CN113667016A - 一种构建冠状病毒抗体的平台 - Google Patents

一种构建冠状病毒抗体的平台 Download PDF

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CN113667016A
CN113667016A CN202010414651.0A CN202010414651A CN113667016A CN 113667016 A CN113667016 A CN 113667016A CN 202010414651 A CN202010414651 A CN 202010414651A CN 113667016 A CN113667016 A CN 113667016A
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缪小牛
许英达
罗羿
袁志军
陈连娣
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Biotheus Inc
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Abstract

本发明涉及一种使用抗SARS‑CoV2 Spike 1的抗体和细胞受体ACE2胞外区的融合蛋白用于治疗SARS‑CoV2感染的方法。具体地，本发明提供了一种利用该方法开发的抗S1的抗体和受体ACE2的融合蛋白。该融合蛋白可以和S1以高亲和力结合，并可以阻断S1蛋白和ACE2的相互作用，中和病毒的细胞侵染活性。并且，本发明提供了该融合蛋白的表达载体和能够表达该融合蛋白的宿主细胞，以及本发明抗体的生产方法。

Description

一种构建冠状病毒抗体的平台
技术领域
本发明属于生物技术领域，具体涉及一种构建冠状病毒抗体的平台。
背景技术
在急性传染病中，绝大部分都是病毒性传染病，病毒性传染病的发病率高，死亡率也很高。由于检测和诊断手段有限，导致新病毒引发的新疫情爆发往往具有突发性、随机性和不可预测性等特点，一旦爆发，如无有效的防治手段，极易造成大规模流行，严重威胁人民健康生命安全。
2019-nCoV冠状病毒属于冠状病毒科冠状病毒属，为具有包膜的单链正义RNA病毒。和其他已知冠状病毒类似，2019-nCoV冠状病毒也经过吸附、穿入、脱壳、生物合成、子代病毒的组装与释放等几个过程完成子代病毒的增殖。2019-nCoV冠状病毒感染宿主细胞起始于病毒包膜表面的刺突糖蛋白与宿主细胞表面的人血管紧张素转化酶2(ACE2)结合，随后发生膜融合，病毒进入宿主细胞，在细胞溶酶体等细胞器作用下，释放出病毒的遗传物质单链正义RNA，在宿主细胞的线粒体、核糖体等蛋白质合成元件以及必须的原料等作用下，翻译产生多聚蛋白。之后，2019-nCoV冠状病毒的两大必需半胱氨酸蛋白酶：木瓜样蛋白酶(papain-like protease,PL pro)和3C样蛋白酶(3C-likeprotease,3CLpro)在特定位点切割加工多聚蛋白前体，产生多个对病毒生命周期非常重要的非结构蛋白。在这些非结构蛋白的作用下，病毒RNA复制出子代病毒核酸物质，并大量翻译出所需的结构蛋白，完成子代病毒的组装和释放。2019-nCoV冠状病毒感染细胞的生命周期的任何环节或关键酶均可以作为抗病毒药物的研究靶点，如水解切割多聚蛋白前体的半胱氨酸蛋白酶PLpro和3CLpro，负责完成子代病毒遗传物质复制的RNA聚合酶等。
迄今为止，2019-nCoV病毒的传播途径未完全掌握，已知能通过飞沫和接触传播，且存在人传人、医务人员感染，一定社区传播风险，且病毒存在变异的可能。目前对于新型冠状病毒所致疾病没有特异的预防和治疗方法。
