGATGTTACTGATGCAATGGTGGACCAGGCCTGGGATGCACAGAGAATATTCAAGGAGGCCGAGAAGTTCTTTGTATCTGTTGGTCTTCCTAATATGACTCAAGGATTCTGGGAAAATTCCATGCTAACGGACCCAGGAAATGTTCAGAAAGCAGTCTGCCATCCCACAGCTTGGGACCTGGGGAAGGGCGACTTCAGGATCCTTATGTGCACAAAGGTGACAATGGACGACTTCCTGACAGCTCATCATGAGATGGGGCATATCCAGTATGATATGGCATATGCTGCACAACCTTTTCTGCTAAGAAATGGAGCTAATGAAGGATTCCATGAAGCTGTTGGGGAAATCATGTCACTTTCTGCAGCCACACCTAAGCATTTAAAATCCATTGGTCTTCTGTCACCCGATTTTCAAGAAGACAATGAAACAGAAATAAACTTCCTGCTCAAACAAGCACTCACGATTGTTGGGACTCTGCCATTTACTTACATGTTAGAGAAGTGGAGGTGGATGGTCTTTAAAGGGGAAATTCCCAAAGACCAGTGGATGAAAAAGTGGTGGGAGATGAAGCGAGAGATAGTTGGGGTGGTGGAACCTGTGCCCCATGATGAAACATACTGTGACCCCGCATCTCTGTTCCATGTTTCTAATGATTACTCATTCATTCGATATTACACAAGGACCCTTTACCAATTCCAGTTTCAAGAAGCACTTTGTCAAGCAGCTAAACATGAAGGCCCTCTGCACAAATGTGACATCTCAAACTCTACAGAAGCTGGACAGAAACTGTTCAATATGCTGAGGCTTGGAAAATCAGAACCCTGGACCCTAGCATTGGAAAATGTTGTAGGAGCAAAGAACATGAATGTAAGGCCACTGCTCAACTACTTTGAGCCCTTATTTACCTGGCTGAAAGACCAGAACAAGAATTCTTTTGTGGGATGGAGTACCGACTGGAGTCCATATGCAGACGAGCCCAAATCTTCCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCGCGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATAG
>序列ID 9：huACE2功能域(残基：1-615)-IgG4 Fc蛋白质序列
MSSSSWLLLSLVAVTAAQSTIEEQAKTFLDKFNHEAEDLFYQSSLASWNYNTNITEENVQNMNNAGDKWSAFLKEQSTLAQMYPLQEIQNLTVKLQLQALQQNGSSVLSEDKSKRLNTILNTMSTIYSTGKVCNPDNPQECLLLEPGLNEIMANSLDYNERLWAWESWRSEVGKQLRPLYEEYVVLKNEMARANHYEDYGDYWRGDYEVNGVDGYDYSRGQLIEDVEHTFEEIKPLYEHLHAYVRAKLMNAYPSYISPIGCLPAHLLGDMWGRFWTNLYSLTVPFGQKPNIDVTDAMVDQAWDAQRIFKEAEKFFVSVGLPNMTQGFWENSMLTDPGNVQKAVCHPTAWDLGKGDFRILMCTKVTMDDFLTAHHEMGHIQYDMAYAAQPFLLRNGANEGFHEAVGEIMSLSAATPKHLKSIGLLSPDFQEDNETEINFLLKQALTIVGTLPFTYMLEKWRWMVFKGEIPKDQWMKKWWEMKREIVGVVEPVPHDETYCDPASLFHVSNDYSFIRYYTRTLYQFQFQEALCQAAKHEGPLHKCDISNSTEAGQKLFNMLRLGKSEPWTLALENVVGAKNMNVRPLLNYFEPLFTWLKDQNKNSFVGWSTDWSPYADESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK
>序列ID 10：huACE2功能域(残基：1-615)-IgG4 Fc DNA序列
ATGTCAAGCTCTTCCTGGCTCCTTCTCAGCCTTGTTGCTGTAACTGCTGCTCAGTCCACCATTGAGGAACAGGCCAAGACATTTTTGGACAAGTTTAACCACGAAGCCGAAGACCTGTTCTATCAAAGTTCACTTGCTTCTTGGAATTATAACACCAATATTACTGAAGAGAATGTCCAAAACATGAATAATGCTGGGGACAAATGGTCTGCCTTTTTAAAGGAACAGTCCACACTTGCCCAAATGTATCCACTACAAGAAATTCAGAATCTCACAGTCAAGCTTCAGCTGCAGGCTCTTCAGCAAAATGGGTCTTCAGTGCTCTCAGAAGACAAGAGCAAACGGTTGAACACAATTCTAAATACAATGAGCACCATCTACAGTACTGGAAAAGTTTGTAACCCAGATAATCCACAAGAATGCTTATTACTTGAACCAGGTTTGAATGAAATAATGGCAAACAGTTTAGACTACAATGAGAGGCTCTGGGCTTGGGAAAGCTGGAGATCTGAGGTCGGCAAGCAGCTGAGGCCATTATATGAAGAGTATGTGGTCTTGAAAAATGAGATGGCAAGAGCAAATCATTATGAGGACTATGGGGATTATTGGAGAGGAGACTATGAAGTAAATGGGGTAGATGGCTATGACTACAGCCGCGGCCAGTTGATTGAAGATGTGGAACATACCTTTGAAGAGATTAAACCATTATATGAACATCTTCATGCCTATGTGAGGGCAAAGTTGATGAATGCCTATCCTTCCTATATCAGTCCAATTGGATGCCTCCCTGCTCATTTGCTTGGTGATATGTGGGGTAGATTTTGGACAAATCTGTACTCTTTGACAGTTCCCTTTGGACAGAAACCAAACATAGATGTTACTGATGCAATGGTGGACCAGGCCTGGGATGCACAGAGAATATTCAAGGAGGCCGAGAAGTTCTTTGTATCTGTTGGTCTTCCTAATATGACTCAAGGATTCTGGGAAAATTCCATGCTAACGGACCCAGGAAATGTTCAGAAAGCAGTCTGCCATCCCACAGCTTGGGACCTGGGGAAGGGCGACTTCAGGATCCTTATGTGCACAAAGGTGACAATGGACGACTTCCTGACAGCTCATCATGAGATGGGGCATATCCAGTATGATATGGCATATGCTGCACAACCTTTTCTGCTAAGAAATGGAGCTAATGAAGGATTCCATGAAGCTGTTGGGGAAATCATGTCACTTTCTGCAGCCACACCTAAGCATTTAAAATCCATTGGTCTTCTGTCACCCGATTTTCAAGAAGACAATGAAACAGAAATAAACTTCCTGCTCAAACAAGCACTCACGATTGTTGGGACTCTGCCATTTACTTACATGTTAGAGAAGTGGAGGTGGATGGTCTTTAAAGGGGAAATTCCCAAAGACCAGTGGATGAAAAAGTGGTGGGAGATGAAGCGAGAGATAGTTGGGGTGGTGGAACCTGTGCCCCATGATGAAACATACTGTGACCCCGCATCTCTGTTCCATGTTTCTAATGATTACTCATTCATTCGATATTACACAAGGACCCTTTACCAATTCCAGTTTCAAGAAGCACTTTGTCAAGCAGCTAAACATGAAGGCCCTCTGCACAAATGTGACATCTCAAACTCTACAGAAGCTGGACAGAAACTGTTCAATATGCTGAGGCTTGGAAAATCAGAACCCTGGACCCTAGCATTGGAAAATGTTGTAGGAGCAAAGAACATGAATGTAAGGCCACTGCTCAACTACTTTGAGCCCTTATTTACCTGGCTGAAAGACCAGAACAAGAATTCTTTTGTGGGATGGAGTACCGACTGGAGTCCATATGCAGACGAGTCCAAATATGGTCCCCCGTGCCCACCATGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTCCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCTGGGTAAATGA
>序列ID 11：huACE2功能域(残基：1-615)-IgA1 Fc蛋白质序列
MSSSSWLLLSLVAVTAAQSTIEEQAKTFLDKFNHEAEDLFYQSSLASWNYNTNITEENVQNMNNAGDKWSAFLKEQSTLAQMYPLQEIQNLTVKLQLQALQQNGSSVLSEDKSKRLNTILNTMSTIYSTGKVCNPDNPQECLLLEPGLNEIMANSLDYNERLWAWESWRSEVGKQLRPLYEEYVVLKNEMARANHYEDYGDYWRGDYEVNGVDGYDYSRGQLIEDVEHTFEEIKPLYEHLHAYVRAKLMNAYPSYISPIGCLPAHLLGDMWGRFWTNLYSLTVPFGQKPNIDVTDAMVDQAWDAQRIFKEAEKFFVSVGLPNMTQGFWENSMLTDPGNVQKAVCHPTAWDLGKGDFRILMCTKVTMDDFLTAHHEMGHIQYDMAYAAQPFLLRNGANEGFHEAVGEIMSLSAATPKHLKSIGLLSPDFQEDNETEINFLLKQALTIVGTLPFTYMLEKWRWMVFKGEIPKDQWMKKWWEMKREIVGVVEPVPHDETYCDPASLFHVSNDYSFIRYYTRTLYQFQFQEALCQAAKHEGPLHKCDISNSTEAGQKLFNMLRLGKSEPWTLALENVVGAKNMNVRPLLNYFEPLFTWLKDQNKNSFVGWSTDWSPYADSQDVTVPCPVPSTPPTPSPSTPPTPSPSCCHPRLSLHRPALEDLLLGSEANLTCTLTGLRDASGVTFTWTPSSGKSAVQGPPERDLCGCYSVSSVLPGCAEPWNHGKTFTCTAAYPESKTPLTATLSKSGNTFRPEVHLLPPPSEELALNELVTLTCLARGFSPKDVLVRWLQGSQELPREKYLTWASRQEPSQGTTTFAVTSILRVAAEDWKKGDTFSCMVGHEALPLAFTQKTIDRLAGKPTHVNVSVVMAEVDGTCY
>序列ID 12：huACE2功能域(残基：1-615)-IgA1 Fc DNA序列
ATGTCAAGCTCTTCCTGGCTCCTTCTCAGCCTTGTTGCTGTAACTGCTGCTCAGTCCACCATTGAGGAACAGGCCAAGACATTTTTGGACAAGTTTAACCACGAAGCCGAAGACCTGTTCTATCAAAGTTCACTTGCTTCTTGGAATTATAACACCAATATTACTGAAGAGAATGTCCAAAACATGAATAATGCTGGGGACAAATGGTCTGCCTTTTTAAAGGAACAGTCCACACTTGCCCAAATGTATCCACTACAAGAAATTCAGAATCTCACAGTCAAGCTTCAGCTGCAGGCTCTTCAGCAAAATGGGTCTTCAGTGCTCTCAGAAGACAAGAGCAAACGGTTGAACACAATTCTAAATACAATGAGCACCATCTACAGTACTGGAAAAGTTTGTAACCCAGATAATCCACAAGAATGCTTATTACTTGAACCAGGTTTGAATGAAATAATGGCAAACAGTTTAGACTACAATGAGAGGCTCTGGGCTTGGGAAAGCTGGAGATCTGAGGTCGGCAAGCAGCTGAGGCCATTATATGAAGAGTATGTGGTCTTGAAAAATGAGATGGCAAGAGCAAATCATTATGAGGACTATGGGGATTATTGGAGAGGAGACTATGAAGTAAATGGGGTAGATGGCTATGACTACAGCCGCGGCCAGTTGATTGAAGATGTGGAACATACCTTTGAAGAGATTAAACCATTATATGAACATCTTCATGCCTATGTGAGGGCAAAGTTGATGAATGCCTATCCTTCCTATATCAGTCCAATTGGATGCCTCCCTGCTCATTTGCTTGGTGATATGTGGGGTAGATTTTGGACAAATCTGTACTCTTTGACAGTTCCCTTTGGACAGAAACCAAACATAGATGTTACTGATGCAATGGTGGACCAGGCCTGGGATGCACAGAGAATATTCAAGGAGGCCGAGAAGTTCTTTGTATCTGTTGGTCTTCCTAATATGACTCAAGGATTCTGGGAAAATTCCATGCTAACGGACCCAGGAAATGTTCAGAAAGCAGTCTGCCATCCCACAGCTTGGGACCTGGGGAAGGGCGACTTCAGGATCCTTATGTGCACAAAGGTGACAATGGACGACTTCCTGACAGCTCATCATGAGATGGGGCATATCCAGTATGATATGGCATATGCTGCACAACCTTTTCTGCTAAGAAATGGAGCTAATGAAGGATTCCATGAAGCTGTTGGGGAAATCATGTCACTTTCTGCAGCCACACCTAAGCATTTAAAATCCATTGGTCTTCTGTCACCCGATTTTCAAGAAGACAATGAAACAGAAATAAACTTCCTGCTCAAACAAGCACTCACGATTGTTGGGACTCTGCCATTTACTTACATGTTAGAGAAGTGGAGGTGGATGGTCTTTAAAGGGGAAATTCCCAAAGACCAGTGGATGAAAAAGTGGTGGGAGATGAAGCGAGAGATAGTTGGGGTGGTGGAACCTGTGCCCCATGATGAAACATACTGTGACCCCGCATCTCTGTTCCATGTTTCTAATGATTACTCATTCATTCGATATTACACAAGGACCCTTTACCAATTCCAGTTTCAAGAAGCACTTTGTCAAGCAGCTAAACATGAAGGCCCTCTGCACAAATGTGACATCTCAAACTCTACAGAAGCTGGACAGAAACTGTTCAATATGCTGAGGCTTGGAAAATCAGAACCCTGGACCCTAGCATTGGAAAATGTTGTAGGAGCAAAGAACATGAATGTAAGGCCACTGCTCAACTACTTTGAGCCCTTATTTACCTGGCTGAAAGACCAGAACAAGAATTCTTTTGTGGGATGGAGTACCGACTGGAGTCCATATGCAGACAGCCAGGATGTGACTGTGCCCTGCCCAGTTCCCTCAACTCCACCTACCCCATCTCCCTCAACTCCACCTACCCCATCTCCCTCATGCTGCCACCCCCGACTGTCACTGCACCGACCGGCCCTCGAGGACCTGCTCTTAGGTTCAGAAGCGAACCTCACGTGCACACTGACCGGCCTGAGAGATGCCTCAGGTGTCACCTTCACCTGGACGCCCTCAAGTGGGAAGAGCGCTGTTCAAGGACCACCTGAGCGTGACCTCTGTGGCTGCTACAGCGTGTCCAGTGTCCTGCCGGGCTGTGCCGAGCCATGGAACCATGGGAAGACCTTCACTTGCACTGCTGCCTACCCCGAGTCCAAGACCCCGCTAACCGCCACCCTCTCAAAATCCGGAAACACATTCCGGCCCGAGGTCCACCTGCTGCCGCCGCCGTCGGAGGAGCTGGCCCTGAACGAGCTGGTGACGCTGACGTGCCTGGCACGCGGCTTCAGCCCCAAGGACGTGCTGGTTCGCTGGCTGCAGGGGTCACAGGAGCTGCCCCGCGAGAAGTACCTGACTTGGGCATCCCGGCAGGAGCCCAGCCAGGGCACCACCACCTTCGCTGTGACCAGCATACTGCGCGTGGCAGCCGAGGACTGGAAGAAGGGGGACACCTTCTCCTGCATGGTGGGCCACGAGGCCCTGCCGCTGGCCTTCACACAGAAGACCATCGACCGCTTGGCGGGTAAACCCACCCATGTCAATGTGTCTGTTGTCATGGCGGAGGTGGACGGCACCTGCTACTGA
>序列ID 13：huACE2功能域(残基：1-615)-IgA2 Fc蛋白质序列
MSSSSWLLLSLVAVTAAQSTIEEQAKTFLDKFNHEAEDLFYQSSLASWNYNTNITEENVQNMNNAGDKWSAFLKEQSTLAQMYPLQEIQNLTVKLQLQALQQNGSSVLSEDKSKRLNTILNTMSTIYSTGKVCNPDNPQECLLLEPGLNEIMANSLDYNERLWAWESWRSEVGKQLRPLYEEYVVLKNEMARANHYEDYGDYWRGDYEVNGVDGYDYSRGQLIEDVEHTFEEIKPLYEHLHAYVRAKLMNAYPSYISPIGCLPAHLLGDMWGRFWTNLYSLTVPFGQKPNIDVTDAMVDQAWDAQRIFKEAEKFFVSVGLPNMTQGFWENSMLTDPGNVQKAVCHPTAWDLGKGDFRILMCTKVTMDDFLTAHHEMGHIQYDMAYAAQPFLLRNGANEGFHEAVGEIMSLSAATPKHLKSIGLLSPDFQEDNETEINFLLKQALTIVGTLPFTYMLEKWRWMVFKGEIPKDQWMKKWWEMKREIVGVVEPVPHDETYCDPASLFHVSNDYSFIRYYTRTLYQFQFQEALCQAAKHEGPLHKCDISNSTEAGQKLFNMLRLGKSEPWTLALENVVGAKNMNVRPLLNYFEPLFTWLKDQNKNSFVGWSTDWSPYADSQDVTVPCRVPPPPPCCHPRLSLHRPALEDLLLGSEANLTCTLTGLRDASGATFTWTPSSGKSAVQGPPERDLCGCYSVSSVLPGCAQPWNHGETFTCTAAHPELKTPLTANITKSGNTFRPEVHLLPPPSEELALNELVTLTCLARGFSPKDVLVRWLQGSQELPREKYLTWASRQEPSQGTTTYAVTSILRVAAEDWKKGETFSCMVGHEALPLAFTQKTIDRMAGKPTHINVSVVMAEADGTCY
>序列ID 14：huACE2功能域(残基：1-615)-IgA2 Fc DNA序列
ATGTCAAGCTCTTCCTGGCTCCTTCTCAGCCTTGTTGCTGTAACTGCTGCTCAGTCCACCATTGAGGAACAGGCCAAGACATTTTTGGACAAGTTTAACCACGAAGCCGAAGACCTGTTCTATCAAAGTTCACTTGCTTCTTGGAATTATAACACCAATATTACTGAAGAGAATGTCCAAAACATGAATAATGCTGGGGACAAATGGTCTGCCTTTTTAAAGGAACAGTCCACACTTGCCCAAATGTATCCACTACAAGAAATTCAGAATCTCACAGTCAAGCTTCAGCTGCAGGCTCTTCAGCAAAATGGGTCTTCAGTGCTCTCAGAAGACAAGAGCAAACGGTTGAACACAATTCTAAATACAATGAGCACCATCTACAGTACTGGAAAAGTTTGTAACCCAGATAATCCACAAGAATGCTTATTACTTGAACCAGGTTTGAATGAAATAATGGCAAACAGTTTAGACTACAATGAGAGGCTCTGGGCTTGGGAAAGCTGGAGATCTGAGGTCGGCAAGCAGCTGAGGCCATTATATGAAGAGTATGTGGTCTTGAAAAATGAGATGGCAAGAGCAAATCATTATGAGGACTATGGGGATTATTGGAGAGGAGACTATGAAGTAAATGGGGTAGATGGCTATGACTACAGCCGCGGCCAGTTGATTGAAGATGTGGAACATACCTTTGAAGAGATTAAACCATTATATGAACATCTTCATGCCTATGTGAGGGCAAAGTTGATGAATGCCTATCCTTCCTATATCAGTCCAATTGGATGCCTCCCTGCTCATTTGCTTGGTGATATGTGGGGTAGATTTTGGACAAATCTGTACTCTTTGACAGTTCCCTTTGGACAGAAACCAAACATAGATGTTACTGATGCAATGGTGGACCAGGCCTGGGATGCACAGAGAATATTCAAGGAGGCCGAGAAGTTCTTTGTATCTGTTGGTCTTCCTAATATGACTCAAGGATTCTGGGAAAATTCCATGCTAACGGACCCAGGAAATGTTCAGAAAGCAGTCTGCCATCCCACAGCTTGGGACCTGGGGAAGGGCGACTTCAGGATCCTTATGTGCACAAAGGTGACAATGGACGACTTCCTGACAGCTCATCATGAGATGGGGCATATCCAGTATGATATGGCATATGCTGCACAACCTTTTCTGCTAAGAAATGGAGCTAATGAAGGATTCCATGAAGCTGTTGGGGAAATCATGTCACTTTCTGCAGCCACACCTAAGCATTTAAAATCCATTGGTCTTCTGTCACCCGATTTTCAAGAAGACAATGAAACAGAAATAAACTTCCTGCTCAAACAAGCACTCACGATTGTTGGGACTCTGCCATTTACTTACATGTTAGAGAAGTGGAGGTGGATGGTCTTTAAAGGGGAAATTCCCAAAGACCAGTGGATGAAAAAGTGGTGGGAGATGAAGCGAGAGATAGTTGGGGTGGTGGAACCTGTGCCCCATGATGAAACATACTGTGACCCCGCATCTCTGTTCCATGTTTCTAATGATTACTCATTCATTCGATATTACACAAGGACCCTTTACCAATTCCAGTTTCAAGAAGCACTTTGTCAAGCAGCTAAACATGAAGGCCCTCTGCACAAATGTGACATCTCAAACTCTACAGAAGCTGGACAGAAACTGTTCAATATGCTGAGGCTTGGAAAATCAGAACCCTGGACCCTAGCATTGGAAAATGTTGTAGGAGCAAAGAACATGAATGTAAGGCCACTGCTCAACTACTTTGAGCCCTTATTTACCTGGCTGAAAGACCAGAACAAGAATTCTTTTGTGGGATGGAGTACCGACTGGAGTCCATATGCAGACAGCCAGGATGTGACTGTGCCCTGCCGAGTTCCCCCACCTCCCCCATGCTGCCACCCCCGACTGTCGCTGCACCGACCGGCCCTCGAGGACCTGCTCTTAGGTTCAGAAGCGAACCTCACGTGCACACTGACCGGCCTGAGAGATGCCTCTGGTGCCACCTTCACCTGGACGCCCTCAAGTGGGAAGAGCGCTGTTCAAGGACCACCTGAGCGTGACCTCTGTGGCTGCTACAGCGTGTCCAGTGTCCTGCCTGGCTGTGCCCAGCCATGGAACCATGGGGAGACCTTCACCTGCACTGCTGCCCACCCCGAGTTGAAGACCCCACTAACCGCCAACATCACAAAATCCGGAAACACATTCCGGCCCGAGGTCCACCTGCTGCCGCCGCCGTCGGAGGAGCTGGCCCTGAACGAGCTGGTGACGCTGACGTGCCTGGCACGTGGCTTCAGCCCCAAGGATGTGCTGGTTCGCTGGCTGCAGGGGTCACAGGAGCTGCCCCGCGAGAAGTACCTGACTTGGGCATCCCGGCAGGAGCCCAGCCAGGGCACCACCACCTATGCTGTGACCAGCATACTGCGCGTGGCAGCCGAGGACTGGAAGAAGGGGGAAACCTTCTCCTGCATGGTGGGCCACGAGGCCCTGCCGCTGGCCTTCACACAGAAGACCATCGACCGCATGGCGGGTAAACCCACCCATATCAATGTGTCTGTTGTCATGGCGGAGGCGGACGGCACCTGCTACTGA
>序列ID 15：SI-F019_huACE2功能域(残基：18-615)-IgG1Fc(无效)蛋白质序列(具有C220S、L234A、L235A和K322A处的突变，EU编号)
QSTIEEQAKTFLDKFNHEAEDLFYQSSLASWNYNTNITEENVQNMNNAGDKWSAFLKEQSTLAQMYPLQEIQNLTVKLQLQALQQNGSSVLSEDKSKRLNTILNTMSTIYSTGKVCNPDNPQECLLLEPGLNEIMANSLDYNERLWAWESWRSEVGKQLRPLYEEYVVLKNEMARANHYEDYGDYWRGDYEVNGVDGYDYSRGQLIEDVEHTFEEIKPLYEHLHAYVRAKLMNAYPSYISPIGCLPAHLLGDMWGRFWTNLYSLTVPFGQKPNIDVTDAMVDQAWDAQRIFKEAEKFFVSVGLPNMTQGFWENSMLTDPGNVQKAVCHPTAWDLGKGDFRILMCTKVTMDDFLTAHHEMGHIQYDMAYAAQPFLLRNGANEGFHEAVGEIMSLSAATPKHLKSIGLLSPDFQEDNETEINFLLKQALTIVGTLPFTYMLEKWRWMVFKGEIPKDQWMKKWWEMKREIVGVVEPVPHDETYCDPASLFHVSNDYSFIRYYTRTLYQFQFQEALCQAAKHEGPLHKCDISNSTEAGQKLFNMLRLGKSEPWTLALENVVGAKNMNVRPLLNYFEPLFTWLKDQNKNSFVGWSTDWSPYADEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
>序列ID 16：huACE2功能域(残基：18-615)-IgG4 Fc蛋白质序列
QSTIEEQAKTFLDKFNHEAEDLFYQSSLASWNYNTNITEENVQNMNNAGDKWSAFLKEQSTLAQMYPLQEIQNLTVKLQLQALQQNGSSVLSEDKSKRLNTILNTMSTIYSTGKVCNPDNPQECLLLEPGLNEIMANSLDYNERLWAWESWRSEVGKQLRPLYEEYVVLKNEMARANHYEDYGDYWRGDYEVNGVDGYDYSRGQLIEDVEHTFEEIKPLYEHLHAYVRAKLMNAYPSYISPIGCLPAHLLGDMWGRFWTNLYSLTVPFGQKPNIDVTDAMVDQAWDAQRIFKEAEKFFVSVGLPNMTQGFWENSMLTDPGNVQKAVCHPTAWDLGKGDFRILMCTKVTMDDFLTAHHEMGHIQYDMAYAAQPFLLRNGANEGFHEAVGEIMSLSAATPKHLKSIGLLSPDFQEDNETEINFLLKQALTIVGTLPFTYMLEKWRWMVFKGEIPKDQWMKKWWEMKREIVGVVEPVPHDETYCDPASLFHVSNDYSFIRYYTRTLYQFQFQEALCQAAKHEGPLHKCDISNSTEAGQKLFNMLRLGKSEPWTLALENVVGAKNMNVRPLLNYFEPLFTWLKDQNKNSFVGWSTDWSPYADESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK
>序列ID 17：huACE2功能域(残基：18-615)-IgA1 Fc蛋白质序列
QSTIEEQAKTFLDKFNHEAEDLFYQSSLASWNYNTNITEENVQNMNNAGDKWSAFLKEQSTLAQMYPLQEIQNLTVKLQLQALQQNGSSVLSEDKSKRLNTILNTMSTIYSTGKVCNPDNPQECLLLEPGLNEIMANSLDYNERLWAWESWRSEVGKQLRPLYEEYVVLKNEMARANHYEDYGDYWRGDYEVNGVDGYDYSRGQLIEDVEHTFEEIKPLYEHLHAYVRAKLMNAYPSYISPIGCLPAHLLGDMWGRFWTNLYSLTVPFGQKPNIDVTDAMVDQAWDAQRIFKEAEKFFVSVGLPNMTQGFWENSMLTDPGNVQKAVCHPTAWDLGKGDFRILMCTKVTMDDFLTAHHEMGHIQYDMAYAAQPFLLRNGANEGFHEAVGEIMSLSAATPKHLKSIGLLSPDFQEDNETEINFLLKQALTIVGTLPFTYMLEKWRWMVFKGEIPKDQWMKKWWEMKREIVGVVEPVPHDETYCDPASLFHVSNDYSFIRYYTRTLYQFQFQEALCQAAKHEGPLHKCDISNSTEAGQKLFNMLRLGKSEPWTLALENVVGAKNMNVRPLLNYFEPLFTWLKDQNKNSFVGWSTDWSPYADSQDVTVPCPVPSTPPTPSPSTPPTPSPSCCHPRLSLHRPALEDLLLGSEANLTCTLTGLRDASGVTFTWTPSSGKSAVQGPPERDLCGCYSVSSVLPGCAEPWNHGKTFTCTAAYPESKTPLTATLSKSGNTFRPEVHLLPPPSEELALNELVTLTCLARGFSPKDVLVRWLQGSQELPREKYLTWASRQEPSQGTTTFAVTSILRVAAEDWKKGDTFSCMVGHEALPLAFTQKTIDRLAGKPTHVNVSVVMAEVDGTCY
>序列ID 18：huACE2功能域(残基：18-615)-IgA2 Fc蛋白质序列
QSTIEEQAKTFLDKFNHEAEDLFYQSSLASWNYNTNITEENVQNMNNAGDKWSAFLKEQSTLAQMYPLQEIQNLTVKLQLQALQQNGSSVLSEDKSKRLNTILNTMSTIYSTGKVCNPDNPQECLLLEPGLNEIMANSLDYNERLWAWESWRSEVGKQLRPLYEEYVVLKNEMARANHYEDYGDYWRGDYEVNGVDGYDYSRGQLIEDVEHTFEEIKPLYEHLHAYVRAKLMNAYPSYISPIGCLPAHLLGDMWGRFWTNLYSLTVPFGQKPNIDVTDAMVDQAWDAQRIFKEAEKFFVSVGLPNMTQGFWENSMLTDPGNVQKAVCHPTAWDLGKGDFRILMCTKVTMDDFLTAHHEMGHIQYDMAYAAQPFLLRNGANEGFHEAVGEIMSLSAATPKHLKSIGLLSPDFQEDNETEINFLLKQALTIVGTLPFTYMLEKWRWMVFKGEIPKDQWMKKWWEMKREIVGVVEPVPHDETYCDPASLFHVSNDYSFIRYYTRTLYQFQFQEALCQAAKHEGPLHKCDISNSTEAGQKLFNMLRLGKSEPWTLALENVVGAKNMNVRPLLNYFEPLFTWLKDQNKNSFVGWSTDWSPYADSQDVTVPCRVPPPPPCCHPRLSLHRPALEDLLLGSEANLTCTLTGLRDASGATFTWTPSSGKSAVQGPPERDLCGCYSVSSVLPGCAQPWNHGETFTCTAAHPELKTPLTANITKSGNTFRPEVHLLPPPSEELALNELVTLTCLARGFSPKDVLVRWLQGSQELPREKYLTWASRQEPSQGTTTYAVTSILRVAAEDWKKGETFSCMVGHEALPLAFTQKTIDRMAGKPTHINVSVVMAEADGTCY
>序列ID 19：SI-69R3_human ACE2-ECD-1-615-Fc-w2(EU编号216-447)-蛋白质序列