目前，针对SARS-CoV-2(同2019-nCoV)冠状病毒导致的严重肺炎疾病尚无特效的疫苗和抗病毒药物。这些感染性疾病严重影响了人类的生命健康，研发效果好的抗病毒药物迫在眉睫。针对SARS-CoV-2冠状病毒开发出低毒高效的抗病毒药物，以满足国内外SARS-CoV-2冠状病毒感染患者的临床需求，具有重大的社会意义。
综上所述，本领域迫切需要开发针对SARS-CoV-2冠状病毒有效诊断和治疗方法，以用于诊断和治疗新型冠状病毒感染引起的肺炎。
发明内容
本发明的目的就是提供一种高效的针对SARS-CoV-2冠状病毒有效诊断和治疗方法。
本发明的另一目的是提供一种能够改进抗SARS-Cov2或其他病毒抗体中和能力的技术手段。
本发明的另一目的是提供一种能够抗SARS-Cov2或其他病毒的抗体融合蛋白技术手段。
在本发明的第一方面，提供了一种改进抗冠状病毒抗体中和能力的方法，包括步骤：将所述待改进的抗冠状病毒抗体与人血管紧张素转化酶2(ACE2)或其片段进行融合，从而得到一融合蛋白；
所述的融合蛋白与单独的所述抗冠状病毒抗体相比，抗原中和能力提高≥10倍(较佳地，提高≥20倍)。
在另一优选例中，所述的冠状病毒是SAR-CoV-2病毒。
在另一优选例中，所述的抗体特异性地结合SARS-CoV-2Spike 1蛋白。
在另一优选例中，所述的人ACE2或其片段的氨基酸序列如SEQ ID NO:1、2、3或4所示；或在SEQ ID NO:1、2、3或4所示序列的基础上，经过添加、缺失、修饰和/或取代至少一个氨基酸的，并能够保留SARS-CoV-2Spike 1蛋白结合亲和力的衍生序列。
在另一优选例中，所述的经过添加、缺失、修饰和/或取代至少一个氨基酸的，并能够保留SARS-CoV-2Spike 1结合亲和力的衍生序列为同源性或序列相同性为至少85％、至少90％、至少95％、至少96％、至少97％、至少98％或至少99％的氨基酸序列。
在另一优选例中，所述的人ACE2或其片段是人ACE2的胞外区全长蛋白。
在另一优选例中，所述的人ACE2或其片段的氨基酸序列如SEQ ID NO:4所示。
在另一优选例中，所述的融合蛋白中，所述的人ACE2或其片段被融合至所述抗体的重链的N端、重链的C端、轻链的N端，和/或轻链的C端。
在另一优选例中，所述的融合蛋白具有相互堆成的两对肽链，每对肽链含有重链H链和轻链L链，所有的肽链均由二硫键相连，其中任意一对肽链从C端到C端具有如式I至V所示的H链和L链的结构，
Figure BDA0002494502590000031
其中，
L1、L2、L3、L4、L5和L6均为连接肽序列，其序列各自独立地为(G4S)n或其变体，其中，n为正整数(例如1、2、3、4、5或6)，优选地，n＝4；
ACE2代表人ACE2或其片段；
VH代表抗SARS-Cov2 Spike 1蛋白抗体的重链可变区；
VL代表抗SARS-Cov2 Spike 1蛋白抗体的轻链可变区；
CH1、CH2和CH3分别代表抗SARS-Cov2 Spike 1蛋白抗体的重链恒定区CH1、CH2和CH3；
CL代表抗SARS-Cov2 Spike 1蛋白抗体的轻链恒定区；
“～”代表二硫键；
“-”代表肽键。
在另一优选例中，所述(G4S)n的变体包括：将所述序列中性能相近或相似的氨基酸进行取代所获得的(G4S)n接头序列的变体，如将一个或多个S分别突变为T；或者在所述序列中插入1-3个氨基酸。
在另一优选例中，所述连接肽序列的氨基酸序列如SEQ ID NO:10所示。
在另一优选例中，所述的方法是非治疗性，并且是非诊断性的。
在另一优选例中，所述抗体的重链恒定区CH1、CH2和CH3、以及轻链恒定区CL均来源于人IgG1或IgG4，优选为人IgG1。