MSSSSWLLLSLVAVTAAQSTIEEQAKTFLDKFNHEAEDLFYQSSLASWNYNTNITEENVQNMNNAGDKWSAFLKEQSTLAQMYPLQEIQNLTVKLQLQALQQNGSSVLSEDKSKRLNTILNTMSTIYSTGKVCNPDNPQECLLLEPGLNEIMANSLDYNERLWAWESWRSEVGKQLRPLYEEYVVLKNEMARANHYEDYGDYWRGDYEVNGVDGYDYSRGQLIEDVEHTFEEIKPLYEHLHAYVRAKLMNAYPSYISPIGCLPAHLLGDMWGRFWTNLYSLTVPFGQKPNIDVTDAMVDQAWDAQRIFKEAEKFFVSVGLPNMTQGFWENSMLTDPGNVQKAVCHPTAWDLGKGDFRILMCTKVTMDDFLTAHHEMGHIQYDMAYAAQPFLLRNGANEGFHEAVGEIMSLSAATPKHLKSIGLLSPDFQEDNETEINFLLKQALTIVGTLPFTYMLEKWRWMVFKGEIPKDQWMKKWWEMKREIVGVVEPVPHDETYCDPASLFHVSNDYSFIRYYTRTLYQFQFQEALCQAAKHEGPLHKCDISNSTEAGQKLFNMLRLGKSEPWTLALENVVGAKNMNVRPLLNYFEPLFTWLKDQNKNSFVGWSTDWSPYADEPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
>序列ID 20：SI-69R3_human ACE2-ECD-1-615-Fc-w2-DNA序列
ATGTCAAGCTCTTCCTGGCTCCTTCTCAGCCTTGTTGCTGTAACTGCTGCTCAGTCCACCATTGAGGAACAGGCCAAGACATTTTTGGACAAGTTTAACCACGAAGCCGAAGACCTGTTCTATCAAAGTTCACTTGCTTCTTGGAATTATAACACCAATATTACTGAAGAGAATGTCCAAAACATGAATAATGCTGGGGACAAATGGTCTGCCTTTTTAAAGGAACAGTCCACACTTGCCCAAATGTATCCACTACAAGAAATTCAGAATCTCACAGTCAAGCTCCAGCTGCAGGCTCTTCAGCAAAATGGGTCTTCAGTGCTCTCAGAAGACAAGAGCAAACGGTTGAACACAATTCTAAATACAATGAGCACCATCTACAGTACTGGAAAAGTTTGTAACCCAGATAATCCACAAGAATGCTTATTACTTGAACCAGGTTTGAATGAAATAATGGCAAACAGTTTAGACTACAATGAGAGGCTCTGGGCTTGGGAAAGCTGGAGATCTGAGGTCGGCAAGCAGCTGAGGCCATTATATGAAGAGTATGTGGTCTTGAAAAATGAGATGGCAAGAGCAAATCATTATGAGGACTATGGGGATTATTGGAGAGGAGACTATGAAGTAAATGGGGTAGATGGCTATGACTACAGCCGCGGCCAGTTGATTGAAGATGTGGAACATACCTTTGAAGAGATTAAACCATTATATGAACATCTTCATGCCTATGTGAGGGCAAAGTTGATGAATGCCTATCCTTCCTATATCAGTCCAATTGGATGCCTCCCTGCTCATTTGCTTGGTGATATGTGGGGTAGATTTTGGACAAATCTGTACTCTTTGACAGTTCCCTTTGGACAGAAACCAAACATAGATGTTACTGATGCAATGGTGGACCAGGCCTGGGATGCACAGAGAATATTCAAGGAGGCCGAGAAGTTCTTTGTATCTGTTGGTCTTCCTAATATGACTCAAGGATTCTGGGAAAATTCCATGCTAACGGACCCAGGAAATGTTCAGAAAGCAGTCTGCCATCCCACAGCTTGGGACCTGGGGAAGGGCGACTTCAGGATCCTTATGTGCACAAAGGTGACAATGGACGACTTCCTGACAGCTCATCATGAGATGGGGCATATCCAGTATGATATGGCATATGCTGCACAACCTTTTCTGCTAAGAAATGGAGCTAATGAAGGATTCCATGAAGCTGTTGGGGAAATCATGTCACTTTCTGCAGCCACACCTAAGCATTTAAAATCCATTGGTCTTCTGTCACCCGATTTTCAAGAAGACAATGAAACAGAAATAAACTTCCTGCTCAAACAAGCACTCACGATTGTTGGGACTCTGCCATTTACTTACATGTTAGAGAAGTGGAGGTGGATGGTCTTTAAAGGGGAAATTCCCAAAGACCAGTGGATGAAAAAGTGGTGGGAGATGAAGCGAGAGATAGTTGGGGTGGTGGAACCTGTGCCCCATGATGAAACATACTGTGACCCCGCATCTCTGTTCCATGTTTCTAATGATTACTCATTCATTCGATATTACACAAGGACCCTTTACCAATTCCAGTTTCAAGAAGCACTTTGTCAAGCAGCTAAACATGAAGGCCCTCTGCACAAATGTGACATCTCAAACTCTACAGAAGCTGGACAGAAACTGTTCAATATGCTGAGGCTTGGAAAATCAGAACCCTGGACCCTAGCATTGGAAAATGTTGTAGGAGCAAAGAACATGAATGTAAGGCCACTGCTCAACTACTTTGAGCCCTTATTTACCTGGCTGAAAGACCAGAACAAGAATTCTTTTGTGGGATGGAGTACCGACTGGAGTCCATATGCAGACGAGCCCAAATCTTCCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATAG
>序列ID21：SI-69R4-human ACE2-ECD-1-740(TMPRSS2蛋白酶切割位点)-Fc-w2(EU编号216-447)-蛋白质序列
MSSSSWLLLSLVAVTAAQSTIEEQAKTFLDKFNHEAEDLFYQSSLASWNYNTNITEENVQNMNNAGDKWSAFLKEQSTLAQMYPLQEIQNLTVKLQLQALQQNGSSVLSEDKSKRLNTILNTMSTIYSTGKVCNPDNPQECLLLEPGLNEIMANSLDYNERLWAWESWRSEVGKQLRPLYEEYVVLKNEMARANHYEDYGDYWRGDYEVNGVDGYDYSRGQLIEDVEHTFEEIKPLYEHLHAYVRAKLMNAYPSYISPIGCLPAHLLGDMWGRFWTNLYSLTVPFGQKPNIDVTDAMVDQAWDAQRIFKEAEKFFVSVGLPNMTQGFWENSMLTDPGNVQKAVCHPTAWDLGKGDFRILMCTKVTMDDFLTAHHEMGHIQYDMAYAAQPFLLRNGANEGFHEAVGEIMSLSAATPKHLKSIGLLSPDFQEDNETEINFLLKQALTIVGTLPFTYMLEKWRWMVFKGEIPKDQWMKKWWEMKREIVGVVEPVPHDETYCDPASLFHVSNDYSFIRYYTRTLYQFQFQEALCQAAKHEGPLHKCDISNSTEAGQKLFNMLRLGKSEPWTLALENVVGAKNMNVRPLLNYFEPLFTWLKDQNKNSFVGWSTDWSPYADQSIKVRISLKSALGDKAYEWNDNEMYLFRSSVAYAMRQYFLKVKNQMILFGEEDVRVANLKPRISFNFFVTAPKNVSDIIPRTEVEKAIRMSRSRINDAFRLNDNSLEFLGIQPTLGPPNQPPVSEPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
>序列ID 22：SI-69R4_human ACE2-ECD-1-740-Fc-w2-DNA序列
ATGTCAAGCTCTTCCTGGCTCCTTCTCAGCCTTGTTGCTGTAACTGCTGCTCAGTCCACCATTGAGGAACAGGCCAAGACATTTTTGGACAAGTTTAACCACGAAGCCGAAGACCTGTTCTATCAAAGTTCACTTGCTTCTTGGAATTATAACACCAATATTACTGAAGAGAATGTCCAAAACATGAATAATGCTGGGGACAAATGGTCTGCCTTTTTAAAGGAACAGTCCACACTTGCCCAAATGTATCCACTACAAGAAATTCAGAATCTCACAGTCAAGCTCCAGCTGCAGGCTCTTCAGCAAAATGGGTCTTCAGTGCTCTCAGAAGACAAGAGCAAACGGTTGAACACAATTCTAAATACAATGAGCACCATCTACAGTACTGGAAAAGTTTGTAACCCAGATAATCCACAAGAATGCTTATTACTTGAACCAGGTTTGAATGAAATAATGGCAAACAGTTTAGACTACAATGAGAGGCTCTGGGCTTGGGAAAGCTGGAGATCTGAGGTCGGCAAGCAGCTGAGGCCATTATATGAAGAGTATGTGGTCTTGAAAAATGAGATGGCAAGAGCAAATCATTATGAGGACTATGGGGATTATTGGAGAGGAGACTATGAAGTAAATGGGGTAGATGGCTATGACTACAGCCGCGGCCAGTTGATTGAAGATGTGGAACATACCTTTGAAGAGATTAAACCATTATATGAACATCTTCATGCCTATGTGAGGGCAAAGTTGATGAATGCCTATCCTTCCTATATCAGTCCAATTGGATGCCTCCCTGCTCATTTGCTTGGTGATATGTGGGGTAGATTTTGGACAAATCTGTACTCTTTGACAGTTCCCTTTGGACAGAAACCAAACATAGATGTTACTGATGCAATGGTGGACCAGGCCTGGGATGCACAGAGAATATTCAAGGAGGCCGAGAAGTTCTTTGTATCTGTTGGTCTTCCTAATATGACTCAAGGATTCTGGGAAAATTCCATGCTAACGGACCCAGGAAATGTTCAGAAAGCAGTCTGCCATCCCACAGCTTGGGACCTGGGGAAGGGCGACTTCAGGATCCTTATGTGCACAAAGGTGACAATGGACGACTTCCTGACAGCTCATCATGAGATGGGGCATATCCAGTATGATATGGCATATGCTGCACAACCTTTTCTGCTAAGAAATGGAGCTAATGAAGGATTCCATGAAGCTGTTGGGGAAATCATGTCACTTTCTGCAGCCACACCTAAGCATTTAAAATCCATTGGTCTTCTGTCACCCGATTTTCAAGAAGACAATGAAACAGAAATAAACTTCCTGCTCAAACAAGCACTCACGATTGTTGGGACTCTGCCATTTACTTACATGTTAGAGAAGTGGAGGTGGATGGTCTTTAAAGGGGAAATTCCCAAAGACCAGTGGATGAAAAAGTGGTGGGAGATGAAGCGAGAGATAGTTGGGGTGGTGGAACCTGTGCCCCATGATGAAACATACTGTGACCCCGCATCTCTGTTCCATGTTTCTAATGATTACTCATTCATTCGATATTACACAAGGACCCTTTACCAATTCCAGTTTCAAGAAGCACTTTGTCAAGCAGCTAAACATGAAGGCCCTCTGCACAAATGTGACATCTCAAACTCTACAGAAGCTGGACAGAAACTGTTCAATATGCTGAGGCTTGGAAAATCAGAACCCTGGACCCTAGCATTGGAAAATGTTGTAGGAGCAAAGAACATGAATGTAAGGCCACTGCTCAACTACTTTGAGCCCTTATTTACCTGGCTGAAAGACCAGAACAAGAATTCTTTTGTGGGATGGAGTACCGACTGGAGTCCATATGCAGACCAAAGCATCAAAGTGAGGATAAGCCTAAAATCAGCTCTTGGAGATAAAGCATATGAATGGAACGACAATGAAATGTACCTGTTCCGATCATCTGTTGCATATGCTATGAGGCAGTACTTTTTAAAAGTAAAAAATCAGATGATTCTTTTTGGGGAGGAGGATGTGCGAGTGGCTAATTTGAAACCAAGAATCTCCTTTAATTTCTTTGTCACTGCACCTAAAAATGTGTCTGATATCATTCCTAGAACTGAAGTTGAAAAGGCCATCAGGATGTCCCGGAGCCGTATCAATGATGCTTTCCGTCTGAATGACAACAGCCTAGAGTTTCTGGGGATACAGCCAACACTTGGACCTCCTAACCAGCCCCCTGTTTCCGAGCCCAAATCTTCCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATAG
>序列ID 23：SI-69R1_huACE2功能域(残基：1-615)-6XHis蛋白质序列
MSSSSWLLLSLVAVTAAQSTIEEQAKTFLDKFNHEAEDLFYQSSLASWNYNTNITEENVQNMNNAGDKWSAFLKEQSTLAQMYPLQEIQNLTVKLQLQALQQNGSSVLSEDKSKRLNTILNTMSTIYSTGKVCNPDNPQECLLLEPGLNEIMANSLDYNERLWAWESWRSEVGKQLRPLYEEYVVLKNEMARANHYEDYGDYWRGDYEVNGVDGYDYSRGQLIEDVEHTFEEIKPLYEHLHAYVRAKLMNAYPSYISPIGCLPAHLLGDMWGRFWTNLYSLTVPFGQKPNIDVTDAMVDQAWDAQRIFKEAEKFFVSVGLPNMTQGFWENSMLTDPGNVQKAVCHPTAWDLGKGDFRILMCTKVTMDDFLTAHHEMGHIQYDMAYAAQPFLLRNGANEGFHEAVGEIMSLSAATPKHLKSIGLLSPDFQEDNETEINFLLKQALTIVGTLPFTYMLEKWRWMVFKGEIPKDQWMKKWWEMKREIVGVVEPVPHDETYCDPASLFHVSNDYSFIRYYTRTLYQFQFQEALCQAAKHEGPLHKCDISNSTEAGQKLFNMLRLGKSEPWTLALENVVGAKNMNVRPLLNYFEPLFTWLKDQNKNSFVGWSTDWSPYADHHHHHH
>序列ID 24：SI-69R1_huACE2功能域(残基：1-615)-6XHis DNA序列
ATGTCAAGCTCTTCCTGGCTCCTTCTCAGCCTTGTTGCTGTAACTGCTGCTCAGTCCACCATTGAGGAACAGGCCAAGACATTTTTGGACAAGTTTAACCACGAAGCCGAAGACCTGTTCTATCAAAGTTCACTTGCTTCTTGGAATTATAACACCAATATTACTGAAGAGAATGTCCAAAACATGAATAATGCTGGGGACAAATGGTCTGCCTTTTTAAAGGAACAGTCCACACTTGCCCAAATGTATCCACTACAAGAAATTCAGAATCTCACAGTCAAGCTTCAGCTGCAGGCTCTTCAGCAAAATGGGTCTTCAGTGCTCTCAGAAGACAAGAGCAAACGGTTGAACACAATTCTAAATACAATGAGCACCATCTACAGTACTGGAAAAGTTTGTAACCCAGATAATCCACAAGAATGCTTATTACTTGAACCAGGTTTGAATGAAATAATGGCAAACAGTTTAGACTACAATGAGAGGCTCTGGGCTTGGGAAAGCTGGAGATCTGAGGTCGGCAAGCAGCTGAGGCCATTATATGAAGAGTATGTGGTCTTGAAAAATGAGATGGCAAGAGCAAATCATTATGAGGACTATGGGGATTATTGGAGAGGAGACTATGAAGTAAATGGGGTAGATGGCTATGACTACAGCCGCGGCCAGTTGATTGAAGATGTGGAACATACCTTTGAAGAGATTAAACCATTATATGAACATCTTCATGCCTATGTGAGGGCAAAGTTGATGAATGCCTATCCTTCCTATATCAGTCCAATTGGATGCCTCCCTGCTCATTTGCTTGGTGATATGTGGGGTAGATTTTGGACAAATCTGTACTCTTTGACAGTTCCCTTTGGACAGAAACCAAACATAGATGTTACTGATGCAATGGTGGACCAGGCCTGGGATGCACAGAGAATATTCAAGGAGGCCGAGAAGTTCTTTGTATCTGTTGGTCTTCCTAATATGACTCAAGGATTCTGGGAAAATTCCATGCTAACGGACCCAGGAAATGTTCAGAAAGCAGTCTGCCATCCCACAGCTTGGGACCTGGGGAAGGGCGACTTCAGGATCCTTATGTGCACAAAGGTGACAATGGACGACTTCCTGACAGCTCATCATGAGATGGGGCATATCCAGTATGATATGGCATATGCTGCACAACCTTTTCTGCTAAGAAATGGAGCTAATGAAGGATTCCATGAAGCTGTTGGGGAAATCATGTCACTTTCTGCAGCCACACCTAAGCATTTAAAATCCATTGGTCTTCTGTCACCCGATTTTCAAGAAGACAATGAAACAGAAATAAACTTCCTGCTCAAACAAGCACTCACGATTGTTGGGACTCTGCCATTTACTTACATGTTAGAGAAGTGGAGGTGGATGGTCTTTAAAGGGGAAATTCCCAAAGACCAGTGGATGAAAAAGTGGTGGGAGATGAAGCGAGAGATAGTTGGGGTGGTGGAACCTGTGCCCCATGATGAAACATACTGTGACCCCGCATCTCTGTTCCATGTTTCTAATGATTACTCATTCATTCGATATTACACAAGGACCCTTTACCAATTCCAGTTTCAAGAAGCACTTTGTCAAGCAGCTAAACATGAAGGCCCTCTGCACAAATGTGACATCTCAAACTCTACAGAAGCTGGACAGAAACTGTTCAATATGCTGAGGCTTGGAAAATCAGAACCCTGGACCCTAGCATTGGAAAATGTTGTAGGAGCAAAGAACATGAATGTAAGGCCACTGCTCAACTACTTTGAGCCCTTATTTACCTGGCTGAAAGACCAGAACAAGAATTCTTTTGTGGGATGGAGTACCGACTGGAGTCCATATGCAGACCATCATCACCATCACCAC
>序列ID 25：SI-69R10_Human TMPRSS2蛋白，His-标签(106-492)-蛋白质序列
MYRMQLLSCIALSLALVTNSWKFMGSKCSNSGIECDSSGTCINPSNWCDGVSHCPGGEDENRCVRLYGPNFILQVYSSQRKSWHPVCQDDWNENYGRAACRDMGYKNNFYSSQGIVDDSGSTSFMKLNTSAGNVDIYKKLYHSDACSSKAVVSLRCIACGVNLNSSRQSRIVGGESALPGAWPWQVSLHVQNVHVCGGSIITPEWIVTAAHCVEKPLNNPWHWTAFAGILRQSFMFYGAGYQVEKVISHPNYDSKTKNNDIALMKLQKPLTFNDLVKPVCLPNPGMMLQPEQLCWISGWGATEEKGKTSEVLNAAKVLLIETQRCNSRYVYDNLITPAMICAGFLQGNVDSCQGDSGGPLVTSKNNIWWLIGDTSWGSGCAKAYRPGVYGNVMVFTDWIYRQMRADGHHHHHH
>序列ID 26：SI-69R10_Human TMPRSS2蛋白，His-标签(106-492)-DNA序列
ATGTACAGGATGCAACTCCTGTCTTGCATTGCACTAAGTCTTGCACTTGTCACCAATTCGTGGAAGTTTATGGGTTCTAAATGCTCTAATAGCGGGATAGAATGTGACAGTAGTGGCACTTGCATTAACCCTTCAAACTGGTGTGATGGGGTAAGCCATTGCCCCGGGGGGGAAGATGAAAATAGATGTGTTAGGCTCTACGGTCCCAACTTTATACTCCAGGTATATTCAAGTCAACGCAAATCATGGCATCCAGTGTGTCAAGACGACTGGAACGAAAACTATGGACGCGCTGCATGTCGAGATATGGGATATAAGAATAACTTCTATAGTTCACAGGGAATCGTAGATGACTCTGGATCTACTAGTTTCATGAAACTGAACACCTCTGCCGGAAACGTAGATATATATAAAAAGCTTTACCACTCCGACGCTTGTAGCTCTAAGGCCGTAGTTAGCCTCAGATGCATCGCCTGCGGAGTAAACCTCAATTCATCTCGCCAGAGTAGGATCGTTGGCGGGGAAAGCGCCCTCCCAGGCGCTTGGCCTTGGCAAGTTTCCCTTCATGTCCAGAATGTTCATGTATGTGGCGGGTCTATAATCACCCCAGAATGGATCGTCACAGCTGCCCACTGCGTGGAGAAACCCCTCAACAATCCTTGGCATTGGACCGCATTTGCCGGAATACTGAGACAATCATTTATGTTCTATGGAGCCGGGTACCAAGTCGAAAAGGTCATTTCCCATCCCAATTATGATTCCAAAACCAAAAACAATGACATAGCCTTGATGAAACTCCAGAAGCCTTTGACATTTAATGACCTGGTCAAACCAGTGTGCCTCCCAAATCCTGGAATGATGTTGCAGCCTGAACAGTTGTGCTGGATCAGCGGTTGGGGTGCTACCGAGGAGAAGGGTAAGACAAGCGAGGTCCTTAACGCTGCAAAGGTTTTGCTGATAGAAACACAGAGATGTAACAGCCGCTATGTGTACGATAACCTGATCACCCCAGCTATGATTTGCGCCGGGTTTTTGCAAGGTAACGTCGATTCTTGCCAAGGTGACTCAGGCGGCCCTCTTGTTACATCAAAGAACAATATATGGTGGCTTATCGGCGATACATCATGGGGTTCTGGATGTGCTAAAGCCTATCGCCCAGGGGTGTATGGCAATGTAATGGTGTTTACAGACTGGATCTATAGGCAGATGCGGGCTGACGGTCACCATCATCACCATCACTGA
>序列ID 27：IgJ链
MKNHLLFWGVLAVFIKAVHVKAQEDERIVLVDNKCKCARITSRIIRSSEDPNEDIVERNIRIIVPLNNRENISDPTSPLRTRFVYHLSDLCKKCDPTEVELDNQIVTATQSNICDEDSATETCYTYDRNKCYTAVVPLVYGGETKMVETALTPDACYPD
>序列ID 28：分泌成分
KSPIFGPEEVNSVEGNSVSITCYYPPTSVNRHTRKYWCRQGARGGCITLISSEGYVSSKYAGRANLTNFPENGTFVVNIAQLSQDDSGRYKCGLGINSRGLSFDVSLEVSQGPGLLNDTKVYTVDLGRTVTINCPFKTENAQKRKSLYKQIGLYPVLVIDSSGYVNPNYTGRIRLDIQGTGQLLFSVVINQLRLSDAGQYLCQAGDDSNSNKKNADLQVLKPEPELVYEDLRGSVTFHCALGPEVANVAKFLCRQSSGENCDVVVNTLGKRAPAFEGRILLNPQDKDGSFSVVITGLRKEDAGRYLCGAHSDGQLQEGSPIQAWQLFVNEESTIPRSPTVVKGVAGGSVAVLCPYNRKESKSIKYWCLWEGAQNGRCPLLVDSEGWVKAQYEGRLSLLEEPGNGTFTVILNQLTSRDAGFYWCLTNGDTLWRTTVEIKIIEGEPNLKVPGNVTAVLGETLKVPCHFPCKFSSYEKYWCKWNNTGCQALPSQDEGPSKAFVNCDENSRLVSLTLNLVTRADEGWYWCGVKQGHFYGETAAVYVAVEERKAAGSRDVSLAKADAAPDEKVLDSGFREIENKAIQDPR
序列表
<110> 西雅图免疫公司
四川百利药业有限责任公司
<120> 重组ACE2-FC融合分子及其制备和使用方法
<130> SIBA202PCT
<141> 2021-02-10
<160> 28
<170> SIPOSequenceListing 1.0
<210> 1
<211> 805
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 1
Met Ser Ser Ser Ser Trp Leu Leu Leu Ser Leu Val Ala Val Thr Ala
1 5 10 15
Ala Gln Ser Thr Ile Glu Glu Gln Ala Lys Thr Phe Leu Asp Lys Phe
20 25 30
Asn His Glu Ala Glu Asp Leu Phe Tyr Gln Ser Ser Leu Ala Ser Trp
35 40 45
Asn Tyr Asn Thr Asn Ile Thr Glu Glu Asn Val Gln Asn Met Asn Asn
50 55 60
Ala Gly Asp Lys Trp Ser Ala Phe Leu Lys Glu Gln Ser Thr Leu Ala
65 70 75 80
Gln Met Tyr Pro Leu Gln Glu Ile Gln Asn Leu Thr Val Lys Leu Gln
85 90 95
Leu Gln Ala Leu Gln Gln Asn Gly Ser Ser Val Leu Ser Glu Asp Lys
100 105 110
Ser Lys Arg Leu Asn Thr Ile Leu Asn Thr Met Ser Thr Ile Tyr Ser
115 120 125
Thr Gly Lys Val Cys Asn Pro Asp Asn Pro Gln Glu Cys Leu Leu Leu
130 135 140
Glu Pro Gly Leu Asn Glu Ile Met Ala Asn Ser Leu Asp Tyr Asn Glu
145 150 155 160
Arg Leu Trp Ala Trp Glu Ser Trp Arg Ser Glu Val Gly Lys Gln Leu
165 170 175
Arg Pro Leu Tyr Glu Glu Tyr Val Val Leu Lys Asn Glu Met Ala Arg
180 185 190
Ala Asn His Tyr Glu Asp Tyr Gly Asp Tyr Trp Arg Gly Asp Tyr Glu
195 200 205
Val Asn Gly Val Asp Gly Tyr Asp Tyr Ser Arg Gly Gln Leu Ile Glu
210 215 220
Asp Val Glu His Thr Phe Glu Glu Ile Lys Pro Leu Tyr Glu His Leu
225 230 235 240
His Ala Tyr Val Arg Ala Lys Leu Met Asn Ala Tyr Pro Ser Tyr Ile
245 250 255
Ser Pro Ile Gly Cys Leu Pro Ala His Leu Leu Gly Asp Met Trp Gly
260 265 270
Arg Phe Trp Thr Asn Leu Tyr Ser Leu Thr Val Pro Phe Gly Gln Lys
275 280 285
Pro Asn Ile Asp Val Thr Asp Ala Met Val Asp Gln Ala Trp Asp Ala
290 295 300
Gln Arg Ile Phe Lys Glu Ala Glu Lys Phe Phe Val Ser Val Gly Leu
305 310 315 320
Pro Asn Met Thr Gln Gly Phe Trp Glu Asn Ser Met Leu Thr Asp Pro
325 330 335
Gly Asn Val Gln Lys Ala Val Cys His Pro Thr Ala Trp Asp Leu Gly
340 345 350
Lys Gly Asp Phe Arg Ile Leu Met Cys Thr Lys Val Thr Met Asp Asp
355 360 365
Phe Leu Thr Ala His His Glu Met Gly His Ile Gln Tyr Asp Met Ala
370 375 380
Tyr Ala Ala Gln Pro Phe Leu Leu Arg Asn Gly Ala Asn Glu Gly Phe
385 390 395 400
His Glu Ala Val Gly Glu Ile Met Ser Leu Ser Ala Ala Thr Pro Lys
405 410 415
His Leu Lys Ser Ile Gly Leu Leu Ser Pro Asp Phe Gln Glu Asp Asn
420 425 430
Glu Thr Glu Ile Asn Phe Leu Leu Lys Gln Ala Leu Thr Ile Val Gly
435 440 445
Thr Leu Pro Phe Thr Tyr Met Leu Glu Lys Trp Arg Trp Met Val Phe
450 455 460
Lys Gly Glu Ile Pro Lys Asp Gln Trp Met Lys Lys Trp Trp Glu Met
465 470 475 480
Lys Arg Glu Ile Val Gly Val Val Glu Pro Val Pro His Asp Glu Thr
485 490 495
Tyr Cys Asp Pro Ala Ser Leu Phe His Val Ser Asn Asp Tyr Ser Phe
500 505 510
Ile Arg Tyr Tyr Thr Arg Thr Leu Tyr Gln Phe Gln Phe Gln Glu Ala
515 520 525
Leu Cys Gln Ala Ala Lys His Glu Gly Pro Leu His Lys Cys Asp Ile
530 535 540
Ser Asn Ser Thr Glu Ala Gly Gln Lys Leu Phe Asn Met Leu Arg Leu
545 550 555 560
Gly Lys Ser Glu Pro Trp Thr Leu Ala Leu Glu Asn Val Val Gly Ala
565 570 575
Lys Asn Met Asn Val Arg Pro Leu Leu Asn Tyr Phe Glu Pro Leu Phe
580 585 590
Thr Trp Leu Lys Asp Gln Asn Lys Asn Ser Phe Val Gly Trp Ser Thr
595 600 605
Asp Trp Ser Pro Tyr Ala Asp Gln Ser Ile Lys Val Arg Ile Ser Leu
610 615 620
Lys Ser Ala Leu Gly Asp Lys Ala Tyr Glu Trp Asn Asp Asn Glu Met
625 630 635 640
Tyr Leu Phe Arg Ser Ser Val Ala Tyr Ala Met Arg Gln Tyr Phe Leu
645 650 655
Lys Val Lys Asn Gln Met Ile Leu Phe Gly Glu Glu Asp Val Arg Val
660 665 670
Ala Asn Leu Lys Pro Arg Ile Ser Phe Asn Phe Phe Val Thr Ala Pro
675 680 685
Lys Asn Val Ser Asp Ile Ile Pro Arg Thr Glu Val Glu Lys Ala Ile
690 695 700
Arg Met Ser Arg Ser Arg Ile Asn Asp Ala Phe Arg Leu Asn Asp Asn
705 710 715 720
Ser Leu Glu Phe Leu Gly Ile Gln Pro Thr Leu Gly Pro Pro Asn Gln
725 730 735
Pro Pro Val Ser Ile Trp Leu Ile Val Phe Gly Val Val Met Gly Val
740 745 750
Ile Val Val Gly Ile Val Ile Leu Ile Phe Thr Gly Ile Arg Asp Arg
755 760 765
Lys Lys Lys Asn Lys Ala Arg Ser Gly Glu Asn Pro Tyr Ala Ser Ile
770 775 780
Asp Ile Ser Lys Gly Glu Asn Asn Pro Gly Phe Gln Asn Thr Asp Asp
785 790 795 800
Val Gln Thr Ser Phe
805
<210> 2
<211> 2418
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 2
atgtcaagct cttcctggct ccttctcagc cttgttgctg taactgctgc tcagtccacc 60
attgaggaac aggccaagac atttttggac aagtttaacc acgaagccga agacctgttc 120
tatcaaagtt cacttgcttc ttggaattat aacaccaata ttactgaaga gaatgtccaa 180
aacatgaata atgctgggga caaatggtct gcctttttaa aggaacagtc cacacttgcc 240
caaatgtatc cactacaaga aattcagaat ctcacagtca agcttcagct gcaggctctt 300
cagcaaaatg ggtcttcagt gctctcagaa gacaagagca aacggttgaa cacaattcta 360
aatacaatga gcaccatcta cagtactgga aaagtttgta acccagataa tccacaagaa 420
tgcttattac ttgaaccagg tttgaatgaa ataatggcaa acagtttaga ctacaatgag 480
aggctctggg cttgggaaag ctggagatct gaggtcggca agcagctgag gccattatat 540
gaagagtatg tggtcttgaa aaatgagatg gcaagagcaa atcattatga ggactatggg 600
gattattgga gaggagacta tgaagtaaat ggggtagatg gctatgacta cagccgcggc 660
cagttgattg aagatgtgga acataccttt gaagagatta aaccattata tgaacatctt 720
catgcctatg tgagggcaaa gttgatgaat gcctatcctt cctatatcag tccaattgga 780
tgcctccctg ctcatttgct tggtgatatg tggggtagat tttggacaaa tctgtactct 840
ttgacagttc cctttggaca gaaaccaaac atagatgtta ctgatgcaat ggtggaccag 900
gcctgggatg cacagagaat attcaaggag gccgagaagt tctttgtatc tgttggtctt 960
cctaatatga ctcaaggatt ctgggaaaat tccatgctaa cggacccagg aaatgttcag 1020
aaagcagtct gccatcccac agcttgggac ctggggaagg gcgacttcag gatccttatg 1080
tgcacaaagg tgacaatgga cgacttcctg acagctcatc atgagatggg gcatatccag 1140
tatgatatgg catatgctgc acaacctttt ctgctaagaa atggagctaa tgaaggattc 1200
catgaagctg ttggggaaat catgtcactt tctgcagcca cacctaagca tttaaaatcc 1260
attggtcttc tgtcacccga ttttcaagaa gacaatgaaa cagaaataaa cttcctgctc 1320
aaacaagcac tcacgattgt tgggactctg ccatttactt acatgttaga gaagtggagg 1380
tggatggtct ttaaagggga aattcccaaa gaccagtgga tgaaaaagtg gtgggagatg 1440
aagcgagaga tagttggggt ggtggaacct gtgccccatg atgaaacata ctgtgacccc 1500
gcatctctgt tccatgtttc taatgattac tcattcattc gatattacac aaggaccctt 1560
taccaattcc agtttcaaga agcactttgt caagcagcta aacatgaagg ccctctgcac 1620
aaatgtgaca tctcaaactc tacagaagct ggacagaaac tgttcaatat gctgaggctt 1680
ggaaaatcag aaccctggac cctagcattg gaaaatgttg taggagcaaa gaacatgaat 1740
gtaaggccac tgctcaacta ctttgagccc ttatttacct ggctgaaaga ccagaacaag 1800
aattcttttg tgggatggag taccgactgg agtccatatg cagaccaaag catcaaagtg 1860
aggataagcc taaaatcagc tcttggagat aaagcatatg aatggaacga caatgaaatg 1920
tacctgttcc gatcatctgt tgcatatgct atgaggcagt actttttaaa agtaaaaaat 1980
cagatgattc tttttgggga ggaggatgtg cgagtggcta atttgaaacc aagaatctcc 2040
tttaatttct ttgtcactgc acctaaaaat gtgtctgata tcattcctag aactgaagtt 2100
gaaaaggcca tcaggatgtc ccggagccgt atcaatgatg ctttccgtct gaatgacaac 2160
agcctagagt ttctggggat acagccaaca cttggacctc ctaaccagcc ccctgtttcc 2220
atatggctga ttgtttttgg agttgtgatg ggagtgatag tggttggcat tgtcatcctg 2280
atcttcactg ggatcagaga tcggaagaag aaaaataaag caagaagtgg agaaaatcct 2340
tatgcctcca tcgatattag caaaggagaa aataatccag gattccaaaa cactgatgat 2400
gttcagacct ccttttag 2418
<210> 3
<211> 615
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 3
Met Ser Ser Ser Ser Trp Leu Leu Leu Ser Leu Val Ala Val Thr Ala
1 5 10 15
Ala Gln Ser Thr Ile Glu Glu Gln Ala Lys Thr Phe Leu Asp Lys Phe
20 25 30
Asn His Glu Ala Glu Asp Leu Phe Tyr Gln Ser Ser Leu Ala Ser Trp
35 40 45
Asn Tyr Asn Thr Asn Ile Thr Glu Glu Asn Val Gln Asn Met Asn Asn
50 55 60
Ala Gly Asp Lys Trp Ser Ala Phe Leu Lys Glu Gln Ser Thr Leu Ala
65 70 75 80
Gln Met Tyr Pro Leu Gln Glu Ile Gln Asn Leu Thr Val Lys Leu Gln
85 90 95
Leu Gln Ala Leu Gln Gln Asn Gly Ser Ser Val Leu Ser Glu Asp Lys
100 105 110
Ser Lys Arg Leu Asn Thr Ile Leu Asn Thr Met Ser Thr Ile Tyr Ser
115 120 125
Thr Gly Lys Val Cys Asn Pro Asp Asn Pro Gln Glu Cys Leu Leu Leu
130 135 140
Glu Pro Gly Leu Asn Glu Ile Met Ala Asn Ser Leu Asp Tyr Asn Glu
145 150 155 160
Arg Leu Trp Ala Trp Glu Ser Trp Arg Ser Glu Val Gly Lys Gln Leu
165 170 175
Arg Pro Leu Tyr Glu Glu Tyr Val Val Leu Lys Asn Glu Met Ala Arg
180 185 190
Ala Asn His Tyr Glu Asp Tyr Gly Asp Tyr Trp Arg Gly Asp Tyr Glu
195 200 205
Val Asn Gly Val Asp Gly Tyr Asp Tyr Ser Arg Gly Gln Leu Ile Glu
210 215 220
Asp Val Glu His Thr Phe Glu Glu Ile Lys Pro Leu Tyr Glu His Leu
225 230 235 240
His Ala Tyr Val Arg Ala Lys Leu Met Asn Ala Tyr Pro Ser Tyr Ile
245 250 255
Ser Pro Ile Gly Cys Leu Pro Ala His Leu Leu Gly Asp Met Trp Gly
260 265 270
Arg Phe Trp Thr Asn Leu Tyr Ser Leu Thr Val Pro Phe Gly Gln Lys
275 280 285
Pro Asn Ile Asp Val Thr Asp Ala Met Val Asp Gln Ala Trp Asp Ala
290 295 300
Gln Arg Ile Phe Lys Glu Ala Glu Lys Phe Phe Val Ser Val Gly Leu
305 310 315 320
Pro Asn Met Thr Gln Gly Phe Trp Glu Asn Ser Met Leu Thr Asp Pro
325 330 335
Gly Asn Val Gln Lys Ala Val Cys His Pro Thr Ala Trp Asp Leu Gly
340 345 350
Lys Gly Asp Phe Arg Ile Leu Met Cys Thr Lys Val Thr Met Asp Asp
355 360 365
Phe Leu Thr Ala His His Glu Met Gly His Ile Gln Tyr Asp Met Ala
370 375 380
Tyr Ala Ala Gln Pro Phe Leu Leu Arg Asn Gly Ala Asn Glu Gly Phe
385 390 395 400
His Glu Ala Val Gly Glu Ile Met Ser Leu Ser Ala Ala Thr Pro Lys
405 410 415
His Leu Lys Ser Ile Gly Leu Leu Ser Pro Asp Phe Gln Glu Asp Asn
420 425 430
Glu Thr Glu Ile Asn Phe Leu Leu Lys Gln Ala Leu Thr Ile Val Gly
435 440 445
Thr Leu Pro Phe Thr Tyr Met Leu Glu Lys Trp Arg Trp Met Val Phe
450 455 460
Lys Gly Glu Ile Pro Lys Asp Gln Trp Met Lys Lys Trp Trp Glu Met
465 470 475 480
Lys Arg Glu Ile Val Gly Val Val Glu Pro Val Pro His Asp Glu Thr
485 490 495
Tyr Cys Asp Pro Ala Ser Leu Phe His Val Ser Asn Asp Tyr Ser Phe
500 505 510
Ile Arg Tyr Tyr Thr Arg Thr Leu Tyr Gln Phe Gln Phe Gln Glu Ala
515 520 525
Leu Cys Gln Ala Ala Lys His Glu Gly Pro Leu His Lys Cys Asp Ile
530 535 540
Ser Asn Ser Thr Glu Ala Gly Gln Lys Leu Phe Asn Met Leu Arg Leu
545 550 555 560
Gly Lys Ser Glu Pro Trp Thr Leu Ala Leu Glu Asn Val Val Gly Ala
565 570 575
Lys Asn Met Asn Val Arg Pro Leu Leu Asn Tyr Phe Glu Pro Leu Phe
580 585 590
Thr Trp Leu Lys Asp Gln Asn Lys Asn Ser Phe Val Gly Trp Ser Thr
595 600 605
Asp Trp Ser Pro Tyr Ala Asp
610 615
<210> 4
<211> 1845
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 4
atgtcaagct cttcctggct ccttctcagc cttgttgctg taactgctgc tcagtccacc 60
attgaggaac aggccaagac atttttggac aagtttaacc acgaagccga agacctgttc 120
tatcaaagtt cacttgcttc ttggaattat aacaccaata ttactgaaga gaatgtccaa 180
aacatgaata atgctgggga caaatggtct gcctttttaa aggaacagtc cacacttgcc 240
caaatgtatc cactacaaga aattcagaat ctcacagtca agcttcagct gcaggctctt 300
cagcaaaatg ggtcttcagt gctctcagaa gacaagagca aacggttgaa cacaattcta 360
aatacaatga gcaccatcta cagtactgga aaagtttgta acccagataa tccacaagaa 420
tgcttattac ttgaaccagg tttgaatgaa ataatggcaa acagtttaga ctacaatgag 480
aggctctggg cttgggaaag ctggagatct gaggtcggca agcagctgag gccattatat 540
gaagagtatg tggtcttgaa aaatgagatg gcaagagcaa atcattatga ggactatggg 600
gattattgga gaggagacta tgaagtaaat ggggtagatg gctatgacta cagccgcggc 660
cagttgattg aagatgtgga acataccttt gaagagatta aaccattata tgaacatctt 720
catgcctatg tgagggcaaa gttgatgaat gcctatcctt cctatatcag tccaattgga 780
tgcctccctg ctcatttgct tggtgatatg tggggtagat tttggacaaa tctgtactct 840
ttgacagttc cctttggaca gaaaccaaac atagatgtta ctgatgcaat ggtggaccag 900
gcctgggatg cacagagaat attcaaggag gccgagaagt tctttgtatc tgttggtctt 960
cctaatatga ctcaaggatt ctgggaaaat tccatgctaa cggacccagg aaatgttcag 1020
aaagcagtct gccatcccac agcttgggac ctggggaagg gcgacttcag gatccttatg 1080
tgcacaaagg tgacaatgga cgacttcctg acagctcatc atgagatggg gcatatccag 1140
tatgatatgg catatgctgc acaacctttt ctgctaagaa atggagctaa tgaaggattc 1200
catgaagctg ttggggaaat catgtcactt tctgcagcca cacctaagca tttaaaatcc 1260
attggtcttc tgtcacccga ttttcaagaa gacaatgaaa cagaaataaa cttcctgctc 1320
aaacaagcac tcacgattgt tgggactctg ccatttactt acatgttaga gaagtggagg 1380
tggatggtct ttaaagggga aattcccaaa gaccagtgga tgaaaaagtg gtgggagatg 1440
aagcgagaga tagttggggt ggtggaacct gtgccccatg atgaaacata ctgtgacccc 1500
gcatctctgt tccatgtttc taatgattac tcattcattc gatattacac aaggaccctt 1560
taccaattcc agtttcaaga agcactttgt caagcagcta aacatgaagg ccctctgcac 1620
aaatgtgaca tctcaaactc tacagaagct ggacagaaac tgttcaatat gctgaggctt 1680
ggaaaatcag aaccctggac cctagcattg gaaaatgttg taggagcaaa gaacatgaat 1740
gtaaggccac tgctcaacta ctttgagccc ttatttacct ggctgaaaga ccagaacaag 1800
aattcttttg tgggatggag taccgactgg agtccatatg cagac 1845
<210> 5
<211> 232
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 5
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
1 5 10 15
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
20 25 30
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
35 40 45
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
50 55 60
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
65 70 75 80
Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
85 90 95
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
100 105 110
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
115 120 125
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr
130 135 140
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
145 150 155 160
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
165 170 175
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
180 185 190
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
195 200 205
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
210 215 220
Ser Leu Ser Leu Ser Pro Gly Lys
225 230
<210> 6
<211> 232
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 6
Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
1 5 10 15
Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
20 25 30
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
35 40 45
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
50 55 60
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
65 70 75 80
Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
85 90 95
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Ala Val Ser Asn Lys Ala
100 105 110
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
115 120 125
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr
130 135 140
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
145 150 155 160
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
165 170 175
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
180 185 190
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
195 200 205
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
210 215 220
Ser Leu Ser Leu Ser Pro Gly Lys
225 230
<210> 7
<211> 847
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 7
Met Ser Ser Ser Ser Trp Leu Leu Leu Ser Leu Val Ala Val Thr Ala
1 5 10 15
Ala Gln Ser Thr Ile Glu Glu Gln Ala Lys Thr Phe Leu Asp Lys Phe
20 25 30
Asn His Glu Ala Glu Asp Leu Phe Tyr Gln Ser Ser Leu Ala Ser Trp
35 40 45
Asn Tyr Asn Thr Asn Ile Thr Glu Glu Asn Val Gln Asn Met Asn Asn
50 55 60
Ala Gly Asp Lys Trp Ser Ala Phe Leu Lys Glu Gln Ser Thr Leu Ala
65 70 75 80
Gln Met Tyr Pro Leu Gln Glu Ile Gln Asn Leu Thr Val Lys Leu Gln
85 90 95
Leu Gln Ala Leu Gln Gln Asn Gly Ser Ser Val Leu Ser Glu Asp Lys
100 105 110
Ser Lys Arg Leu Asn Thr Ile Leu Asn Thr Met Ser Thr Ile Tyr Ser
115 120 125
Thr Gly Lys Val Cys Asn Pro Asp Asn Pro Gln Glu Cys Leu Leu Leu
130 135 140
Glu Pro Gly Leu Asn Glu Ile Met Ala Asn Ser Leu Asp Tyr Asn Glu
145 150 155 160
Arg Leu Trp Ala Trp Glu Ser Trp Arg Ser Glu Val Gly Lys Gln Leu
165 170 175
Arg Pro Leu Tyr Glu Glu Tyr Val Val Leu Lys Asn Glu Met Ala Arg
180 185 190
Ala Asn His Tyr Glu Asp Tyr Gly Asp Tyr Trp Arg Gly Asp Tyr Glu
195 200 205
Val Asn Gly Val Asp Gly Tyr Asp Tyr Ser Arg Gly Gln Leu Ile Glu
210 215 220
Asp Val Glu His Thr Phe Glu Glu Ile Lys Pro Leu Tyr Glu His Leu
225 230 235 240
His Ala Tyr Val Arg Ala Lys Leu Met Asn Ala Tyr Pro Ser Tyr Ile
245 250 255
Ser Pro Ile Gly Cys Leu Pro Ala His Leu Leu Gly Asp Met Trp Gly
260 265 270
Arg Phe Trp Thr Asn Leu Tyr Ser Leu Thr Val Pro Phe Gly Gln Lys
275 280 285
Pro Asn Ile Asp Val Thr Asp Ala Met Val Asp Gln Ala Trp Asp Ala
290 295 300
Gln Arg Ile Phe Lys Glu Ala Glu Lys Phe Phe Val Ser Val Gly Leu
305 310 315 320
Pro Asn Met Thr Gln Gly Phe Trp Glu Asn Ser Met Leu Thr Asp Pro
325 330 335
Gly Asn Val Gln Lys Ala Val Cys His Pro Thr Ala Trp Asp Leu Gly
340 345 350
Lys Gly Asp Phe Arg Ile Leu Met Cys Thr Lys Val Thr Met Asp Asp
355 360 365
Phe Leu Thr Ala His His Glu Met Gly His Ile Gln Tyr Asp Met Ala
370 375 380
Tyr Ala Ala Gln Pro Phe Leu Leu Arg Asn Gly Ala Asn Glu Gly Phe
385 390 395 400
His Glu Ala Val Gly Glu Ile Met Ser Leu Ser Ala Ala Thr Pro Lys
405 410 415
His Leu Lys Ser Ile Gly Leu Leu Ser Pro Asp Phe Gln Glu Asp Asn
420 425 430
Glu Thr Glu Ile Asn Phe Leu Leu Lys Gln Ala Leu Thr Ile Val Gly
435 440 445
Thr Leu Pro Phe Thr Tyr Met Leu Glu Lys Trp Arg Trp Met Val Phe
450 455 460
Lys Gly Glu Ile Pro Lys Asp Gln Trp Met Lys Lys Trp Trp Glu Met
465 470 475 480
Lys Arg Glu Ile Val Gly Val Val Glu Pro Val Pro His Asp Glu Thr
485 490 495
Tyr Cys Asp Pro Ala Ser Leu Phe His Val Ser Asn Asp Tyr Ser Phe
500 505 510
Ile Arg Tyr Tyr Thr Arg Thr Leu Tyr Gln Phe Gln Phe Gln Glu Ala
515 520 525
Leu Cys Gln Ala Ala Lys His Glu Gly Pro Leu His Lys Cys Asp Ile
530 535 540
Ser Asn Ser Thr Glu Ala Gly Gln Lys Leu Phe Asn Met Leu Arg Leu
545 550 555 560
Gly Lys Ser Glu Pro Trp Thr Leu Ala Leu Glu Asn Val Val Gly Ala
565 570 575
Lys Asn Met Asn Val Arg Pro Leu Leu Asn Tyr Phe Glu Pro Leu Phe
580 585 590
Thr Trp Leu Lys Asp Gln Asn Lys Asn Ser Phe Val Gly Trp Ser Thr
595 600 605
Asp Trp Ser Pro Tyr Ala Asp Glu Pro Lys Ser Ser Asp Lys Thr His
610 615 620
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val
625 630 635 640
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
645 650 655
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
660 665 670
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
675 680 685
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
690 695 700
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
705 710 715 720
Cys Ala Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
725 730 735
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
740 745 750
Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu
755 760 765
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
770 775 780
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