在另一优选例中，所述式I至式V中的VH-CH1-CH2-CH3区段(即待改进的89C8抗体的重链)的氨基酸序列如SEQ ID NO:11所示，或与SEQ ID NO:11的序列具有≥85％(优选地90％，更优选地95％)的序列同一性。
在另一优选例中，所述式I至式V中的VL-CL区段(即待改进的89C8抗体的轻链)的氨基酸序列如SEQ ID NO:16所示，或与SEQ ID NO:16的序列具有≥85％(优选地90％，更优选地95％)的序列同一性。
在另一优选例中，所述融合蛋白中的H链具有如SEQ ID NO:15所示的氨基酸序列，并且融合蛋白中的L链具有如SEQ ID NO:16所示的氨基酸序列；
或者，所述融合蛋白中的H链具有如SEQ ID NO:15所示的氨基酸序列，并且融合蛋白中的L链具有如SEQ ID NO:20所示的氨基酸序列；
或者，所述融合蛋白中的H链具有如SEQ ID NO:11所示的氨基酸序列，并且融合蛋白中的L链具有如SEQ ID NO:20所示的氨基酸序列。
在另一优选例中，所述式I至式V中的VH-CH1-CH2-CH3区段(即待改进的CR3022抗体的重链)的氨基酸序列如SEQ ID NO:21所示，或与SEQ ID NO:21的序列具有≥85％(优选地90％，更优选地95％)的序列同一性。
在另一优选例中，所述式I至式V中的VL-CL区段(即待改进的CR3022抗体的轻链)的氨基酸序列如SEQ ID NO:22所示，或与SEQ ID NO:22的序列具有≥85％(优选地90％，更优选地95％)的序列同一性。
在另一优选例中，所述融合蛋白中的H链具有如SEQ ID NO:26所示的氨基酸序列，并且融合蛋白中的L链具有如SEQ ID NO:22所示的氨基酸序列；
或者，所述融合蛋白中的H链具有如SEQ ID NO:26所示的氨基酸序列，并且融合蛋白中的L链具有如SEQ ID NO:30所示的氨基酸序列；
或者，所述融合蛋白中的H链具有如SEQ ID NO:21所示的氨基酸序列，并且融合蛋白中的L链具有如SEQ ID NO:30所示的氨基酸序列。
在本发明的第二方面，提供了一种融合蛋白，所述的融合蛋白的序列中包含：
(a)抗SARS-Cov2 Spike 1蛋白抗体的序列；和
(b)人ACE2或其片段的序列。
在另一优选例中，所述的融合蛋白中，所述的人ACE2或其片段位于所述SARS-Cov2Spike 1蛋白抗体的重链的N端、重链的C端、轻链的N端，和/或轻链的C端。
在本发明的第三方面，提供了一种分离的多核苷酸，所述多核苷酸编码如本发明第二方面所述的融合蛋白。
在另一优选例中，所述的多核苷酸中，编码H链的多核苷酸和编码L链的多核苷酸的比例为1:2至3:1，优选地为2:1。
在本发明的第四方面，提供了一种载体，所述载体含有如本发明第三方面所述的多核苷酸。
在另一优选例中，所述载体同时含有如本发明第三方面所述多核苷酸中的所有多核苷酸。
在另一优选例中，所述载体分别含有如本发明第三方面所述的编码H链的多核苷酸和编码L链的多核苷酸。
在另一优选例中，所述载体为表达载体。
在另一优选例中，所述载体包括质粒、噬菌体、酵母质粒、植物细胞病毒、哺乳动物细胞病毒如腺病毒、逆转录病毒、或其他载体。
在本发明的第五方面，提供了一种遗传工程化的宿主细胞，所述宿主细胞含有如本发明第四方面所述的载体或基因组中整合有如本发明第三方面所述的多核苷酸。
在本发明的第六方面，提供了一种抗体偶联物，所述抗体偶联物含有：
(a)靶向部分，所述靶向部分包括如本发明第二方面所述的融合蛋白；和
(b)与所述靶向部分偶联的偶联部分，所述偶联部分选自下组：可检测标记物、药物、毒素、细胞因子、放射性核素、酶、金纳米颗粒/纳米棒、纳米磁粒、病毒外壳蛋白或VLP，或其组合。