785 790 795 800
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
805 810 815
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
820 825 830
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
835 840 845
<210> 8
<211> 2544
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 8
atgtcaagct cttcctggct ccttctcagc cttgttgctg taactgctgc tcagtccacc 60
attgaggaac aggccaagac atttttggac aagtttaacc acgaagccga agacctgttc 120
tatcaaagtt cacttgcttc ttggaattat aacaccaata ttactgaaga gaatgtccaa 180
aacatgaata atgctgggga caaatggtct gcctttttaa aggaacagtc cacacttgcc 240
caaatgtatc cactacaaga aattcagaat ctcacagtca agcttcagct gcaggctctt 300
cagcaaaatg ggtcttcagt gctctcagaa gacaagagca aacggttgaa cacaattcta 360
aatacaatga gcaccatcta cagtactgga aaagtttgta acccagataa tccacaagaa 420
tgcttattac ttgaaccagg tttgaatgaa ataatggcaa acagtttaga ctacaatgag 480
aggctctggg cttgggaaag ctggagatct gaggtcggca agcagctgag gccattatat 540
gaagagtatg tggtcttgaa aaatgagatg gcaagagcaa atcattatga ggactatggg 600
gattattgga gaggagacta tgaagtaaat ggggtagatg gctatgacta cagccgcggc 660
cagttgattg aagatgtgga acataccttt gaagagatta aaccattata tgaacatctt 720
catgcctatg tgagggcaaa gttgatgaat gcctatcctt cctatatcag tccaattgga 780
tgcctccctg ctcatttgct tggtgatatg tggggtagat tttggacaaa tctgtactct 840
ttgacagttc cctttggaca gaaaccaaac atagatgtta ctgatgcaat ggtggaccag 900
gcctgggatg cacagagaat attcaaggag gccgagaagt tctttgtatc tgttggtctt 960
cctaatatga ctcaaggatt ctgggaaaat tccatgctaa cggacccagg aaatgttcag 1020
aaagcagtct gccatcccac agcttgggac ctggggaagg gcgacttcag gatccttatg 1080
tgcacaaagg tgacaatgga cgacttcctg acagctcatc atgagatggg gcatatccag 1140
tatgatatgg catatgctgc acaacctttt ctgctaagaa atggagctaa tgaaggattc 1200
catgaagctg ttggggaaat catgtcactt tctgcagcca cacctaagca tttaaaatcc 1260
attggtcttc tgtcacccga ttttcaagaa gacaatgaaa cagaaataaa cttcctgctc 1320
aaacaagcac tcacgattgt tgggactctg ccatttactt acatgttaga gaagtggagg 1380
tggatggtct ttaaagggga aattcccaaa gaccagtgga tgaaaaagtg gtgggagatg 1440
aagcgagaga tagttggggt ggtggaacct gtgccccatg atgaaacata ctgtgacccc 1500
gcatctctgt tccatgtttc taatgattac tcattcattc gatattacac aaggaccctt 1560
taccaattcc agtttcaaga agcactttgt caagcagcta aacatgaagg ccctctgcac 1620
aaatgtgaca tctcaaactc tacagaagct ggacagaaac tgttcaatat gctgaggctt 1680
ggaaaatcag aaccctggac cctagcattg gaaaatgttg taggagcaaa gaacatgaat 1740
gtaaggccac tgctcaacta ctttgagccc ttatttacct ggctgaaaga ccagaacaag 1800
aattcttttg tgggatggag taccgactgg agtccatatg cagacgagcc caaatcttcc 1860
gacaaaactc acacatgccc accgtgccca gcacctgaag ccgcgggggg accgtcagtc 1920
ttcctcttcc ccccaaaacc caaggacacc ctcatgatct cccggacccc tgaggtcaca 1980
tgcgtggtgg tggacgtgag ccacgaagac cctgaggtca agttcaactg gtacgtggac 2040
ggcgtggagg tgcataatgc caagacaaag ccgcgggagg agcagtacaa cagcacgtac 2100
cgtgtggtca gcgtcctcac cgtcctgcac caggactggc tgaatggcaa ggagtacaag 2160
tgcgcggtct ccaacaaagc cctcccagcc cccatcgaga aaaccatctc caaagccaaa 2220
gggcagcccc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgaccaag 2280
aaccaggtca gcctgacctg cctggtcaaa ggcttctatc ccagcgacat cgccgtggag 2340
tgggagagca atgggcagcc ggagaacaac tacaagacca cgcctcccgt gctggactcc 2400
gacggctcct tcttcctcta tagcaagctc accgtggaca agagcaggtg gcagcagggg 2460
aacgtcttct catgctccgt gatgcatgag gctctgcaca accactacac gcagaagagc 2520
ctctccctgt ctccgggtaa atag 2544
<210> 9
<211> 844
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 9
Met Ser Ser Ser Ser Trp Leu Leu Leu Ser Leu Val Ala Val Thr Ala
1 5 10 15
Ala Gln Ser Thr Ile Glu Glu Gln Ala Lys Thr Phe Leu Asp Lys Phe
20 25 30
Asn His Glu Ala Glu Asp Leu Phe Tyr Gln Ser Ser Leu Ala Ser Trp
35 40 45
Asn Tyr Asn Thr Asn Ile Thr Glu Glu Asn Val Gln Asn Met Asn Asn
50 55 60
Ala Gly Asp Lys Trp Ser Ala Phe Leu Lys Glu Gln Ser Thr Leu Ala
65 70 75 80
Gln Met Tyr Pro Leu Gln Glu Ile Gln Asn Leu Thr Val Lys Leu Gln
85 90 95
Leu Gln Ala Leu Gln Gln Asn Gly Ser Ser Val Leu Ser Glu Asp Lys
100 105 110
Ser Lys Arg Leu Asn Thr Ile Leu Asn Thr Met Ser Thr Ile Tyr Ser
115 120 125
Thr Gly Lys Val Cys Asn Pro Asp Asn Pro Gln Glu Cys Leu Leu Leu
130 135 140
Glu Pro Gly Leu Asn Glu Ile Met Ala Asn Ser Leu Asp Tyr Asn Glu
145 150 155 160
Arg Leu Trp Ala Trp Glu Ser Trp Arg Ser Glu Val Gly Lys Gln Leu
165 170 175
Arg Pro Leu Tyr Glu Glu Tyr Val Val Leu Lys Asn Glu Met Ala Arg
180 185 190
Ala Asn His Tyr Glu Asp Tyr Gly Asp Tyr Trp Arg Gly Asp Tyr Glu
195 200 205
Val Asn Gly Val Asp Gly Tyr Asp Tyr Ser Arg Gly Gln Leu Ile Glu
210 215 220
Asp Val Glu His Thr Phe Glu Glu Ile Lys Pro Leu Tyr Glu His Leu
225 230 235 240
His Ala Tyr Val Arg Ala Lys Leu Met Asn Ala Tyr Pro Ser Tyr Ile
245 250 255
Ser Pro Ile Gly Cys Leu Pro Ala His Leu Leu Gly Asp Met Trp Gly
260 265 270
Arg Phe Trp Thr Asn Leu Tyr Ser Leu Thr Val Pro Phe Gly Gln Lys
275 280 285
Pro Asn Ile Asp Val Thr Asp Ala Met Val Asp Gln Ala Trp Asp Ala
290 295 300
Gln Arg Ile Phe Lys Glu Ala Glu Lys Phe Phe Val Ser Val Gly Leu
305 310 315 320
Pro Asn Met Thr Gln Gly Phe Trp Glu Asn Ser Met Leu Thr Asp Pro
325 330 335
Gly Asn Val Gln Lys Ala Val Cys His Pro Thr Ala Trp Asp Leu Gly
340 345 350
Lys Gly Asp Phe Arg Ile Leu Met Cys Thr Lys Val Thr Met Asp Asp
355 360 365
Phe Leu Thr Ala His His Glu Met Gly His Ile Gln Tyr Asp Met Ala
370 375 380
Tyr Ala Ala Gln Pro Phe Leu Leu Arg Asn Gly Ala Asn Glu Gly Phe
385 390 395 400
His Glu Ala Val Gly Glu Ile Met Ser Leu Ser Ala Ala Thr Pro Lys
405 410 415
His Leu Lys Ser Ile Gly Leu Leu Ser Pro Asp Phe Gln Glu Asp Asn
420 425 430
Glu Thr Glu Ile Asn Phe Leu Leu Lys Gln Ala Leu Thr Ile Val Gly
435 440 445
Thr Leu Pro Phe Thr Tyr Met Leu Glu Lys Trp Arg Trp Met Val Phe
450 455 460
Lys Gly Glu Ile Pro Lys Asp Gln Trp Met Lys Lys Trp Trp Glu Met
465 470 475 480
Lys Arg Glu Ile Val Gly Val Val Glu Pro Val Pro His Asp Glu Thr
485 490 495
Tyr Cys Asp Pro Ala Ser Leu Phe His Val Ser Asn Asp Tyr Ser Phe
500 505 510
Ile Arg Tyr Tyr Thr Arg Thr Leu Tyr Gln Phe Gln Phe Gln Glu Ala
515 520 525
Leu Cys Gln Ala Ala Lys His Glu Gly Pro Leu His Lys Cys Asp Ile
530 535 540
Ser Asn Ser Thr Glu Ala Gly Gln Lys Leu Phe Asn Met Leu Arg Leu
545 550 555 560
Gly Lys Ser Glu Pro Trp Thr Leu Ala Leu Glu Asn Val Val Gly Ala
565 570 575
Lys Asn Met Asn Val Arg Pro Leu Leu Asn Tyr Phe Glu Pro Leu Phe
580 585 590
Thr Trp Leu Lys Asp Gln Asn Lys Asn Ser Phe Val Gly Trp Ser Thr
595 600 605
Asp Trp Ser Pro Tyr Ala Asp Glu Ser Lys Tyr Gly Pro Pro Cys Pro
610 615 620
Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe
625 630 635 640
Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
645 650 655
Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe
660 665 670
Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro
675 680 685
Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr
690 695 700
Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val
705 710 715 720
Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala
725 730 735
Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln
740 745 750
Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly
755 760 765
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro
770 775 780
Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser
785 790 795 800
Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu
805 810 815
Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His
820 825 830
Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys
835 840
<210> 10
<211> 2535
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 10
atgtcaagct cttcctggct ccttctcagc cttgttgctg taactgctgc tcagtccacc 60
attgaggaac aggccaagac atttttggac aagtttaacc acgaagccga agacctgttc 120
tatcaaagtt cacttgcttc ttggaattat aacaccaata ttactgaaga gaatgtccaa 180
aacatgaata atgctgggga caaatggtct gcctttttaa aggaacagtc cacacttgcc 240
caaatgtatc cactacaaga aattcagaat ctcacagtca agcttcagct gcaggctctt 300
cagcaaaatg ggtcttcagt gctctcagaa gacaagagca aacggttgaa cacaattcta 360
aatacaatga gcaccatcta cagtactgga aaagtttgta acccagataa tccacaagaa 420
tgcttattac ttgaaccagg tttgaatgaa ataatggcaa acagtttaga ctacaatgag 480
aggctctggg cttgggaaag ctggagatct gaggtcggca agcagctgag gccattatat 540
gaagagtatg tggtcttgaa aaatgagatg gcaagagcaa atcattatga ggactatggg 600
gattattgga gaggagacta tgaagtaaat ggggtagatg gctatgacta cagccgcggc 660
cagttgattg aagatgtgga acataccttt gaagagatta aaccattata tgaacatctt 720
catgcctatg tgagggcaaa gttgatgaat gcctatcctt cctatatcag tccaattgga 780
tgcctccctg ctcatttgct tggtgatatg tggggtagat tttggacaaa tctgtactct 840
ttgacagttc cctttggaca gaaaccaaac atagatgtta ctgatgcaat ggtggaccag 900
gcctgggatg cacagagaat attcaaggag gccgagaagt tctttgtatc tgttggtctt 960
cctaatatga ctcaaggatt ctgggaaaat tccatgctaa cggacccagg aaatgttcag 1020
aaagcagtct gccatcccac agcttgggac ctggggaagg gcgacttcag gatccttatg 1080
tgcacaaagg tgacaatgga cgacttcctg acagctcatc atgagatggg gcatatccag 1140
tatgatatgg catatgctgc acaacctttt ctgctaagaa atggagctaa tgaaggattc 1200
catgaagctg ttggggaaat catgtcactt tctgcagcca cacctaagca tttaaaatcc 1260
attggtcttc tgtcacccga ttttcaagaa gacaatgaaa cagaaataaa cttcctgctc 1320
aaacaagcac tcacgattgt tgggactctg ccatttactt acatgttaga gaagtggagg 1380
tggatggtct ttaaagggga aattcccaaa gaccagtgga tgaaaaagtg gtgggagatg 1440
aagcgagaga tagttggggt ggtggaacct gtgccccatg atgaaacata ctgtgacccc 1500
gcatctctgt tccatgtttc taatgattac tcattcattc gatattacac aaggaccctt 1560
taccaattcc agtttcaaga agcactttgt caagcagcta aacatgaagg ccctctgcac 1620
aaatgtgaca tctcaaactc tacagaagct ggacagaaac tgttcaatat gctgaggctt 1680
ggaaaatcag aaccctggac cctagcattg gaaaatgttg taggagcaaa gaacatgaat 1740
gtaaggccac tgctcaacta ctttgagccc ttatttacct ggctgaaaga ccagaacaag 1800
aattcttttg tgggatggag taccgactgg agtccatatg cagacgagtc caaatatggt 1860
cccccgtgcc caccatgccc agcacctgag ttcctggggg gaccatcagt cttcctgttc 1920
cccccaaaac ccaaggacac tctcatgatc tcccggaccc ctgaggtcac gtgcgtggtg 1980
gtggacgtga gccaggaaga ccccgaggtc cagttcaact ggtacgtgga tggcgtggag 2040
gtgcataatg ccaagacaaa gccgcgggag gagcagttca acagcacgta ccgtgtggtc 2100
agcgtcctca ccgtcctcca ccaggactgg ctgaacggca aggagtacaa gtgcaaggtc 2160
tccaacaaag gcctcccgtc ctccatcgag aaaaccatct ccaaagccaa agggcagccc 2220
cgagagccac aggtgtacac cctgccccca tcccaggagg agatgaccaa gaaccaggtc 2280
agcctgacct gcctggtcaa aggcttctac cccagcgaca tcgccgtgga gtgggagagc 2340
aatgggcagc cggagaacaa ctacaagacc acgcctcccg tgctggactc cgacggctcc 2400
ttcttcctct acagcaggct aaccgtggac aagagcaggt ggcaggaggg gaatgtcttc 2460
tcatgctccg tgatgcatga ggctctgcac aaccactaca cgcagaagag cctctccctg 2520
tctctgggta aatga 2535
<210> 11
<211> 875
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 11
Met Ser Ser Ser Ser Trp Leu Leu Leu Ser Leu Val Ala Val Thr Ala
1 5 10 15
Ala Gln Ser Thr Ile Glu Glu Gln Ala Lys Thr Phe Leu Asp Lys Phe
20 25 30
Asn His Glu Ala Glu Asp Leu Phe Tyr Gln Ser Ser Leu Ala Ser Trp
35 40 45
Asn Tyr Asn Thr Asn Ile Thr Glu Glu Asn Val Gln Asn Met Asn Asn
50 55 60
Ala Gly Asp Lys Trp Ser Ala Phe Leu Lys Glu Gln Ser Thr Leu Ala
65 70 75 80
Gln Met Tyr Pro Leu Gln Glu Ile Gln Asn Leu Thr Val Lys Leu Gln
85 90 95
Leu Gln Ala Leu Gln Gln Asn Gly Ser Ser Val Leu Ser Glu Asp Lys
100 105 110
Ser Lys Arg Leu Asn Thr Ile Leu Asn Thr Met Ser Thr Ile Tyr Ser
115 120 125
Thr Gly Lys Val Cys Asn Pro Asp Asn Pro Gln Glu Cys Leu Leu Leu
130 135 140
Glu Pro Gly Leu Asn Glu Ile Met Ala Asn Ser Leu Asp Tyr Asn Glu
145 150 155 160
Arg Leu Trp Ala Trp Glu Ser Trp Arg Ser Glu Val Gly Lys Gln Leu
165 170 175
Arg Pro Leu Tyr Glu Glu Tyr Val Val Leu Lys Asn Glu Met Ala Arg
180 185 190
Ala Asn His Tyr Glu Asp Tyr Gly Asp Tyr Trp Arg Gly Asp Tyr Glu
195 200 205
Val Asn Gly Val Asp Gly Tyr Asp Tyr Ser Arg Gly Gln Leu Ile Glu
210 215 220
Asp Val Glu His Thr Phe Glu Glu Ile Lys Pro Leu Tyr Glu His Leu
225 230 235 240
His Ala Tyr Val Arg Ala Lys Leu Met Asn Ala Tyr Pro Ser Tyr Ile
245 250 255
Ser Pro Ile Gly Cys Leu Pro Ala His Leu Leu Gly Asp Met Trp Gly
260 265 270
Arg Phe Trp Thr Asn Leu Tyr Ser Leu Thr Val Pro Phe Gly Gln Lys
275 280 285
Pro Asn Ile Asp Val Thr Asp Ala Met Val Asp Gln Ala Trp Asp Ala
290 295 300
Gln Arg Ile Phe Lys Glu Ala Glu Lys Phe Phe Val Ser Val Gly Leu
305 310 315 320
Pro Asn Met Thr Gln Gly Phe Trp Glu Asn Ser Met Leu Thr Asp Pro
325 330 335
Gly Asn Val Gln Lys Ala Val Cys His Pro Thr Ala Trp Asp Leu Gly
340 345 350
Lys Gly Asp Phe Arg Ile Leu Met Cys Thr Lys Val Thr Met Asp Asp
355 360 365
Phe Leu Thr Ala His His Glu Met Gly His Ile Gln Tyr Asp Met Ala
370 375 380
Tyr Ala Ala Gln Pro Phe Leu Leu Arg Asn Gly Ala Asn Glu Gly Phe
385 390 395 400
His Glu Ala Val Gly Glu Ile Met Ser Leu Ser Ala Ala Thr Pro Lys
405 410 415
His Leu Lys Ser Ile Gly Leu Leu Ser Pro Asp Phe Gln Glu Asp Asn
420 425 430
Glu Thr Glu Ile Asn Phe Leu Leu Lys Gln Ala Leu Thr Ile Val Gly
435 440 445
Thr Leu Pro Phe Thr Tyr Met Leu Glu Lys Trp Arg Trp Met Val Phe
450 455 460
Lys Gly Glu Ile Pro Lys Asp Gln Trp Met Lys Lys Trp Trp Glu Met
465 470 475 480
Lys Arg Glu Ile Val Gly Val Val Glu Pro Val Pro His Asp Glu Thr
485 490 495
Tyr Cys Asp Pro Ala Ser Leu Phe His Val Ser Asn Asp Tyr Ser Phe
500 505 510
Ile Arg Tyr Tyr Thr Arg Thr Leu Tyr Gln Phe Gln Phe Gln Glu Ala
515 520 525
Leu Cys Gln Ala Ala Lys His Glu Gly Pro Leu His Lys Cys Asp Ile
530 535 540
Ser Asn Ser Thr Glu Ala Gly Gln Lys Leu Phe Asn Met Leu Arg Leu
545 550 555 560
Gly Lys Ser Glu Pro Trp Thr Leu Ala Leu Glu Asn Val Val Gly Ala
565 570 575
Lys Asn Met Asn Val Arg Pro Leu Leu Asn Tyr Phe Glu Pro Leu Phe
580 585 590
Thr Trp Leu Lys Asp Gln Asn Lys Asn Ser Phe Val Gly Trp Ser Thr
595 600 605
Asp Trp Ser Pro Tyr Ala Asp Ser Gln Asp Val Thr Val Pro Cys Pro
610 615 620
Val Pro Ser Thr Pro Pro Thr Pro Ser Pro Ser Thr Pro Pro Thr Pro
625 630 635 640
Ser Pro Ser Cys Cys His Pro Arg Leu Ser Leu His Arg Pro Ala Leu
645 650 655
Glu Asp Leu Leu Leu Gly Ser Glu Ala Asn Leu Thr Cys Thr Leu Thr
660 665 670
Gly Leu Arg Asp Ala Ser Gly Val Thr Phe Thr Trp Thr Pro Ser Ser
675 680 685
Gly Lys Ser Ala Val Gln Gly Pro Pro Glu Arg Asp Leu Cys Gly Cys
690 695 700
Tyr Ser Val Ser Ser Val Leu Pro Gly Cys Ala Glu Pro Trp Asn His
705 710 715 720
Gly Lys Thr Phe Thr Cys Thr Ala Ala Tyr Pro Glu Ser Lys Thr Pro
725 730 735
Leu Thr Ala Thr Leu Ser Lys Ser Gly Asn Thr Phe Arg Pro Glu Val
740 745 750
His Leu Leu Pro Pro Pro Ser Glu Glu Leu Ala Leu Asn Glu Leu Val
755 760 765
Thr Leu Thr Cys Leu Ala Arg Gly Phe Ser Pro Lys Asp Val Leu Val
770 775 780
Arg Trp Leu Gln Gly Ser Gln Glu Leu Pro Arg Glu Lys Tyr Leu Thr
785 790 795 800
Trp Ala Ser Arg Gln Glu Pro Ser Gln Gly Thr Thr Thr Phe Ala Val
805 810 815
Thr Ser Ile Leu Arg Val Ala Ala Glu Asp Trp Lys Lys Gly Asp Thr
820 825 830
Phe Ser Cys Met Val Gly His Glu Ala Leu Pro Leu Ala Phe Thr Gln
835 840 845
Lys Thr Ile Asp Arg Leu Ala Gly Lys Pro Thr His Val Asn Val Ser
850 855 860
Val Val Met Ala Glu Val Asp Gly Thr Cys Tyr
865 870 875
<210> 12
<211> 2628
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 12
atgtcaagct cttcctggct ccttctcagc cttgttgctg taactgctgc tcagtccacc 60
attgaggaac aggccaagac atttttggac aagtttaacc acgaagccga agacctgttc 120
tatcaaagtt cacttgcttc ttggaattat aacaccaata ttactgaaga gaatgtccaa 180
aacatgaata atgctgggga caaatggtct gcctttttaa aggaacagtc cacacttgcc 240
caaatgtatc cactacaaga aattcagaat ctcacagtca agcttcagct gcaggctctt 300
cagcaaaatg ggtcttcagt gctctcagaa gacaagagca aacggttgaa cacaattcta 360
aatacaatga gcaccatcta cagtactgga aaagtttgta acccagataa tccacaagaa 420
tgcttattac ttgaaccagg tttgaatgaa ataatggcaa acagtttaga ctacaatgag 480
aggctctggg cttgggaaag ctggagatct gaggtcggca agcagctgag gccattatat 540
gaagagtatg tggtcttgaa aaatgagatg gcaagagcaa atcattatga ggactatggg 600
gattattgga gaggagacta tgaagtaaat ggggtagatg gctatgacta cagccgcggc 660
cagttgattg aagatgtgga acataccttt gaagagatta aaccattata tgaacatctt 720
catgcctatg tgagggcaaa gttgatgaat gcctatcctt cctatatcag tccaattgga 780
tgcctccctg ctcatttgct tggtgatatg tggggtagat tttggacaaa tctgtactct 840
ttgacagttc cctttggaca gaaaccaaac atagatgtta ctgatgcaat ggtggaccag 900
gcctgggatg cacagagaat attcaaggag gccgagaagt tctttgtatc tgttggtctt 960
cctaatatga ctcaaggatt ctgggaaaat tccatgctaa cggacccagg aaatgttcag 1020
aaagcagtct gccatcccac agcttgggac ctggggaagg gcgacttcag gatccttatg 1080
tgcacaaagg tgacaatgga cgacttcctg acagctcatc atgagatggg gcatatccag 1140
tatgatatgg catatgctgc acaacctttt ctgctaagaa atggagctaa tgaaggattc 1200
catgaagctg ttggggaaat catgtcactt tctgcagcca cacctaagca tttaaaatcc 1260
attggtcttc tgtcacccga ttttcaagaa gacaatgaaa cagaaataaa cttcctgctc 1320
aaacaagcac tcacgattgt tgggactctg ccatttactt acatgttaga gaagtggagg 1380
tggatggtct ttaaagggga aattcccaaa gaccagtgga tgaaaaagtg gtgggagatg 1440
aagcgagaga tagttggggt ggtggaacct gtgccccatg atgaaacata ctgtgacccc 1500
gcatctctgt tccatgtttc taatgattac tcattcattc gatattacac aaggaccctt 1560
taccaattcc agtttcaaga agcactttgt caagcagcta aacatgaagg ccctctgcac 1620
aaatgtgaca tctcaaactc tacagaagct ggacagaaac tgttcaatat gctgaggctt 1680
ggaaaatcag aaccctggac cctagcattg gaaaatgttg taggagcaaa gaacatgaat 1740
gtaaggccac tgctcaacta ctttgagccc ttatttacct ggctgaaaga ccagaacaag 1800
aattcttttg tgggatggag taccgactgg agtccatatg cagacagcca ggatgtgact 1860
gtgccctgcc cagttccctc aactccacct accccatctc cctcaactcc acctacccca 1920
tctccctcat gctgccaccc ccgactgtca ctgcaccgac cggccctcga ggacctgctc 1980
ttaggttcag aagcgaacct cacgtgcaca ctgaccggcc tgagagatgc ctcaggtgtc 2040
accttcacct ggacgccctc aagtgggaag agcgctgttc aaggaccacc tgagcgtgac 2100
ctctgtggct gctacagcgt gtccagtgtc ctgccgggct gtgccgagcc atggaaccat 2160
gggaagacct tcacttgcac tgctgcctac cccgagtcca agaccccgct aaccgccacc 2220
ctctcaaaat ccggaaacac attccggccc gaggtccacc tgctgccgcc gccgtcggag 2280
gagctggccc tgaacgagct ggtgacgctg acgtgcctgg cacgcggctt cagccccaag 2340
gacgtgctgg ttcgctggct gcaggggtca caggagctgc cccgcgagaa gtacctgact 2400
tgggcatccc ggcaggagcc cagccagggc accaccacct tcgctgtgac cagcatactg 2460
cgcgtggcag ccgaggactg gaagaagggg gacaccttct cctgcatggt gggccacgag 2520
gccctgccgc tggccttcac acagaagacc atcgaccgct tggcgggtaa acccacccat 2580
gtcaatgtgt ctgttgtcat ggcggaggtg gacggcacct gctactga 2628
<210> 13
<211> 862
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 13
Met Ser Ser Ser Ser Trp Leu Leu Leu Ser Leu Val Ala Val Thr Ala
1 5 10 15
Ala Gln Ser Thr Ile Glu Glu Gln Ala Lys Thr Phe Leu Asp Lys Phe
20 25 30
Asn His Glu Ala Glu Asp Leu Phe Tyr Gln Ser Ser Leu Ala Ser Trp
35 40 45
Asn Tyr Asn Thr Asn Ile Thr Glu Glu Asn Val Gln Asn Met Asn Asn
50 55 60
Ala Gly Asp Lys Trp Ser Ala Phe Leu Lys Glu Gln Ser Thr Leu Ala
65 70 75 80
Gln Met Tyr Pro Leu Gln Glu Ile Gln Asn Leu Thr Val Lys Leu Gln
85 90 95
Leu Gln Ala Leu Gln Gln Asn Gly Ser Ser Val Leu Ser Glu Asp Lys
100 105 110
Ser Lys Arg Leu Asn Thr Ile Leu Asn Thr Met Ser Thr Ile Tyr Ser
115 120 125
Thr Gly Lys Val Cys Asn Pro Asp Asn Pro Gln Glu Cys Leu Leu Leu
130 135 140
Glu Pro Gly Leu Asn Glu Ile Met Ala Asn Ser Leu Asp Tyr Asn Glu
145 150 155 160
Arg Leu Trp Ala Trp Glu Ser Trp Arg Ser Glu Val Gly Lys Gln Leu
165 170 175
Arg Pro Leu Tyr Glu Glu Tyr Val Val Leu Lys Asn Glu Met Ala Arg
180 185 190
Ala Asn His Tyr Glu Asp Tyr Gly Asp Tyr Trp Arg Gly Asp Tyr Glu
195 200 205
Val Asn Gly Val Asp Gly Tyr Asp Tyr Ser Arg Gly Gln Leu Ile Glu
210 215 220
Asp Val Glu His Thr Phe Glu Glu Ile Lys Pro Leu Tyr Glu His Leu
225 230 235 240
His Ala Tyr Val Arg Ala Lys Leu Met Asn Ala Tyr Pro Ser Tyr Ile
245 250 255
Ser Pro Ile Gly Cys Leu Pro Ala His Leu Leu Gly Asp Met Trp Gly
260 265 270
Arg Phe Trp Thr Asn Leu Tyr Ser Leu Thr Val Pro Phe Gly Gln Lys
275 280 285
Pro Asn Ile Asp Val Thr Asp Ala Met Val Asp Gln Ala Trp Asp Ala
290 295 300
Gln Arg Ile Phe Lys Glu Ala Glu Lys Phe Phe Val Ser Val Gly Leu
305 310 315 320
Pro Asn Met Thr Gln Gly Phe Trp Glu Asn Ser Met Leu Thr Asp Pro
325 330 335
Gly Asn Val Gln Lys Ala Val Cys His Pro Thr Ala Trp Asp Leu Gly
340 345 350
Lys Gly Asp Phe Arg Ile Leu Met Cys Thr Lys Val Thr Met Asp Asp
355 360 365
Phe Leu Thr Ala His His Glu Met Gly His Ile Gln Tyr Asp Met Ala
370 375 380
Tyr Ala Ala Gln Pro Phe Leu Leu Arg Asn Gly Ala Asn Glu Gly Phe
385 390 395 400
His Glu Ala Val Gly Glu Ile Met Ser Leu Ser Ala Ala Thr Pro Lys
405 410 415
His Leu Lys Ser Ile Gly Leu Leu Ser Pro Asp Phe Gln Glu Asp Asn
420 425 430
Glu Thr Glu Ile Asn Phe Leu Leu Lys Gln Ala Leu Thr Ile Val Gly
435 440 445
Thr Leu Pro Phe Thr Tyr Met Leu Glu Lys Trp Arg Trp Met Val Phe
450 455 460
Lys Gly Glu Ile Pro Lys Asp Gln Trp Met Lys Lys Trp Trp Glu Met
465 470 475 480
Lys Arg Glu Ile Val Gly Val Val Glu Pro Val Pro His Asp Glu Thr
485 490 495
Tyr Cys Asp Pro Ala Ser Leu Phe His Val Ser Asn Asp Tyr Ser Phe
500 505 510
Ile Arg Tyr Tyr Thr Arg Thr Leu Tyr Gln Phe Gln Phe Gln Glu Ala
515 520 525
Leu Cys Gln Ala Ala Lys His Glu Gly Pro Leu His Lys Cys Asp Ile
530 535 540
Ser Asn Ser Thr Glu Ala Gly Gln Lys Leu Phe Asn Met Leu Arg Leu
545 550 555 560
Gly Lys Ser Glu Pro Trp Thr Leu Ala Leu Glu Asn Val Val Gly Ala
565 570 575
Lys Asn Met Asn Val Arg Pro Leu Leu Asn Tyr Phe Glu Pro Leu Phe
580 585 590
Thr Trp Leu Lys Asp Gln Asn Lys Asn Ser Phe Val Gly Trp Ser Thr
595 600 605
Asp Trp Ser Pro Tyr Ala Asp Ser Gln Asp Val Thr Val Pro Cys Arg
610 615 620
Val Pro Pro Pro Pro Pro Cys Cys His Pro Arg Leu Ser Leu His Arg
625 630 635 640
Pro Ala Leu Glu Asp Leu Leu Leu Gly Ser Glu Ala Asn Leu Thr Cys
645 650 655
Thr Leu Thr Gly Leu Arg Asp Ala Ser Gly Ala Thr Phe Thr Trp Thr
660 665 670
Pro Ser Ser Gly Lys Ser Ala Val Gln Gly Pro Pro Glu Arg Asp Leu
675 680 685
Cys Gly Cys Tyr Ser Val Ser Ser Val Leu Pro Gly Cys Ala Gln Pro
690 695 700
Trp Asn His Gly Glu Thr Phe Thr Cys Thr Ala Ala His Pro Glu Leu
705 710 715 720
Lys Thr Pro Leu Thr Ala Asn Ile Thr Lys Ser Gly Asn Thr Phe Arg
725 730 735
Pro Glu Val His Leu Leu Pro Pro Pro Ser Glu Glu Leu Ala Leu Asn
740 745 750
Glu Leu Val Thr Leu Thr Cys Leu Ala Arg Gly Phe Ser Pro Lys Asp
755 760 765
Val Leu Val Arg Trp Leu Gln Gly Ser Gln Glu Leu Pro Arg Glu Lys
770 775 780
Tyr Leu Thr Trp Ala Ser Arg Gln Glu Pro Ser Gln Gly Thr Thr Thr
785 790 795 800
Tyr Ala Val Thr Ser Ile Leu Arg Val Ala Ala Glu Asp Trp Lys Lys
805 810 815
Gly Glu Thr Phe Ser Cys Met Val Gly His Glu Ala Leu Pro Leu Ala
820 825 830
Phe Thr Gln Lys Thr Ile Asp Arg Met Ala Gly Lys Pro Thr His Ile
835 840 845
Asn Val Ser Val Val Met Ala Glu Ala Asp Gly Thr Cys Tyr
850 855 860
<210> 14
<211> 2589
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 14
atgtcaagct cttcctggct ccttctcagc cttgttgctg taactgctgc tcagtccacc 60
attgaggaac aggccaagac atttttggac aagtttaacc acgaagccga agacctgttc 120
tatcaaagtt cacttgcttc ttggaattat aacaccaata ttactgaaga gaatgtccaa 180
aacatgaata atgctgggga caaatggtct gcctttttaa aggaacagtc cacacttgcc 240
caaatgtatc cactacaaga aattcagaat ctcacagtca agcttcagct gcaggctctt 300
cagcaaaatg ggtcttcagt gctctcagaa gacaagagca aacggttgaa cacaattcta 360
aatacaatga gcaccatcta cagtactgga aaagtttgta acccagataa tccacaagaa 420
tgcttattac ttgaaccagg tttgaatgaa ataatggcaa acagtttaga ctacaatgag 480
aggctctggg cttgggaaag ctggagatct gaggtcggca agcagctgag gccattatat 540
gaagagtatg tggtcttgaa aaatgagatg gcaagagcaa atcattatga ggactatggg 600
gattattgga gaggagacta tgaagtaaat ggggtagatg gctatgacta cagccgcggc 660
cagttgattg aagatgtgga acataccttt gaagagatta aaccattata tgaacatctt 720
catgcctatg tgagggcaaa gttgatgaat gcctatcctt cctatatcag tccaattgga 780
tgcctccctg ctcatttgct tggtgatatg tggggtagat tttggacaaa tctgtactct 840
ttgacagttc cctttggaca gaaaccaaac atagatgtta ctgatgcaat ggtggaccag 900
gcctgggatg cacagagaat attcaaggag gccgagaagt tctttgtatc tgttggtctt 960
cctaatatga ctcaaggatt ctgggaaaat tccatgctaa cggacccagg aaatgttcag 1020
aaagcagtct gccatcccac agcttgggac ctggggaagg gcgacttcag gatccttatg 1080
tgcacaaagg tgacaatgga cgacttcctg acagctcatc atgagatggg gcatatccag 1140
tatgatatgg catatgctgc acaacctttt ctgctaagaa atggagctaa tgaaggattc 1200
catgaagctg ttggggaaat catgtcactt tctgcagcca cacctaagca tttaaaatcc 1260
attggtcttc tgtcacccga ttttcaagaa gacaatgaaa cagaaataaa cttcctgctc 1320
aaacaagcac tcacgattgt tgggactctg ccatttactt acatgttaga gaagtggagg 1380
tggatggtct ttaaagggga aattcccaaa gaccagtgga tgaaaaagtg gtgggagatg 1440
aagcgagaga tagttggggt ggtggaacct gtgccccatg atgaaacata ctgtgacccc 1500
gcatctctgt tccatgtttc taatgattac tcattcattc gatattacac aaggaccctt 1560
taccaattcc agtttcaaga agcactttgt caagcagcta aacatgaagg ccctctgcac 1620
aaatgtgaca tctcaaactc tacagaagct ggacagaaac tgttcaatat gctgaggctt 1680
ggaaaatcag aaccctggac cctagcattg gaaaatgttg taggagcaaa gaacatgaat 1740
gtaaggccac tgctcaacta ctttgagccc ttatttacct ggctgaaaga ccagaacaag 1800
aattcttttg tgggatggag taccgactgg agtccatatg cagacagcca ggatgtgact 1860
gtgccctgcc gagttccccc acctccccca tgctgccacc cccgactgtc gctgcaccga 1920
ccggccctcg aggacctgct cttaggttca gaagcgaacc tcacgtgcac actgaccggc 1980
ctgagagatg cctctggtgc caccttcacc tggacgccct caagtgggaa gagcgctgtt 2040
caaggaccac ctgagcgtga cctctgtggc tgctacagcg tgtccagtgt cctgcctggc 2100
tgtgcccagc catggaacca tggggagacc ttcacctgca ctgctgccca ccccgagttg 2160
aagaccccac taaccgccaa catcacaaaa tccggaaaca cattccggcc cgaggtccac 2220
ctgctgccgc cgccgtcgga ggagctggcc ctgaacgagc tggtgacgct gacgtgcctg 2280
gcacgtggct tcagccccaa ggatgtgctg gttcgctggc tgcaggggtc acaggagctg 2340
ccccgcgaga agtacctgac ttgggcatcc cggcaggagc ccagccaggg caccaccacc 2400
tatgctgtga ccagcatact gcgcgtggca gccgaggact ggaagaaggg ggaaaccttc 2460
tcctgcatgg tgggccacga ggccctgccg ctggccttca cacagaagac catcgaccgc 2520
atggcgggta aacccaccca tatcaatgtg tctgttgtca tggcggaggc ggacggcacc 2580
tgctactga 2589
<210> 15
<211> 830
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 15
Gln Ser Thr Ile Glu Glu Gln Ala Lys Thr Phe Leu Asp Lys Phe Asn
1 5 10 15
His Glu Ala Glu Asp Leu Phe Tyr Gln Ser Ser Leu Ala Ser Trp Asn
20 25 30
Tyr Asn Thr Asn Ile Thr Glu Glu Asn Val Gln Asn Met Asn Asn Ala
35 40 45
Gly Asp Lys Trp Ser Ala Phe Leu Lys Glu Gln Ser Thr Leu Ala Gln
50 55 60
Met Tyr Pro Leu Gln Glu Ile Gln Asn Leu Thr Val Lys Leu Gln Leu
65 70 75 80
Gln Ala Leu Gln Gln Asn Gly Ser Ser Val Leu Ser Glu Asp Lys Ser
85 90 95
Lys Arg Leu Asn Thr Ile Leu Asn Thr Met Ser Thr Ile Tyr Ser Thr
100 105 110
Gly Lys Val Cys Asn Pro Asp Asn Pro Gln Glu Cys Leu Leu Leu Glu
115 120 125
Pro Gly Leu Asn Glu Ile Met Ala Asn Ser Leu Asp Tyr Asn Glu Arg
130 135 140
Leu Trp Ala Trp Glu Ser Trp Arg Ser Glu Val Gly Lys Gln Leu Arg
145 150 155 160
Pro Leu Tyr Glu Glu Tyr Val Val Leu Lys Asn Glu Met Ala Arg Ala
165 170 175
Asn His Tyr Glu Asp Tyr Gly Asp Tyr Trp Arg Gly Asp Tyr Glu Val
180 185 190
Asn Gly Val Asp Gly Tyr Asp Tyr Ser Arg Gly Gln Leu Ile Glu Asp
195 200 205
Val Glu His Thr Phe Glu Glu Ile Lys Pro Leu Tyr Glu His Leu His
210 215 220
Ala Tyr Val Arg Ala Lys Leu Met Asn Ala Tyr Pro Ser Tyr Ile Ser
225 230 235 240
Pro Ile Gly Cys Leu Pro Ala His Leu Leu Gly Asp Met Trp Gly Arg
245 250 255
Phe Trp Thr Asn Leu Tyr Ser Leu Thr Val Pro Phe Gly Gln Lys Pro
260 265 270
Asn Ile Asp Val Thr Asp Ala Met Val Asp Gln Ala Trp Asp Ala Gln
275 280 285
Arg Ile Phe Lys Glu Ala Glu Lys Phe Phe Val Ser Val Gly Leu Pro
290 295 300
Asn Met Thr Gln Gly Phe Trp Glu Asn Ser Met Leu Thr Asp Pro Gly
305 310 315 320
Asn Val Gln Lys Ala Val Cys His Pro Thr Ala Trp Asp Leu Gly Lys
325 330 335
Gly Asp Phe Arg Ile Leu Met Cys Thr Lys Val Thr Met Asp Asp Phe
340 345 350
Leu Thr Ala His His Glu Met Gly His Ile Gln Tyr Asp Met Ala Tyr
355 360 365
Ala Ala Gln Pro Phe Leu Leu Arg Asn Gly Ala Asn Glu Gly Phe His
370 375 380
Glu Ala Val Gly Glu Ile Met Ser Leu Ser Ala Ala Thr Pro Lys His
385 390 395 400
Leu Lys Ser Ile Gly Leu Leu Ser Pro Asp Phe Gln Glu Asp Asn Glu
405 410 415
Thr Glu Ile Asn Phe Leu Leu Lys Gln Ala Leu Thr Ile Val Gly Thr
420 425 430
Leu Pro Phe Thr Tyr Met Leu Glu Lys Trp Arg Trp Met Val Phe Lys
435 440 445
Gly Glu Ile Pro Lys Asp Gln Trp Met Lys Lys Trp Trp Glu Met Lys
450 455 460
Arg Glu Ile Val Gly Val Val Glu Pro Val Pro His Asp Glu Thr Tyr
465 470 475 480
Cys Asp Pro Ala Ser Leu Phe His Val Ser Asn Asp Tyr Ser Phe Ile
485 490 495
Arg Tyr Tyr Thr Arg Thr Leu Tyr Gln Phe Gln Phe Gln Glu Ala Leu
500 505 510
Cys Gln Ala Ala Lys His Glu Gly Pro Leu His Lys Cys Asp Ile Ser
515 520 525
Asn Ser Thr Glu Ala Gly Gln Lys Leu Phe Asn Met Leu Arg Leu Gly
530 535 540
Lys Ser Glu Pro Trp Thr Leu Ala Leu Glu Asn Val Val Gly Ala Lys
545 550 555 560
Asn Met Asn Val Arg Pro Leu Leu Asn Tyr Phe Glu Pro Leu Phe Thr
565 570 575
Trp Leu Lys Asp Gln Asn Lys Asn Ser Phe Val Gly Trp Ser Thr Asp
580 585 590
Trp Ser Pro Tyr Ala Asp Glu Pro Lys Ser Ser Asp Lys Thr His Thr
595 600 605
Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe
610 615 620
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
625 630 635 640
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
645 650 655
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
660 665 670
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
675 680 685
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
690 695 700
Ala Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
705 710 715 720
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
725 730 735
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
740 745 750
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
755 760 765
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
770 775 780
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
785 790 795 800