在另一优选例中，所述的靶向部分与所述的偶联部分通过化学键或接头进行偶联。
在另一优选例中，所述的放射性核素包括：
(i)诊断用同位素，所述的诊断用同位素选自下组：Tc-99m、Ga-68、F-18、I-123、I-125、I-131、In-111、Ga-67、Cu-64、Zr-89、C-11、Lu-177、Re-188、或其组合；和/或
(ii)治疗用同位素，所述的治疗用同位素选自下组：Lu-177、Y-90、Ac-225、As-211、Bi-212、Bi-213、Cs-137、Cr-51、Co-60、Dy-165、Er-169、Fm-255、Au-198、Ho-166、I-125、I-131、Ir-192、Fe-59、Pb-212、Mo-99、Pd-103、P-32、K-42、Re-186、Re-188、Sm-153、Ra223、Ru-106、Na24、Sr89、Tb-149、Th-227、Xe-133Yb-169、Yb-177、或其组合。
在另一优选例中，所述偶联部分为药物或毒素。
在另一优选例中，所述的药物为细胞毒性药物。
在另一优选例中，所述的细胞毒性药物选自下组：抗微管蛋白药物、DNA小沟结合试剂、DNA复制抑制剂、烷化试剂、抗生素、叶酸拮抗物、抗代谢药物、化疗增敏剂、拓扑异构酶抑制剂、长春花生物碱、或其组合。
特别有用的细胞毒性药物类的例子包括，例如，DNA小沟结合试剂、DNA烷基化试剂、和微管蛋白抑制剂、典型的细胞毒性药物包括、例如奥瑞他汀(auristatins)、喜树碱(camptothecins)、多卡霉素/倍癌霉素(duocarmycins)、依托泊甙(etoposides)、美登木素(maytansines)和美登素类化合物(maytansinoids)(例如DM1和DM4)、紫杉烷(taxanes)、苯二氮卓类(benzodiazepines)或者含有苯二氮卓的药物(benzodiazepine containingdrugs)(例如吡咯并[1,4]苯二氮卓类(PBDs)，吲哚啉苯并二氮卓类(indolinobenzodiazepines)和噁唑烷并苯并二氮卓类(oxazolidinobenzodiazepines))、长春花生物碱(vinca alkaloids)、或其组合。
在另一优选例中，所述的毒素选自下组：
耳他汀类(例如，耳他汀E、耳他汀F、MMAE和MMAF)、金霉素、类美坦西醇、篦麻毒素、篦麻毒素A-链、考布他汀、多卡米星、多拉司他汀、阿霉素、柔红霉素、紫杉醇、顺铂、cc1065、溴化乙锭、丝裂霉素、依托泊甙、替诺泊甙(tenoposide)、长春新碱、长春碱、秋水仙素、二羟基炭疽菌素二酮、放线菌素、白喉毒素、假单胞菌外毒素(PE)A、PE40、相思豆毒素、相思豆毒素A链、蒴莲根毒素A链、α-八叠球菌、白树毒素、迈托毒素(mitogellin)、局限曲菌素(retstrictocin)、酚霉素、依诺霉素、麻疯树毒蛋白(curicin)、巴豆毒素、卡奇霉素、肥皂草(Sapaonaria officinalis)抑制剂、糖皮质激素、或其组合。
在另一优选例中，所述偶联部分为可检测标记物。
在另一优选例中，所述偶联物选自：荧光或发光标记物、放射性标记物、MRI(磁共振成像)或CT(电子计算机X射线断层扫描技术)造影剂、或能够产生可检测产物的酶、放射性核素、生物毒素、细胞因子(如IL-2)、抗体、抗体Fc片段、抗体scFv片段、金纳米颗粒/纳米棒、病毒颗粒、脂质体、纳米磁粒、前药激活酶(如DT-心肌黄酶(DTD)或联苯基水解酶-样蛋白质(BPHL))、化疗剂(如顺铂)。
在另一优选例中，所述免疫偶联物含有：多价(如二价)的(a)。