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
805 810 815
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
820 825 830
<210> 16
<211> 827
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 16
Gln Ser Thr Ile Glu Glu Gln Ala Lys Thr Phe Leu Asp Lys Phe Asn
1 5 10 15
His Glu Ala Glu Asp Leu Phe Tyr Gln Ser Ser Leu Ala Ser Trp Asn
20 25 30
Tyr Asn Thr Asn Ile Thr Glu Glu Asn Val Gln Asn Met Asn Asn Ala
35 40 45
Gly Asp Lys Trp Ser Ala Phe Leu Lys Glu Gln Ser Thr Leu Ala Gln
50 55 60
Met Tyr Pro Leu Gln Glu Ile Gln Asn Leu Thr Val Lys Leu Gln Leu
65 70 75 80
Gln Ala Leu Gln Gln Asn Gly Ser Ser Val Leu Ser Glu Asp Lys Ser
85 90 95
Lys Arg Leu Asn Thr Ile Leu Asn Thr Met Ser Thr Ile Tyr Ser Thr
100 105 110
Gly Lys Val Cys Asn Pro Asp Asn Pro Gln Glu Cys Leu Leu Leu Glu
115 120 125
Pro Gly Leu Asn Glu Ile Met Ala Asn Ser Leu Asp Tyr Asn Glu Arg
130 135 140
Leu Trp Ala Trp Glu Ser Trp Arg Ser Glu Val Gly Lys Gln Leu Arg
145 150 155 160
Pro Leu Tyr Glu Glu Tyr Val Val Leu Lys Asn Glu Met Ala Arg Ala
165 170 175
Asn His Tyr Glu Asp Tyr Gly Asp Tyr Trp Arg Gly Asp Tyr Glu Val
180 185 190
Asn Gly Val Asp Gly Tyr Asp Tyr Ser Arg Gly Gln Leu Ile Glu Asp
195 200 205
Val Glu His Thr Phe Glu Glu Ile Lys Pro Leu Tyr Glu His Leu His
210 215 220
Ala Tyr Val Arg Ala Lys Leu Met Asn Ala Tyr Pro Ser Tyr Ile Ser
225 230 235 240
Pro Ile Gly Cys Leu Pro Ala His Leu Leu Gly Asp Met Trp Gly Arg
245 250 255
Phe Trp Thr Asn Leu Tyr Ser Leu Thr Val Pro Phe Gly Gln Lys Pro
260 265 270
Asn Ile Asp Val Thr Asp Ala Met Val Asp Gln Ala Trp Asp Ala Gln
275 280 285
Arg Ile Phe Lys Glu Ala Glu Lys Phe Phe Val Ser Val Gly Leu Pro
290 295 300
Asn Met Thr Gln Gly Phe Trp Glu Asn Ser Met Leu Thr Asp Pro Gly
305 310 315 320
Asn Val Gln Lys Ala Val Cys His Pro Thr Ala Trp Asp Leu Gly Lys
325 330 335
Gly Asp Phe Arg Ile Leu Met Cys Thr Lys Val Thr Met Asp Asp Phe
340 345 350
Leu Thr Ala His His Glu Met Gly His Ile Gln Tyr Asp Met Ala Tyr
355 360 365
Ala Ala Gln Pro Phe Leu Leu Arg Asn Gly Ala Asn Glu Gly Phe His
370 375 380
Glu Ala Val Gly Glu Ile Met Ser Leu Ser Ala Ala Thr Pro Lys His
385 390 395 400
Leu Lys Ser Ile Gly Leu Leu Ser Pro Asp Phe Gln Glu Asp Asn Glu
405 410 415
Thr Glu Ile Asn Phe Leu Leu Lys Gln Ala Leu Thr Ile Val Gly Thr
420 425 430
Leu Pro Phe Thr Tyr Met Leu Glu Lys Trp Arg Trp Met Val Phe Lys
435 440 445
Gly Glu Ile Pro Lys Asp Gln Trp Met Lys Lys Trp Trp Glu Met Lys
450 455 460
Arg Glu Ile Val Gly Val Val Glu Pro Val Pro His Asp Glu Thr Tyr
465 470 475 480
Cys Asp Pro Ala Ser Leu Phe His Val Ser Asn Asp Tyr Ser Phe Ile
485 490 495
Arg Tyr Tyr Thr Arg Thr Leu Tyr Gln Phe Gln Phe Gln Glu Ala Leu
500 505 510
Cys Gln Ala Ala Lys His Glu Gly Pro Leu His Lys Cys Asp Ile Ser
515 520 525
Asn Ser Thr Glu Ala Gly Gln Lys Leu Phe Asn Met Leu Arg Leu Gly
530 535 540
Lys Ser Glu Pro Trp Thr Leu Ala Leu Glu Asn Val Val Gly Ala Lys
545 550 555 560
Asn Met Asn Val Arg Pro Leu Leu Asn Tyr Phe Glu Pro Leu Phe Thr
565 570 575
Trp Leu Lys Asp Gln Asn Lys Asn Ser Phe Val Gly Trp Ser Thr Asp
580 585 590
Trp Ser Pro Tyr Ala Asp Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro
595 600 605
Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro
610 615 620
Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr
625 630 635 640
Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn
645 650 655
Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg
660 665 670
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val
675 680 685
Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
690 695 700
Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys
705 710 715 720
Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu
725 730 735
Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe
740 745 750
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu
755 760 765
Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe
770 775 780
Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly
785 790 795 800
Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr
805 810 815
Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys
820 825
<210> 17
<211> 858
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 17
Gln Ser Thr Ile Glu Glu Gln Ala Lys Thr Phe Leu Asp Lys Phe Asn
1 5 10 15
His Glu Ala Glu Asp Leu Phe Tyr Gln Ser Ser Leu Ala Ser Trp Asn
20 25 30
Tyr Asn Thr Asn Ile Thr Glu Glu Asn Val Gln Asn Met Asn Asn Ala
35 40 45
Gly Asp Lys Trp Ser Ala Phe Leu Lys Glu Gln Ser Thr Leu Ala Gln
50 55 60
Met Tyr Pro Leu Gln Glu Ile Gln Asn Leu Thr Val Lys Leu Gln Leu
65 70 75 80
Gln Ala Leu Gln Gln Asn Gly Ser Ser Val Leu Ser Glu Asp Lys Ser
85 90 95
Lys Arg Leu Asn Thr Ile Leu Asn Thr Met Ser Thr Ile Tyr Ser Thr
100 105 110
Gly Lys Val Cys Asn Pro Asp Asn Pro Gln Glu Cys Leu Leu Leu Glu
115 120 125
Pro Gly Leu Asn Glu Ile Met Ala Asn Ser Leu Asp Tyr Asn Glu Arg
130 135 140
Leu Trp Ala Trp Glu Ser Trp Arg Ser Glu Val Gly Lys Gln Leu Arg
145 150 155 160
Pro Leu Tyr Glu Glu Tyr Val Val Leu Lys Asn Glu Met Ala Arg Ala
165 170 175
Asn His Tyr Glu Asp Tyr Gly Asp Tyr Trp Arg Gly Asp Tyr Glu Val
180 185 190
Asn Gly Val Asp Gly Tyr Asp Tyr Ser Arg Gly Gln Leu Ile Glu Asp
195 200 205
Val Glu His Thr Phe Glu Glu Ile Lys Pro Leu Tyr Glu His Leu His
210 215 220
Ala Tyr Val Arg Ala Lys Leu Met Asn Ala Tyr Pro Ser Tyr Ile Ser
225 230 235 240
Pro Ile Gly Cys Leu Pro Ala His Leu Leu Gly Asp Met Trp Gly Arg
245 250 255
Phe Trp Thr Asn Leu Tyr Ser Leu Thr Val Pro Phe Gly Gln Lys Pro
260 265 270
Asn Ile Asp Val Thr Asp Ala Met Val Asp Gln Ala Trp Asp Ala Gln
275 280 285
Arg Ile Phe Lys Glu Ala Glu Lys Phe Phe Val Ser Val Gly Leu Pro
290 295 300
Asn Met Thr Gln Gly Phe Trp Glu Asn Ser Met Leu Thr Asp Pro Gly
305 310 315 320
Asn Val Gln Lys Ala Val Cys His Pro Thr Ala Trp Asp Leu Gly Lys
325 330 335
Gly Asp Phe Arg Ile Leu Met Cys Thr Lys Val Thr Met Asp Asp Phe
340 345 350
Leu Thr Ala His His Glu Met Gly His Ile Gln Tyr Asp Met Ala Tyr
355 360 365
Ala Ala Gln Pro Phe Leu Leu Arg Asn Gly Ala Asn Glu Gly Phe His
370 375 380
Glu Ala Val Gly Glu Ile Met Ser Leu Ser Ala Ala Thr Pro Lys His
385 390 395 400
Leu Lys Ser Ile Gly Leu Leu Ser Pro Asp Phe Gln Glu Asp Asn Glu
405 410 415
Thr Glu Ile Asn Phe Leu Leu Lys Gln Ala Leu Thr Ile Val Gly Thr
420 425 430
Leu Pro Phe Thr Tyr Met Leu Glu Lys Trp Arg Trp Met Val Phe Lys
435 440 445
Gly Glu Ile Pro Lys Asp Gln Trp Met Lys Lys Trp Trp Glu Met Lys
450 455 460
Arg Glu Ile Val Gly Val Val Glu Pro Val Pro His Asp Glu Thr Tyr
465 470 475 480
Cys Asp Pro Ala Ser Leu Phe His Val Ser Asn Asp Tyr Ser Phe Ile
485 490 495
Arg Tyr Tyr Thr Arg Thr Leu Tyr Gln Phe Gln Phe Gln Glu Ala Leu
500 505 510
Cys Gln Ala Ala Lys His Glu Gly Pro Leu His Lys Cys Asp Ile Ser
515 520 525
Asn Ser Thr Glu Ala Gly Gln Lys Leu Phe Asn Met Leu Arg Leu Gly
530 535 540
Lys Ser Glu Pro Trp Thr Leu Ala Leu Glu Asn Val Val Gly Ala Lys
545 550 555 560
Asn Met Asn Val Arg Pro Leu Leu Asn Tyr Phe Glu Pro Leu Phe Thr
565 570 575
Trp Leu Lys Asp Gln Asn Lys Asn Ser Phe Val Gly Trp Ser Thr Asp
580 585 590
Trp Ser Pro Tyr Ala Asp Ser Gln Asp Val Thr Val Pro Cys Pro Val
595 600 605
Pro Ser Thr Pro Pro Thr Pro Ser Pro Ser Thr Pro Pro Thr Pro Ser
610 615 620
Pro Ser Cys Cys His Pro Arg Leu Ser Leu His Arg Pro Ala Leu Glu
625 630 635 640
Asp Leu Leu Leu Gly Ser Glu Ala Asn Leu Thr Cys Thr Leu Thr Gly
645 650 655
Leu Arg Asp Ala Ser Gly Val Thr Phe Thr Trp Thr Pro Ser Ser Gly
660 665 670
Lys Ser Ala Val Gln Gly Pro Pro Glu Arg Asp Leu Cys Gly Cys Tyr
675 680 685
Ser Val Ser Ser Val Leu Pro Gly Cys Ala Glu Pro Trp Asn His Gly
690 695 700
Lys Thr Phe Thr Cys Thr Ala Ala Tyr Pro Glu Ser Lys Thr Pro Leu
705 710 715 720
Thr Ala Thr Leu Ser Lys Ser Gly Asn Thr Phe Arg Pro Glu Val His
725 730 735
Leu Leu Pro Pro Pro Ser Glu Glu Leu Ala Leu Asn Glu Leu Val Thr
740 745 750
Leu Thr Cys Leu Ala Arg Gly Phe Ser Pro Lys Asp Val Leu Val Arg
755 760 765
Trp Leu Gln Gly Ser Gln Glu Leu Pro Arg Glu Lys Tyr Leu Thr Trp
770 775 780
Ala Ser Arg Gln Glu Pro Ser Gln Gly Thr Thr Thr Phe Ala Val Thr
785 790 795 800
Ser Ile Leu Arg Val Ala Ala Glu Asp Trp Lys Lys Gly Asp Thr Phe
805 810 815
Ser Cys Met Val Gly His Glu Ala Leu Pro Leu Ala Phe Thr Gln Lys
820 825 830
Thr Ile Asp Arg Leu Ala Gly Lys Pro Thr His Val Asn Val Ser Val
835 840 845
Val Met Ala Glu Val Asp Gly Thr Cys Tyr
850 855
<210> 18
<211> 845
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 18
Gln Ser Thr Ile Glu Glu Gln Ala Lys Thr Phe Leu Asp Lys Phe Asn
1 5 10 15
His Glu Ala Glu Asp Leu Phe Tyr Gln Ser Ser Leu Ala Ser Trp Asn
20 25 30
Tyr Asn Thr Asn Ile Thr Glu Glu Asn Val Gln Asn Met Asn Asn Ala
35 40 45
Gly Asp Lys Trp Ser Ala Phe Leu Lys Glu Gln Ser Thr Leu Ala Gln
50 55 60
Met Tyr Pro Leu Gln Glu Ile Gln Asn Leu Thr Val Lys Leu Gln Leu
65 70 75 80
Gln Ala Leu Gln Gln Asn Gly Ser Ser Val Leu Ser Glu Asp Lys Ser
85 90 95
Lys Arg Leu Asn Thr Ile Leu Asn Thr Met Ser Thr Ile Tyr Ser Thr
100 105 110
Gly Lys Val Cys Asn Pro Asp Asn Pro Gln Glu Cys Leu Leu Leu Glu
115 120 125
Pro Gly Leu Asn Glu Ile Met Ala Asn Ser Leu Asp Tyr Asn Glu Arg
130 135 140
Leu Trp Ala Trp Glu Ser Trp Arg Ser Glu Val Gly Lys Gln Leu Arg
145 150 155 160
Pro Leu Tyr Glu Glu Tyr Val Val Leu Lys Asn Glu Met Ala Arg Ala
165 170 175
Asn His Tyr Glu Asp Tyr Gly Asp Tyr Trp Arg Gly Asp Tyr Glu Val
180 185 190
Asn Gly Val Asp Gly Tyr Asp Tyr Ser Arg Gly Gln Leu Ile Glu Asp
195 200 205
Val Glu His Thr Phe Glu Glu Ile Lys Pro Leu Tyr Glu His Leu His
210 215 220
Ala Tyr Val Arg Ala Lys Leu Met Asn Ala Tyr Pro Ser Tyr Ile Ser
225 230 235 240
Pro Ile Gly Cys Leu Pro Ala His Leu Leu Gly Asp Met Trp Gly Arg
245 250 255
Phe Trp Thr Asn Leu Tyr Ser Leu Thr Val Pro Phe Gly Gln Lys Pro
260 265 270
Asn Ile Asp Val Thr Asp Ala Met Val Asp Gln Ala Trp Asp Ala Gln
275 280 285
Arg Ile Phe Lys Glu Ala Glu Lys Phe Phe Val Ser Val Gly Leu Pro
290 295 300
Asn Met Thr Gln Gly Phe Trp Glu Asn Ser Met Leu Thr Asp Pro Gly
305 310 315 320
Asn Val Gln Lys Ala Val Cys His Pro Thr Ala Trp Asp Leu Gly Lys
325 330 335
Gly Asp Phe Arg Ile Leu Met Cys Thr Lys Val Thr Met Asp Asp Phe
340 345 350
Leu Thr Ala His His Glu Met Gly His Ile Gln Tyr Asp Met Ala Tyr
355 360 365
Ala Ala Gln Pro Phe Leu Leu Arg Asn Gly Ala Asn Glu Gly Phe His
370 375 380
Glu Ala Val Gly Glu Ile Met Ser Leu Ser Ala Ala Thr Pro Lys His
385 390 395 400
Leu Lys Ser Ile Gly Leu Leu Ser Pro Asp Phe Gln Glu Asp Asn Glu
405 410 415
Thr Glu Ile Asn Phe Leu Leu Lys Gln Ala Leu Thr Ile Val Gly Thr
420 425 430
Leu Pro Phe Thr Tyr Met Leu Glu Lys Trp Arg Trp Met Val Phe Lys
435 440 445
Gly Glu Ile Pro Lys Asp Gln Trp Met Lys Lys Trp Trp Glu Met Lys
450 455 460
Arg Glu Ile Val Gly Val Val Glu Pro Val Pro His Asp Glu Thr Tyr
465 470 475 480
Cys Asp Pro Ala Ser Leu Phe His Val Ser Asn Asp Tyr Ser Phe Ile
485 490 495
Arg Tyr Tyr Thr Arg Thr Leu Tyr Gln Phe Gln Phe Gln Glu Ala Leu
500 505 510
Cys Gln Ala Ala Lys His Glu Gly Pro Leu His Lys Cys Asp Ile Ser
515 520 525
Asn Ser Thr Glu Ala Gly Gln Lys Leu Phe Asn Met Leu Arg Leu Gly
530 535 540
Lys Ser Glu Pro Trp Thr Leu Ala Leu Glu Asn Val Val Gly Ala Lys
545 550 555 560
Asn Met Asn Val Arg Pro Leu Leu Asn Tyr Phe Glu Pro Leu Phe Thr
565 570 575
Trp Leu Lys Asp Gln Asn Lys Asn Ser Phe Val Gly Trp Ser Thr Asp
580 585 590
Trp Ser Pro Tyr Ala Asp Ser Gln Asp Val Thr Val Pro Cys Arg Val
595 600 605
Pro Pro Pro Pro Pro Cys Cys His Pro Arg Leu Ser Leu His Arg Pro
610 615 620
Ala Leu Glu Asp Leu Leu Leu Gly Ser Glu Ala Asn Leu Thr Cys Thr
625 630 635 640
Leu Thr Gly Leu Arg Asp Ala Ser Gly Ala Thr Phe Thr Trp Thr Pro
645 650 655
Ser Ser Gly Lys Ser Ala Val Gln Gly Pro Pro Glu Arg Asp Leu Cys
660 665 670
Gly Cys Tyr Ser Val Ser Ser Val Leu Pro Gly Cys Ala Gln Pro Trp
675 680 685
Asn His Gly Glu Thr Phe Thr Cys Thr Ala Ala His Pro Glu Leu Lys
690 695 700
Thr Pro Leu Thr Ala Asn Ile Thr Lys Ser Gly Asn Thr Phe Arg Pro
705 710 715 720
Glu Val His Leu Leu Pro Pro Pro Ser Glu Glu Leu Ala Leu Asn Glu
725 730 735
Leu Val Thr Leu Thr Cys Leu Ala Arg Gly Phe Ser Pro Lys Asp Val
740 745 750
Leu Val Arg Trp Leu Gln Gly Ser Gln Glu Leu Pro Arg Glu Lys Tyr
755 760 765
Leu Thr Trp Ala Ser Arg Gln Glu Pro Ser Gln Gly Thr Thr Thr Tyr
770 775 780
Ala Val Thr Ser Ile Leu Arg Val Ala Ala Glu Asp Trp Lys Lys Gly
785 790 795 800
Glu Thr Phe Ser Cys Met Val Gly His Glu Ala Leu Pro Leu Ala Phe
805 810 815
Thr Gln Lys Thr Ile Asp Arg Met Ala Gly Lys Pro Thr His Ile Asn
820 825 830
Val Ser Val Val Met Ala Glu Ala Asp Gly Thr Cys Tyr
835 840 845
<210> 19
<211> 847
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 19
Met Ser Ser Ser Ser Trp Leu Leu Leu Ser Leu Val Ala Val Thr Ala
1 5 10 15
Ala Gln Ser Thr Ile Glu Glu Gln Ala Lys Thr Phe Leu Asp Lys Phe
20 25 30
Asn His Glu Ala Glu Asp Leu Phe Tyr Gln Ser Ser Leu Ala Ser Trp
35 40 45
Asn Tyr Asn Thr Asn Ile Thr Glu Glu Asn Val Gln Asn Met Asn Asn
50 55 60
Ala Gly Asp Lys Trp Ser Ala Phe Leu Lys Glu Gln Ser Thr Leu Ala
65 70 75 80
Gln Met Tyr Pro Leu Gln Glu Ile Gln Asn Leu Thr Val Lys Leu Gln
85 90 95
Leu Gln Ala Leu Gln Gln Asn Gly Ser Ser Val Leu Ser Glu Asp Lys
100 105 110
Ser Lys Arg Leu Asn Thr Ile Leu Asn Thr Met Ser Thr Ile Tyr Ser
115 120 125
Thr Gly Lys Val Cys Asn Pro Asp Asn Pro Gln Glu Cys Leu Leu Leu
130 135 140
Glu Pro Gly Leu Asn Glu Ile Met Ala Asn Ser Leu Asp Tyr Asn Glu
145 150 155 160
Arg Leu Trp Ala Trp Glu Ser Trp Arg Ser Glu Val Gly Lys Gln Leu
165 170 175
Arg Pro Leu Tyr Glu Glu Tyr Val Val Leu Lys Asn Glu Met Ala Arg
180 185 190
Ala Asn His Tyr Glu Asp Tyr Gly Asp Tyr Trp Arg Gly Asp Tyr Glu
195 200 205
Val Asn Gly Val Asp Gly Tyr Asp Tyr Ser Arg Gly Gln Leu Ile Glu
210 215 220
Asp Val Glu His Thr Phe Glu Glu Ile Lys Pro Leu Tyr Glu His Leu
225 230 235 240
His Ala Tyr Val Arg Ala Lys Leu Met Asn Ala Tyr Pro Ser Tyr Ile
245 250 255
Ser Pro Ile Gly Cys Leu Pro Ala His Leu Leu Gly Asp Met Trp Gly
260 265 270
Arg Phe Trp Thr Asn Leu Tyr Ser Leu Thr Val Pro Phe Gly Gln Lys
275 280 285
Pro Asn Ile Asp Val Thr Asp Ala Met Val Asp Gln Ala Trp Asp Ala
290 295 300
Gln Arg Ile Phe Lys Glu Ala Glu Lys Phe Phe Val Ser Val Gly Leu
305 310 315 320
Pro Asn Met Thr Gln Gly Phe Trp Glu Asn Ser Met Leu Thr Asp Pro
325 330 335
Gly Asn Val Gln Lys Ala Val Cys His Pro Thr Ala Trp Asp Leu Gly
340 345 350
Lys Gly Asp Phe Arg Ile Leu Met Cys Thr Lys Val Thr Met Asp Asp
355 360 365
Phe Leu Thr Ala His His Glu Met Gly His Ile Gln Tyr Asp Met Ala
370 375 380
Tyr Ala Ala Gln Pro Phe Leu Leu Arg Asn Gly Ala Asn Glu Gly Phe
385 390 395 400
His Glu Ala Val Gly Glu Ile Met Ser Leu Ser Ala Ala Thr Pro Lys
405 410 415
His Leu Lys Ser Ile Gly Leu Leu Ser Pro Asp Phe Gln Glu Asp Asn
420 425 430
Glu Thr Glu Ile Asn Phe Leu Leu Lys Gln Ala Leu Thr Ile Val Gly
435 440 445
Thr Leu Pro Phe Thr Tyr Met Leu Glu Lys Trp Arg Trp Met Val Phe
450 455 460
Lys Gly Glu Ile Pro Lys Asp Gln Trp Met Lys Lys Trp Trp Glu Met
465 470 475 480
Lys Arg Glu Ile Val Gly Val Val Glu Pro Val Pro His Asp Glu Thr
485 490 495
Tyr Cys Asp Pro Ala Ser Leu Phe His Val Ser Asn Asp Tyr Ser Phe
500 505 510
Ile Arg Tyr Tyr Thr Arg Thr Leu Tyr Gln Phe Gln Phe Gln Glu Ala
515 520 525
Leu Cys Gln Ala Ala Lys His Glu Gly Pro Leu His Lys Cys Asp Ile
530 535 540
Ser Asn Ser Thr Glu Ala Gly Gln Lys Leu Phe Asn Met Leu Arg Leu
545 550 555 560
Gly Lys Ser Glu Pro Trp Thr Leu Ala Leu Glu Asn Val Val Gly Ala
565 570 575
Lys Asn Met Asn Val Arg Pro Leu Leu Asn Tyr Phe Glu Pro Leu Phe
580 585 590
Thr Trp Leu Lys Asp Gln Asn Lys Asn Ser Phe Val Gly Trp Ser Thr
595 600 605
Asp Trp Ser Pro Tyr Ala Asp Glu Pro Lys Ser Ser Asp Lys Thr His
610 615 620
Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val
625 630 635 640
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
645 650 655
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
660 665 670
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
675 680 685
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
690 695 700
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
705 710 715 720
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
725 730 735
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
740 745 750
Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu
755 760 765
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
770 775 780
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
785 790 795 800
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
805 810 815
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
820 825 830
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
835 840 845
<210> 20
<211> 2544
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 20
atgtcaagct cttcctggct ccttctcagc cttgttgctg taactgctgc tcagtccacc 60
attgaggaac aggccaagac atttttggac aagtttaacc acgaagccga agacctgttc 120
tatcaaagtt cacttgcttc ttggaattat aacaccaata ttactgaaga gaatgtccaa 180
aacatgaata atgctgggga caaatggtct gcctttttaa aggaacagtc cacacttgcc 240
caaatgtatc cactacaaga aattcagaat ctcacagtca agctccagct gcaggctctt 300
cagcaaaatg ggtcttcagt gctctcagaa gacaagagca aacggttgaa cacaattcta 360
aatacaatga gcaccatcta cagtactgga aaagtttgta acccagataa tccacaagaa 420
tgcttattac ttgaaccagg tttgaatgaa ataatggcaa acagtttaga ctacaatgag 480
aggctctggg cttgggaaag ctggagatct gaggtcggca agcagctgag gccattatat 540
gaagagtatg tggtcttgaa aaatgagatg gcaagagcaa atcattatga ggactatggg 600
gattattgga gaggagacta tgaagtaaat ggggtagatg gctatgacta cagccgcggc 660
cagttgattg aagatgtgga acataccttt gaagagatta aaccattata tgaacatctt 720
catgcctatg tgagggcaaa gttgatgaat gcctatcctt cctatatcag tccaattgga 780
tgcctccctg ctcatttgct tggtgatatg tggggtagat tttggacaaa tctgtactct 840
ttgacagttc cctttggaca gaaaccaaac atagatgtta ctgatgcaat ggtggaccag 900
gcctgggatg cacagagaat attcaaggag gccgagaagt tctttgtatc tgttggtctt 960
cctaatatga ctcaaggatt ctgggaaaat tccatgctaa cggacccagg aaatgttcag 1020
aaagcagtct gccatcccac agcttgggac ctggggaagg gcgacttcag gatccttatg 1080
tgcacaaagg tgacaatgga cgacttcctg acagctcatc atgagatggg gcatatccag 1140
tatgatatgg catatgctgc acaacctttt ctgctaagaa atggagctaa tgaaggattc 1200
catgaagctg ttggggaaat catgtcactt tctgcagcca cacctaagca tttaaaatcc 1260
attggtcttc tgtcacccga ttttcaagaa gacaatgaaa cagaaataaa cttcctgctc 1320
aaacaagcac tcacgattgt tgggactctg ccatttactt acatgttaga gaagtggagg 1380
tggatggtct ttaaagggga aattcccaaa gaccagtgga tgaaaaagtg gtgggagatg 1440
aagcgagaga tagttggggt ggtggaacct gtgccccatg atgaaacata ctgtgacccc 1500
gcatctctgt tccatgtttc taatgattac tcattcattc gatattacac aaggaccctt 1560
taccaattcc agtttcaaga agcactttgt caagcagcta aacatgaagg ccctctgcac 1620
aaatgtgaca tctcaaactc tacagaagct ggacagaaac tgttcaatat gctgaggctt 1680
ggaaaatcag aaccctggac cctagcattg gaaaatgttg taggagcaaa gaacatgaat 1740
gtaaggccac tgctcaacta ctttgagccc ttatttacct ggctgaaaga ccagaacaag 1800
aattcttttg tgggatggag taccgactgg agtccatatg cagacgagcc caaatcttcc 1860
gacaaaactc acacatgccc accgtgccca gcacctgaac tcctgggggg accgtcagtc 1920
ttcctcttcc ccccaaaacc caaggacacc ctcatgatct cccggacccc tgaggtcaca 1980
tgcgtggtgg tggacgtgag ccacgaagac cctgaggtca agttcaactg gtacgtggac 2040
ggcgtggagg tgcataatgc caagacaaag ccgcgggagg agcagtacaa cagcacgtac 2100
cgtgtggtca gcgtcctcac cgtcctgcac caggactggc tgaatggcaa ggagtacaag 2160
tgcaaggtct ccaacaaagc cctcccagcc cccatcgaga aaaccatctc caaagccaaa 2220
gggcagcccc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgaccaag 2280
aaccaggtca gcctgacctg cctggtcaaa ggcttctatc ccagcgacat cgccgtggag 2340
tgggagagca atgggcagcc ggagaacaac tacaagacca cgcctcccgt gctggactcc 2400
gacggctcct tcttcctcta tagcaagctc accgtggaca agagcaggtg gcagcagggg 2460
aacgtcttct catgctccgt gatgcatgag gctctgcaca accactacac gcagaagagc 2520
ctctccctgt ctccgggtaa atag 2544
<210> 21
<211> 972
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 21
Met Ser Ser Ser Ser Trp Leu Leu Leu Ser Leu Val Ala Val Thr Ala
1 5 10 15
Ala Gln Ser Thr Ile Glu Glu Gln Ala Lys Thr Phe Leu Asp Lys Phe
20 25 30
Asn His Glu Ala Glu Asp Leu Phe Tyr Gln Ser Ser Leu Ala Ser Trp
35 40 45
Asn Tyr Asn Thr Asn Ile Thr Glu Glu Asn Val Gln Asn Met Asn Asn
50 55 60
Ala Gly Asp Lys Trp Ser Ala Phe Leu Lys Glu Gln Ser Thr Leu Ala
65 70 75 80
Gln Met Tyr Pro Leu Gln Glu Ile Gln Asn Leu Thr Val Lys Leu Gln
85 90 95
Leu Gln Ala Leu Gln Gln Asn Gly Ser Ser Val Leu Ser Glu Asp Lys
100 105 110
Ser Lys Arg Leu Asn Thr Ile Leu Asn Thr Met Ser Thr Ile Tyr Ser
115 120 125
Thr Gly Lys Val Cys Asn Pro Asp Asn Pro Gln Glu Cys Leu Leu Leu
130 135 140
Glu Pro Gly Leu Asn Glu Ile Met Ala Asn Ser Leu Asp Tyr Asn Glu
145 150 155 160
Arg Leu Trp Ala Trp Glu Ser Trp Arg Ser Glu Val Gly Lys Gln Leu
165 170 175
Arg Pro Leu Tyr Glu Glu Tyr Val Val Leu Lys Asn Glu Met Ala Arg
180 185 190
Ala Asn His Tyr Glu Asp Tyr Gly Asp Tyr Trp Arg Gly Asp Tyr Glu
195 200 205
Val Asn Gly Val Asp Gly Tyr Asp Tyr Ser Arg Gly Gln Leu Ile Glu
210 215 220
Asp Val Glu His Thr Phe Glu Glu Ile Lys Pro Leu Tyr Glu His Leu
225 230 235 240
His Ala Tyr Val Arg Ala Lys Leu Met Asn Ala Tyr Pro Ser Tyr Ile
245 250 255
Ser Pro Ile Gly Cys Leu Pro Ala His Leu Leu Gly Asp Met Trp Gly
260 265 270
Arg Phe Trp Thr Asn Leu Tyr Ser Leu Thr Val Pro Phe Gly Gln Lys
275 280 285
Pro Asn Ile Asp Val Thr Asp Ala Met Val Asp Gln Ala Trp Asp Ala
290 295 300
Gln Arg Ile Phe Lys Glu Ala Glu Lys Phe Phe Val Ser Val Gly Leu
305 310 315 320
Pro Asn Met Thr Gln Gly Phe Trp Glu Asn Ser Met Leu Thr Asp Pro
325 330 335
Gly Asn Val Gln Lys Ala Val Cys His Pro Thr Ala Trp Asp Leu Gly
340 345 350
Lys Gly Asp Phe Arg Ile Leu Met Cys Thr Lys Val Thr Met Asp Asp
355 360 365
Phe Leu Thr Ala His His Glu Met Gly His Ile Gln Tyr Asp Met Ala
370 375 380
Tyr Ala Ala Gln Pro Phe Leu Leu Arg Asn Gly Ala Asn Glu Gly Phe
385 390 395 400
His Glu Ala Val Gly Glu Ile Met Ser Leu Ser Ala Ala Thr Pro Lys
405 410 415
His Leu Lys Ser Ile Gly Leu Leu Ser Pro Asp Phe Gln Glu Asp Asn
420 425 430
Glu Thr Glu Ile Asn Phe Leu Leu Lys Gln Ala Leu Thr Ile Val Gly
435 440 445
Thr Leu Pro Phe Thr Tyr Met Leu Glu Lys Trp Arg Trp Met Val Phe
450 455 460
Lys Gly Glu Ile Pro Lys Asp Gln Trp Met Lys Lys Trp Trp Glu Met
465 470 475 480
Lys Arg Glu Ile Val Gly Val Val Glu Pro Val Pro His Asp Glu Thr
485 490 495
Tyr Cys Asp Pro Ala Ser Leu Phe His Val Ser Asn Asp Tyr Ser Phe
500 505 510
Ile Arg Tyr Tyr Thr Arg Thr Leu Tyr Gln Phe Gln Phe Gln Glu Ala
515 520 525
Leu Cys Gln Ala Ala Lys His Glu Gly Pro Leu His Lys Cys Asp Ile
530 535 540
Ser Asn Ser Thr Glu Ala Gly Gln Lys Leu Phe Asn Met Leu Arg Leu
545 550 555 560
Gly Lys Ser Glu Pro Trp Thr Leu Ala Leu Glu Asn Val Val Gly Ala
565 570 575
Lys Asn Met Asn Val Arg Pro Leu Leu Asn Tyr Phe Glu Pro Leu Phe
580 585 590
Thr Trp Leu Lys Asp Gln Asn Lys Asn Ser Phe Val Gly Trp Ser Thr
595 600 605
Asp Trp Ser Pro Tyr Ala Asp Gln Ser Ile Lys Val Arg Ile Ser Leu
610 615 620
Lys Ser Ala Leu Gly Asp Lys Ala Tyr Glu Trp Asn Asp Asn Glu Met
625 630 635 640
Tyr Leu Phe Arg Ser Ser Val Ala Tyr Ala Met Arg Gln Tyr Phe Leu
645 650 655
Lys Val Lys Asn Gln Met Ile Leu Phe Gly Glu Glu Asp Val Arg Val
660 665 670
Ala Asn Leu Lys Pro Arg Ile Ser Phe Asn Phe Phe Val Thr Ala Pro
675 680 685
Lys Asn Val Ser Asp Ile Ile Pro Arg Thr Glu Val Glu Lys Ala Ile
690 695 700
Arg Met Ser Arg Ser Arg Ile Asn Asp Ala Phe Arg Leu Asn Asp Asn
705 710 715 720
Ser Leu Glu Phe Leu Gly Ile Gln Pro Thr Leu Gly Pro Pro Asn Gln
725 730 735
Pro Pro Val Ser Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro
740 745 750
Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe
755 760 765
Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
770 775 780
Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe
785 790 795 800
Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro
805 810 815
Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr
820 825 830
Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val
835 840 845
Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala
850 855 860
Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg
865 870 875 880
Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly
885 890 895
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro
900 905 910
Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser
915 920 925
Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln
930 935 940
Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His
945 950 955 960
Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
965 970
<210> 22
<211> 2919
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 22
atgtcaagct cttcctggct ccttctcagc cttgttgctg taactgctgc tcagtccacc 60
attgaggaac aggccaagac atttttggac aagtttaacc acgaagccga agacctgttc 120
tatcaaagtt cacttgcttc ttggaattat aacaccaata ttactgaaga gaatgtccaa 180
aacatgaata atgctgggga caaatggtct gcctttttaa aggaacagtc cacacttgcc 240
caaatgtatc cactacaaga aattcagaat ctcacagtca agctccagct gcaggctctt 300
cagcaaaatg ggtcttcagt gctctcagaa gacaagagca aacggttgaa cacaattcta 360
aatacaatga gcaccatcta cagtactgga aaagtttgta acccagataa tccacaagaa 420
tgcttattac ttgaaccagg tttgaatgaa ataatggcaa acagtttaga ctacaatgag 480
aggctctggg cttgggaaag ctggagatct gaggtcggca agcagctgag gccattatat 540
gaagagtatg tggtcttgaa aaatgagatg gcaagagcaa atcattatga ggactatggg 600
gattattgga gaggagacta tgaagtaaat ggggtagatg gctatgacta cagccgcggc 660
cagttgattg aagatgtgga acataccttt gaagagatta aaccattata tgaacatctt 720
catgcctatg tgagggcaaa gttgatgaat gcctatcctt cctatatcag tccaattgga 780
tgcctccctg ctcatttgct tggtgatatg tggggtagat tttggacaaa tctgtactct 840
ttgacagttc cctttggaca gaaaccaaac atagatgtta ctgatgcaat ggtggaccag 900
gcctgggatg cacagagaat attcaaggag gccgagaagt tctttgtatc tgttggtctt 960
cctaatatga ctcaaggatt ctgggaaaat tccatgctaa cggacccagg aaatgttcag 1020
aaagcagtct gccatcccac agcttgggac ctggggaagg gcgacttcag gatccttatg 1080
tgcacaaagg tgacaatgga cgacttcctg acagctcatc atgagatggg gcatatccag 1140
tatgatatgg catatgctgc acaacctttt ctgctaagaa atggagctaa tgaaggattc 1200
catgaagctg ttggggaaat catgtcactt tctgcagcca cacctaagca tttaaaatcc 1260
attggtcttc tgtcacccga ttttcaagaa gacaatgaaa cagaaataaa cttcctgctc 1320
aaacaagcac tcacgattgt tgggactctg ccatttactt acatgttaga gaagtggagg 1380
tggatggtct ttaaagggga aattcccaaa gaccagtgga tgaaaaagtg gtgggagatg 1440
aagcgagaga tagttggggt ggtggaacct gtgccccatg atgaaacata ctgtgacccc 1500
gcatctctgt tccatgtttc taatgattac tcattcattc gatattacac aaggaccctt 1560
taccaattcc agtttcaaga agcactttgt caagcagcta aacatgaagg ccctctgcac 1620
aaatgtgaca tctcaaactc tacagaagct ggacagaaac tgttcaatat gctgaggctt 1680
ggaaaatcag aaccctggac cctagcattg gaaaatgttg taggagcaaa gaacatgaat 1740
gtaaggccac tgctcaacta ctttgagccc ttatttacct ggctgaaaga ccagaacaag 1800
aattcttttg tgggatggag taccgactgg agtccatatg cagaccaaag catcaaagtg 1860
aggataagcc taaaatcagc tcttggagat aaagcatatg aatggaacga caatgaaatg 1920
tacctgttcc gatcatctgt tgcatatgct atgaggcagt actttttaaa agtaaaaaat 1980
cagatgattc tttttgggga ggaggatgtg cgagtggcta atttgaaacc aagaatctcc 2040
tttaatttct ttgtcactgc acctaaaaat gtgtctgata tcattcctag aactgaagtt 2100
gaaaaggcca tcaggatgtc ccggagccgt atcaatgatg ctttccgtct gaatgacaac 2160
agcctagagt ttctggggat acagccaaca cttggacctc ctaaccagcc ccctgtttcc 2220
gagcccaaat cttccgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg 2280
gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 2340
acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 2400
aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 2460
tacaacagca cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 2520
ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 2580
atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 2640
gatgagctga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 2700
gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 2760
cccgtgctgg actccgacgg ctccttcttc ctctatagca agctcaccgt ggacaagagc 2820
aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 2880
tacacgcaga agagcctctc cctgtctccg ggtaaatag 2919
<210> 23
<211> 621
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 23
Met Ser Ser Ser Ser Trp Leu Leu Leu Ser Leu Val Ala Val Thr Ala
1 5 10 15
Ala Gln Ser Thr Ile Glu Glu Gln Ala Lys Thr Phe Leu Asp Lys Phe
20 25 30
Asn His Glu Ala Glu Asp Leu Phe Tyr Gln Ser Ser Leu Ala Ser Trp
35 40 45
Asn Tyr Asn Thr Asn Ile Thr Glu Glu Asn Val Gln Asn Met Asn Asn
50 55 60
Ala Gly Asp Lys Trp Ser Ala Phe Leu Lys Glu Gln Ser Thr Leu Ala
65 70 75 80
Gln Met Tyr Pro Leu Gln Glu Ile Gln Asn Leu Thr Val Lys Leu Gln
85 90 95
Leu Gln Ala Leu Gln Gln Asn Gly Ser Ser Val Leu Ser Glu Asp Lys
100 105 110
Ser Lys Arg Leu Asn Thr Ile Leu Asn Thr Met Ser Thr Ile Tyr Ser
115 120 125
Thr Gly Lys Val Cys Asn Pro Asp Asn Pro Gln Glu Cys Leu Leu Leu
130 135 140
Glu Pro Gly Leu Asn Glu Ile Met Ala Asn Ser Leu Asp Tyr Asn Glu
145 150 155 160
Arg Leu Trp Ala Trp Glu Ser Trp Arg Ser Glu Val Gly Lys Gln Leu
165 170 175
Arg Pro Leu Tyr Glu Glu Tyr Val Val Leu Lys Asn Glu Met Ala Arg
180 185 190
Ala Asn His Tyr Glu Asp Tyr Gly Asp Tyr Trp Arg Gly Asp Tyr Glu
195 200 205
Val Asn Gly Val Asp Gly Tyr Asp Tyr Ser Arg Gly Gln Leu Ile Glu
210 215 220
Asp Val Glu His Thr Phe Glu Glu Ile Lys Pro Leu Tyr Glu His Leu
225 230 235 240
His Ala Tyr Val Arg Ala Lys Leu Met Asn Ala Tyr Pro Ser Tyr Ile
245 250 255
Ser Pro Ile Gly Cys Leu Pro Ala His Leu Leu Gly Asp Met Trp Gly
260 265 270
Arg Phe Trp Thr Asn Leu Tyr Ser Leu Thr Val Pro Phe Gly Gln Lys