在另一优选例中，所述多价是指在所述免疫偶联物的氨基酸序列中包含多个重复的(a)。
在本发明的第七方面，提供了一种免疫细胞，所述免疫细胞表达或在细胞膜外暴露有如本发明第二方面所述的融合蛋白。
在另一优选例中，所述的免疫细胞包括NK细胞、T细胞。
在另一优选例中，所述的免疫细胞来自人或非人哺乳动物(如鼠)。
在本发明的第八方面，提供了一种药物组合物，所述药物组合物含有：
(i)活性成分，所述活性成分选自下组：如本发明第二方面所述的融合蛋白、如本发明第六方面所述的抗体偶联物、如本发明第七方面所述的免疫细胞，或其组合；以及
(ii)药学上可接受的载体。
在另一优选例中，所述的药物组合物为液态制剂。
在另一优选例中，所述的药物组合物为注射剂。
在另一优选例中，所述的药物组合物包括0.01～99.99％的如本发明第二方面所述的融合蛋白、如本发明第六方面所述的抗体偶联物、如本发明第七方面所述的免疫细胞，或其组合，以及0.01～99.99％的药用载体，所述百分比为占所述药物组合物的质量百分比。
在本发明的第九方面，提供了一种活性成分的用途，被用于(a)制备SARS-CoV2病毒感染的诊断试剂或试剂盒；和/或(b)制备预防和/或治疗SARS-CoV2病毒感染的药物；
其中，所述活性成分选自下组：如本发明第二方面所述的融合蛋白、如本发明第六方面所述的抗体偶联物、如本发明第七方面所述的免疫细胞，或其组合。
在另一优选例中，所述的诊断试剂为检测片或检测板。
在另一优选例中，所述诊断试剂或试剂盒用于：检测样品中SARS-CoV-2Spike1蛋白。
在本发明的第十方面，提供了一种制备如本发明第二方面所述的融合蛋白的方法，包括步骤：
(i)在适合表达的条件下，培养如本发明第五方面所述的宿主细胞；和
(ii)从培养物中分离出所述的重组多肽。
在本发明的第十一方面，提供了一种体外检测样品中SARS-CoV-2病毒或SARS-CoV-2Spike 1蛋白的方法，所述方法包括步骤：
(1)在体外，将所述样品与如本发明第二方面所述的融合蛋白接触；
(2)检测是否形成抗原-抗体复合物，其中形成复合物就表示样品中存在SARS-CoV-2病毒或SARS-CoV-2Spike 1蛋白。
在另一优选例中，所述的检测包括诊断性的或非诊断性的。
在本发明的第十二方面，提供了一种试剂盒，所述试剂盒中包括：
(1)第一容器，所述第一容器中含有如本发明第二方面所述的融合蛋白；和/或
(2)第二容器，所述第二容器中含有抗如本发明第二方面所述融合蛋白的二抗；
或者，
所述试剂盒含有一检测板，所述检测板包括：基片(支撑板)和测试条，所述的测试条含有如本发明第五方面所述的融合蛋白、如本发明第六方面所述的抗体偶联物、如本发明第七方面所述的免疫细胞，或其组合。
在本发明的第十三方面，提供了一种治疗有所需要的对象的病毒感染类疾病的方法，包括步骤：
(a)制备一融合蛋白，所述融合蛋白的氨基酸序列中包含：抗病毒表面蛋白的抗体，以及所述对象体内介导所述病毒侵染的蛋白或其片段。
在另一优选例中，所述的病毒包括：SARS、MERS，或其他病毒。
在另一优选例中，所述的介导病毒侵染的蛋白例如SARS所对应的ACE2，或MERS所对应的DPP2等。
在本发明的第十四方面，提供了一种抗冠状病毒的融合蛋白的制备方法，包括步骤：将一抗冠状病毒抗体与人血管紧张素转化酶2(ACE2)或其片段进行融合，从而得到所述融合蛋白。
应理解，在本发明范围内中，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。限于篇幅，在此不再一一累述。
附图说明
图1显示了本发明融合蛋白的5种基本结构。
图2显示了89C8-ACE2-F1融合蛋白和89C8抗体分别与S1蛋白的亲和力。