275 280 285
Pro Asn Ile Asp Val Thr Asp Ala Met Val Asp Gln Ala Trp Asp Ala
290 295 300
Gln Arg Ile Phe Lys Glu Ala Glu Lys Phe Phe Val Ser Val Gly Leu
305 310 315 320
Pro Asn Met Thr Gln Gly Phe Trp Glu Asn Ser Met Leu Thr Asp Pro
325 330 335
Gly Asn Val Gln Lys Ala Val Cys His Pro Thr Ala Trp Asp Leu Gly
340 345 350
Lys Gly Asp Phe Arg Ile Leu Met Cys Thr Lys Val Thr Met Asp Asp
355 360 365
Phe Leu Thr Ala His His Glu Met Gly His Ile Gln Tyr Asp Met Ala
370 375 380
Tyr Ala Ala Gln Pro Phe Leu Leu Arg Asn Gly Ala Asn Glu Gly Phe
385 390 395 400
His Glu Ala Val Gly Glu Ile Met Ser Leu Ser Ala Ala Thr Pro Lys
405 410 415
His Leu Lys Ser Ile Gly Leu Leu Ser Pro Asp Phe Gln Glu Asp Asn
420 425 430
Glu Thr Glu Ile Asn Phe Leu Leu Lys Gln Ala Leu Thr Ile Val Gly
435 440 445
Thr Leu Pro Phe Thr Tyr Met Leu Glu Lys Trp Arg Trp Met Val Phe
450 455 460
Lys Gly Glu Ile Pro Lys Asp Gln Trp Met Lys Lys Trp Trp Glu Met
465 470 475 480
Lys Arg Glu Ile Val Gly Val Val Glu Pro Val Pro His Asp Glu Thr
485 490 495
Tyr Cys Asp Pro Ala Ser Leu Phe His Val Ser Asn Asp Tyr Ser Phe
500 505 510
Ile Arg Tyr Tyr Thr Arg Thr Leu Tyr Gln Phe Gln Phe Gln Glu Ala
515 520 525
Leu Cys Gln Ala Ala Lys His Glu Gly Pro Leu His Lys Cys Asp Ile
530 535 540
Ser Asn Ser Thr Glu Ala Gly Gln Lys Leu Phe Asn Met Leu Arg Leu
545 550 555 560
Gly Lys Ser Glu Pro Trp Thr Leu Ala Leu Glu Asn Val Val Gly Ala
565 570 575
Lys Asn Met Asn Val Arg Pro Leu Leu Asn Tyr Phe Glu Pro Leu Phe
580 585 590
Thr Trp Leu Lys Asp Gln Asn Lys Asn Ser Phe Val Gly Trp Ser Thr
595 600 605
Asp Trp Ser Pro Tyr Ala Asp His His His His His His
610 615 620
<210> 24
<211> 1863
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 24
atgtcaagct cttcctggct ccttctcagc cttgttgctg taactgctgc tcagtccacc 60
attgaggaac aggccaagac atttttggac aagtttaacc acgaagccga agacctgttc 120
tatcaaagtt cacttgcttc ttggaattat aacaccaata ttactgaaga gaatgtccaa 180
aacatgaata atgctgggga caaatggtct gcctttttaa aggaacagtc cacacttgcc 240
caaatgtatc cactacaaga aattcagaat ctcacagtca agcttcagct gcaggctctt 300
cagcaaaatg ggtcttcagt gctctcagaa gacaagagca aacggttgaa cacaattcta 360
aatacaatga gcaccatcta cagtactgga aaagtttgta acccagataa tccacaagaa 420
tgcttattac ttgaaccagg tttgaatgaa ataatggcaa acagtttaga ctacaatgag 480
aggctctggg cttgggaaag ctggagatct gaggtcggca agcagctgag gccattatat 540
gaagagtatg tggtcttgaa aaatgagatg gcaagagcaa atcattatga ggactatggg 600
gattattgga gaggagacta tgaagtaaat ggggtagatg gctatgacta cagccgcggc 660
cagttgattg aagatgtgga acataccttt gaagagatta aaccattata tgaacatctt 720
catgcctatg tgagggcaaa gttgatgaat gcctatcctt cctatatcag tccaattgga 780
tgcctccctg ctcatttgct tggtgatatg tggggtagat tttggacaaa tctgtactct 840
ttgacagttc cctttggaca gaaaccaaac atagatgtta ctgatgcaat ggtggaccag 900
gcctgggatg cacagagaat attcaaggag gccgagaagt tctttgtatc tgttggtctt 960
cctaatatga ctcaaggatt ctgggaaaat tccatgctaa cggacccagg aaatgttcag 1020
aaagcagtct gccatcccac agcttgggac ctggggaagg gcgacttcag gatccttatg 1080
tgcacaaagg tgacaatgga cgacttcctg acagctcatc atgagatggg gcatatccag 1140
tatgatatgg catatgctgc acaacctttt ctgctaagaa atggagctaa tgaaggattc 1200
catgaagctg ttggggaaat catgtcactt tctgcagcca cacctaagca tttaaaatcc 1260
attggtcttc tgtcacccga ttttcaagaa gacaatgaaa cagaaataaa cttcctgctc 1320
aaacaagcac tcacgattgt tgggactctg ccatttactt acatgttaga gaagtggagg 1380
tggatggtct ttaaagggga aattcccaaa gaccagtgga tgaaaaagtg gtgggagatg 1440
aagcgagaga tagttggggt ggtggaacct gtgccccatg atgaaacata ctgtgacccc 1500
gcatctctgt tccatgtttc taatgattac tcattcattc gatattacac aaggaccctt 1560
taccaattcc agtttcaaga agcactttgt caagcagcta aacatgaagg ccctctgcac 1620
aaatgtgaca tctcaaactc tacagaagct ggacagaaac tgttcaatat gctgaggctt 1680
ggaaaatcag aaccctggac cctagcattg gaaaatgttg taggagcaaa gaacatgaat 1740
gtaaggccac tgctcaacta ctttgagccc ttatttacct ggctgaaaga ccagaacaag 1800
aattcttttg tgggatggag taccgactgg agtccatatg cagaccatca tcaccatcac 1860
cac 1863
<210> 25
<211> 413
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 25
Met Tyr Arg Met Gln Leu Leu Ser Cys Ile Ala Leu Ser Leu Ala Leu
1 5 10 15
Val Thr Asn Ser Trp Lys Phe Met Gly Ser Lys Cys Ser Asn Ser Gly
20 25 30
Ile Glu Cys Asp Ser Ser Gly Thr Cys Ile Asn Pro Ser Asn Trp Cys
35 40 45
Asp Gly Val Ser His Cys Pro Gly Gly Glu Asp Glu Asn Arg Cys Val
50 55 60
Arg Leu Tyr Gly Pro Asn Phe Ile Leu Gln Val Tyr Ser Ser Gln Arg
65 70 75 80
Lys Ser Trp His Pro Val Cys Gln Asp Asp Trp Asn Glu Asn Tyr Gly
85 90 95
Arg Ala Ala Cys Arg Asp Met Gly Tyr Lys Asn Asn Phe Tyr Ser Ser
100 105 110
Gln Gly Ile Val Asp Asp Ser Gly Ser Thr Ser Phe Met Lys Leu Asn
115 120 125
Thr Ser Ala Gly Asn Val Asp Ile Tyr Lys Lys Leu Tyr His Ser Asp
130 135 140
Ala Cys Ser Ser Lys Ala Val Val Ser Leu Arg Cys Ile Ala Cys Gly
145 150 155 160
Val Asn Leu Asn Ser Ser Arg Gln Ser Arg Ile Val Gly Gly Glu Ser
165 170 175
Ala Leu Pro Gly Ala Trp Pro Trp Gln Val Ser Leu His Val Gln Asn
180 185 190
Val His Val Cys Gly Gly Ser Ile Ile Thr Pro Glu Trp Ile Val Thr
195 200 205
Ala Ala His Cys Val Glu Lys Pro Leu Asn Asn Pro Trp His Trp Thr
210 215 220
Ala Phe Ala Gly Ile Leu Arg Gln Ser Phe Met Phe Tyr Gly Ala Gly
225 230 235 240
Tyr Gln Val Glu Lys Val Ile Ser His Pro Asn Tyr Asp Ser Lys Thr
245 250 255
Lys Asn Asn Asp Ile Ala Leu Met Lys Leu Gln Lys Pro Leu Thr Phe
260 265 270
Asn Asp Leu Val Lys Pro Val Cys Leu Pro Asn Pro Gly Met Met Leu
275 280 285
Gln Pro Glu Gln Leu Cys Trp Ile Ser Gly Trp Gly Ala Thr Glu Glu
290 295 300
Lys Gly Lys Thr Ser Glu Val Leu Asn Ala Ala Lys Val Leu Leu Ile
305 310 315 320
Glu Thr Gln Arg Cys Asn Ser Arg Tyr Val Tyr Asp Asn Leu Ile Thr
325 330 335
Pro Ala Met Ile Cys Ala Gly Phe Leu Gln Gly Asn Val Asp Ser Cys
340 345 350
Gln Gly Asp Ser Gly Gly Pro Leu Val Thr Ser Lys Asn Asn Ile Trp
355 360 365
Trp Leu Ile Gly Asp Thr Ser Trp Gly Ser Gly Cys Ala Lys Ala Tyr
370 375 380
Arg Pro Gly Val Tyr Gly Asn Val Met Val Phe Thr Asp Trp Ile Tyr
385 390 395 400
Arg Gln Met Arg Ala Asp Gly His His His His His His
405 410
<210> 26
<211> 1242
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 26
atgtacagga tgcaactcct gtcttgcatt gcactaagtc ttgcacttgt caccaattcg 60
tggaagttta tgggttctaa atgctctaat agcgggatag aatgtgacag tagtggcact 120
tgcattaacc cttcaaactg gtgtgatggg gtaagccatt gccccggggg ggaagatgaa 180
aatagatgtg ttaggctcta cggtcccaac tttatactcc aggtatattc aagtcaacgc 240
aaatcatggc atccagtgtg tcaagacgac tggaacgaaa actatggacg cgctgcatgt 300
cgagatatgg gatataagaa taacttctat agttcacagg gaatcgtaga tgactctgga 360
tctactagtt tcatgaaact gaacacctct gccggaaacg tagatatata taaaaagctt 420
taccactccg acgcttgtag ctctaaggcc gtagttagcc tcagatgcat cgcctgcgga 480
gtaaacctca attcatctcg ccagagtagg atcgttggcg gggaaagcgc cctcccaggc 540
gcttggcctt ggcaagtttc ccttcatgtc cagaatgttc atgtatgtgg cgggtctata 600
atcaccccag aatggatcgt cacagctgcc cactgcgtgg agaaacccct caacaatcct 660
tggcattgga ccgcatttgc cggaatactg agacaatcat ttatgttcta tggagccggg 720
taccaagtcg aaaaggtcat ttcccatccc aattatgatt ccaaaaccaa aaacaatgac 780
atagccttga tgaaactcca gaagcctttg acatttaatg acctggtcaa accagtgtgc 840
ctcccaaatc ctggaatgat gttgcagcct gaacagttgt gctggatcag cggttggggt 900
gctaccgagg agaagggtaa gacaagcgag gtccttaacg ctgcaaaggt tttgctgata 960
gaaacacaga gatgtaacag ccgctatgtg tacgataacc tgatcacccc agctatgatt 1020
tgcgccgggt ttttgcaagg taacgtcgat tcttgccaag gtgactcagg cggccctctt 1080
gttacatcaa agaacaatat atggtggctt atcggcgata catcatgggg ttctggatgt 1140
gctaaagcct atcgcccagg ggtgtatggc aatgtaatgg tgtttacaga ctggatctat 1200
aggcagatgc gggctgacgg tcaccatcat caccatcact ga 1242
<210> 27
<211> 159
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 27
Met Lys Asn His Leu Leu Phe Trp Gly Val Leu Ala Val Phe Ile Lys
1 5 10 15
Ala Val His Val Lys Ala Gln Glu Asp Glu Arg Ile Val Leu Val Asp
20 25 30
Asn Lys Cys Lys Cys Ala Arg Ile Thr Ser Arg Ile Ile Arg Ser Ser
35 40 45
Glu Asp Pro Asn Glu Asp Ile Val Glu Arg Asn Ile Arg Ile Ile Val
50 55 60
Pro Leu Asn Asn Arg Glu Asn Ile Ser Asp Pro Thr Ser Pro Leu Arg
65 70 75 80
Thr Arg Phe Val Tyr His Leu Ser Asp Leu Cys Lys Lys Cys Asp Pro
85 90 95
Thr Glu Val Glu Leu Asp Asn Gln Ile Val Thr Ala Thr Gln Ser Asn
100 105 110
Ile Cys Asp Glu Asp Ser Ala Thr Glu Thr Cys Tyr Thr Tyr Asp Arg
115 120 125
Asn Lys Cys Tyr Thr Ala Val Val Pro Leu Val Tyr Gly Gly Glu Thr
130 135 140
Lys Met Val Glu Thr Ala Leu Thr Pro Asp Ala Cys Tyr Pro Asp
145 150 155
<210> 28
<211> 585
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthesized
<400> 28
Lys Ser Pro Ile Phe Gly Pro Glu Glu Val Asn Ser Val Glu Gly Asn
1 5 10 15
Ser Val Ser Ile Thr Cys Tyr Tyr Pro Pro Thr Ser Val Asn Arg His
20 25 30
Thr Arg Lys Tyr Trp Cys Arg Gln Gly Ala Arg Gly Gly Cys Ile Thr
35 40 45
Leu Ile Ser Ser Glu Gly Tyr Val Ser Ser Lys Tyr Ala Gly Arg Ala
50 55 60
Asn Leu Thr Asn Phe Pro Glu Asn Gly Thr Phe Val Val Asn Ile Ala
65 70 75 80
Gln Leu Ser Gln Asp Asp Ser Gly Arg Tyr Lys Cys Gly Leu Gly Ile
85 90 95
Asn Ser Arg Gly Leu Ser Phe Asp Val Ser Leu Glu Val Ser Gln Gly
100 105 110
Pro Gly Leu Leu Asn Asp Thr Lys Val Tyr Thr Val Asp Leu Gly Arg
115 120 125
Thr Val Thr Ile Asn Cys Pro Phe Lys Thr Glu Asn Ala Gln Lys Arg
130 135 140
Lys Ser Leu Tyr Lys Gln Ile Gly Leu Tyr Pro Val Leu Val Ile Asp
145 150 155 160
Ser Ser Gly Tyr Val Asn Pro Asn Tyr Thr Gly Arg Ile Arg Leu Asp
165 170 175
Ile Gln Gly Thr Gly Gln Leu Leu Phe Ser Val Val Ile Asn Gln Leu
180 185 190
Arg Leu Ser Asp Ala Gly Gln Tyr Leu Cys Gln Ala Gly Asp Asp Ser
195 200 205
Asn Ser Asn Lys Lys Asn Ala Asp Leu Gln Val Leu Lys Pro Glu Pro
210 215 220
Glu Leu Val Tyr Glu Asp Leu Arg Gly Ser Val Thr Phe His Cys Ala
225 230 235 240
Leu Gly Pro Glu Val Ala Asn Val Ala Lys Phe Leu Cys Arg Gln Ser
245 250 255
Ser Gly Glu Asn Cys Asp Val Val Val Asn Thr Leu Gly Lys Arg Ala
260 265 270
Pro Ala Phe Glu Gly Arg Ile Leu Leu Asn Pro Gln Asp Lys Asp Gly
275 280 285
Ser Phe Ser Val Val Ile Thr Gly Leu Arg Lys Glu Asp Ala Gly Arg
290 295 300
Tyr Leu Cys Gly Ala His Ser Asp Gly Gln Leu Gln Glu Gly Ser Pro
305 310 315 320
Ile Gln Ala Trp Gln Leu Phe Val Asn Glu Glu Ser Thr Ile Pro Arg
325 330 335
Ser Pro Thr Val Val Lys Gly Val Ala Gly Gly Ser Val Ala Val Leu
340 345 350
Cys Pro Tyr Asn Arg Lys Glu Ser Lys Ser Ile Lys Tyr Trp Cys Leu
355 360 365
Trp Glu Gly Ala Gln Asn Gly Arg Cys Pro Leu Leu Val Asp Ser Glu
370 375 380
Gly Trp Val Lys Ala Gln Tyr Glu Gly Arg Leu Ser Leu Leu Glu Glu
385 390 395 400
Pro Gly Asn Gly Thr Phe Thr Val Ile Leu Asn Gln Leu Thr Ser Arg
405 410 415
Asp Ala Gly Phe Tyr Trp Cys Leu Thr Asn Gly Asp Thr Leu Trp Arg
420 425 430
Thr Thr Val Glu Ile Lys Ile Ile Glu Gly Glu Pro Asn Leu Lys Val
435 440 445
Pro Gly Asn Val Thr Ala Val Leu Gly Glu Thr Leu Lys Val Pro Cys
450 455 460
His Phe Pro Cys Lys Phe Ser Ser Tyr Glu Lys Tyr Trp Cys Lys Trp
465 470 475 480
Asn Asn Thr Gly Cys Gln Ala Leu Pro Ser Gln Asp Glu Gly Pro Ser
485 490 495
Lys Ala Phe Val Asn Cys Asp Glu Asn Ser Arg Leu Val Ser Leu Thr
500 505 510
Leu Asn Leu Val Thr Arg Ala Asp Glu Gly Trp Tyr Trp Cys Gly Val
515 520 525
Lys Gln Gly His Phe Tyr Gly Glu Thr Ala Ala Val Tyr Val Ala Val
530 535 540
Glu Glu Arg Lys Ala Ala Gly Ser Arg Asp Val Ser Leu Ala Lys Ala
545 550 555 560
Asp Ala Ala Pro Asp Glu Lys Val Leu Asp Ser Gly Phe Arg Glu Ile
565 570 575
Glu Asn Lys Ala Ile Gln Asp Pro Arg
580 585

Claims (37)

1.一种融合蛋白，其包含与Fc结构域共价融合的变体血管紧张素转化酶2(ACE2)结构域，其中所述变体ACE2结构域相对于具有SEQ ID NO.1的全长野生型ACE2包含N-端缺失、C-端缺失或两者，其中所述变体ACE2结构域具有ACE2活性。
2.根据权利要求1所述的融合蛋白，其中所述变体ACE2结构域包含与来自全长野生型ACE2的氨基酸序列的片段具有至少98％序列同一性的氨基酸序列，其中所述片段以选自残基1-17的氨基酸残基开始，并以选自全长野生型ACE2的残基615-740的氨基酸残基结束。
3.根据权利要求1所述的融合蛋白，其中所述变体ACE2结构域包含与SEQ ID NO.3具有至少98％序列同一性的氨基酸序列。
4.根据权利要求1所述的融合蛋白，其中所述变体ACE2结构域具有比全长野生型ACE2更高的对SARS-CoV或SARS Spike蛋白的结合亲和力。
5.根据权利要求1所述的融合蛋白，其中所述Fc结构域来源于免疫球蛋白的Fc结构域，其中所述免疫球蛋白选自IgG1、IgG2、IgG3、IgG4、IgA1(d-IgA1、S-IgA1)、IgA2、IgD、IgE或IgM。
6.根据权利要求1所述的融合蛋白，其中所述Fc结构域包含Fc铰链区，并且其中所述Fc铰链区被工程改造为C220S。
7.根据权利要求1所述的融合蛋白，其中与具有SEQ ID NO.5的野生型Fc结构域相比，所述Fc结构域包含选自K322A、L234A和L235A的无效突变。
8.根据权利要求1所述的融合蛋白，其中所述Fc结构域包含与SEQ ID NO.6具有至少98％序列同一性的氨基酸序列。
9.根据权利要求1所述的融合蛋白，其包含与SEQ ID NO.7、9、11、13、15、16、17、18、19或21具有至少98％序列同一性的氨基酸序列。
10.根据权利要求1所述的融合蛋白，其中所述Fc结构域缺乏效应子功能。
11.根据权利要求1所述的融合蛋白，其中所述Fc结构域缺乏ADCC、ADCP和CDC。
12.根据权利要求1所述的融合蛋白，其分子量为约50-250kDa。
13.根据权利要求1所述的融合蛋白，其中所述Fc结构域包含IgG1 Fc结构域。
14.一种融合蛋白复合物，其包含权利要求1所述的两种融合蛋白，其中所述两种融合蛋白通过二硫键配对。
15.根据权利要求14所述的融合复合物，其中所述两种融合蛋白通过所述Fc结构域上的两个二硫键配对。
16.根据权利要求14所述的融合复合物，其中所述蛋白质复合物的分子量为约190-300kDa。
17.根据权利要求1所述的融合蛋白，其中所述融合蛋白对SARS-CoV-2、SARS-CoV或SARS刺突蛋白的结合亲和力的平衡解离常数不大于50nM。
18.一种蛋白质复合物，其包含与病毒蛋白结合的权利要求1所述的融合蛋白或权利要求14所述的融合复合物。
19.根据权利要求18所述的蛋白质复合物，其中所述病毒蛋白包含SARS-CoV-2、SARS-CoV、SARS刺突蛋白、冠状病毒、SARS病毒或其片段或组合。
20.一种分离的核酸，其编码权利要求1所述的融合蛋白。
21.一种表达载体，其包含权利要求20所述的分离的核酸。
22.一种宿主细胞，其包含权利要求20所述的核酸，其中所述宿主细胞为原核细胞或真核细胞。
23.一种产生融合蛋白的方法，其包括培养权利要求22所述的宿主细胞以产生所述融合蛋白。
24.一种蛋白质缀合物，其包含权利要求14所述的融合蛋白和药物部分，其中所述药物部分通过接头与所述融合蛋白连接，并且其中所述接头包含选自酯键、醚键、胺键、酰胺键、二硫键、酰亚胺键、砜键、磷酸酯键、磷酯键、肽键、腙键或其组合的共价键。
25.根据权利要求24所述的蛋白质缀合物，其中所述药物部分包含抗病毒剂、免疫调节试剂、显像剂或其组合。
26.根据权利要求25的蛋白质缀合物，其中所述抗病毒剂选自法匹拉韦、利巴韦林、加利地韦、瑞德西韦或其组合。
27.根据权利要求25所述的蛋白质缀合物，其中所述显像剂可以为放射性核素、荧光剂、量子点或其组合。
28.一种药物组合物，其包含权利要求14所述的融合蛋白复合物和药学上可接受的载体。
29.根据权利要求28所述的药物组合物，其还包含抗病毒剂。
30.一种药物组合物，其包含权利要求24所述的蛋白质缀合物和药学上可接受的载体。
31.一种治疗或预防受试者的病毒感染、急性呼吸窘迫综合征、肺动脉高血压或急性肺损伤的方法，其包括向所述受试者施用有效量的权利要求14所述的融合蛋白复合物。
32.根据权利要求31所述的方法，其还包括共施用有效量的治疗剂，其中所述治疗剂包含抗病毒剂。
33.根据权利要求31所述的方法，其中所述受试者是哺乳动物。
34.根据权利要求31所述的方法，其中所述病毒感染包括SARS-CoV-2、SARS-CoV、SARS刺突蛋白、冠状病毒、SARS病毒或其片段或组合的感染。
35.根据权利要求31所述的方法，其中权利要求14所述的融合蛋白复合物通过静脉内、皮下、鼻或肺施用。
36.一种包含有效浓度的权利要求14所述的融合蛋白复合物的溶液，其中所述溶液为受试者的血浆。
37.一种包含有效浓度的权利要求18所述的蛋白质复合物的溶液，其中所述溶液为受试者的血浆。
CN202180007064.2A 2021-02-10 2021-02-10 重组ace2-fc融合分子及其制备和使用方法 Pending CN115667506A (zh)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116751289A (zh) * 2023-08-03 2023-09-15 中国人民解放军军事科学院军事医学研究院 一种降低ade效应的抗西尼罗病毒中和性抗体及其应用

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2448051A1 (en) * 2001-06-04 2002-12-12 Human Genome Sciences, Inc. Methods and compositions for modulating ace-2 activity
AU2006236225C1 (en) * 2005-04-19 2013-05-02 Seagen Inc. Humanized anti-CD70 binding agents and uses thereof
WO2008060331A2 (en) * 2006-05-19 2008-05-22 Amgen Inc. Antibodies to sars coronavirus
JP7332157B2 (ja) * 2017-01-24 2023-08-23 ノースウェスタン ユニバーシティ アンジオテンシン変換酵素２（ａｃｅ２）の活性な低分子量変異体
CN112226424A (zh) * 2020-09-27 2021-01-15 苏州新格诺康生物技术有限公司 一种用于治疗COVID-19的ACE2-Fc融合蛋白功能测试方法

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116751289A (zh) * 2023-08-03 2023-09-15 中国人民解放军军事科学院军事医学研究院 一种降低ade效应的抗西尼罗病毒中和性抗体及其应用
CN116751289B (zh) * 2023-08-03 2023-10-20 中国人民解放军军事科学院军事医学研究院 一种降低ade效应的抗西尼罗病毒中和性抗体及其应用

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