图3显示了89C8-ACE2-F1融合蛋白与293-Spike细胞的结合活性。
图4显示了89C8-ACE2-F1融合蛋白对ACE2与Spike 1蛋白结合的阻断活性。
图5显示了89C8-ACE2-F1融合蛋白、89C8和ACE2-Fc对假病毒的侵染活性的中和能力。
具体实施方式
本发明人经过广泛而深入的研究，经过大量的筛选，首次开发了一种改进抗SARS-CoV2 S1抗体的中和能力的方法。研究表明，将ACE2胞外区与所述抗体进行融合后，所得的融合蛋白与SARS-CoV2 S1蛋白的亲和力显著提高，并且所述融合蛋白对病毒的中和活性显著增强。
在此基础上，完成了本发明。
本发明的改进方法(或平台)
在本发明中，提供了一种改进抗冠状病毒抗体中和能力的方法，包括步骤：将所述待改进的抗冠状病毒抗体与人血管紧张素转化酶2(ACE2)或其片段进行融合，从而得到一融合蛋白；所述的融合蛋白与单独的所述抗冠状病毒抗体相比，抗原中和能力提高≥10倍(较佳地，提高≥20倍)。
优选地，本发明所述的冠状病毒是SAR-CoV-2病毒。更加优选地，所述的抗体特异性地结合SARS-CoV-2Spike 1蛋白。
在一个实施方式中，所述的人ACE2或其片段的氨基酸序列如SEQ ID NO:1、2、3或4所示；或在SEQ ID NO:1、2、3或4所示序列的基础上，经过添加、缺失、修饰和/或取代至少一个氨基酸的，并能够保留SARS-CoV-2Spike 1蛋白结合亲和力的衍生序列。特别优选地，所述的人ACE2或其片段是指ACE2胞外区的全长蛋白，其氨基酸序列如SEQ ID NO:4所示。
优选地，所述的融合蛋白中，所述的人ACE2或其片段被融合至所述抗体的重链的N端、重链的C端、轻链的N端，和/或轻链的C端。
在所述的融合蛋白中，待改进的抗体可以是与SARS-CoV2 S1蛋白具有高亲和力的任何抗体。
在一个实施方式中，所述的抗体是89C8，其重链的氨基酸序列如SEQ ID NO:11所示，并且其轻链的氨基酸序列如SEQ ID NO:16所示。
当所述的抗体为89C8时，所述的融合蛋白中具有如SEQ ID NO:15所示的氨基酸序列的H链，以及如SEQ ID NO:16所示的氨基酸序列的L链(89C8-ACE-F1)；
或者，所述的融合蛋白具有如SEQ ID NO:15所示的氨基酸序列的H链，以及如SEQID NO:20所示的氨基酸序列的L链(89C8-ACE-F2)；
或者，所述的融合蛋白具有如SEQ ID NO:11所示的氨基酸序列的H链，以及如SEQID NO:20所示的氨基酸序列的L链(89C8-ACE-F3)。
在一个实施方式中，所述的抗体是CR3022，其重链的氨基酸序列如SEQ ID NO:21所示，并且其轻链的氨基酸序列如SEQ ID NO:22所示。
当所述的抗体为CR3022时，所述的融合蛋白中具有如SEQ ID NO:26所示的氨基酸序列的H链，以及如SEQ ID NO:22所示的氨基酸序列的L链(CR3022-ACE-F1)；
或者，所述的融合蛋白具有如SEQ ID NO:26所示的氨基酸序列的H链，以及如SEQID NO:30所示的氨基酸序列的L链(CR3022-ACE-F2)；
或者，所述的融合蛋白具有如SEQ ID NO:21所示的氨基酸序列的H链，以及如SEQID NO:30所示的氨基酸序列的L链(CR3022-ACE-F3)。
应用
本发明还提供了本发明融合蛋白、抗体偶联物ADC、重组蛋白、和/或免疫细胞的用途，例如用于制备诊断制剂或制备药物。
较佳地，所述的药物是用于预防和/或治疗SARS-CoV-2病毒感染的疾病的药物。
检测用途和试剂盒
本发明的融合蛋白或其ADC可用于检测应用，例如用于检测样本，从而提供诊断信息。
本发明中，所采用的样本(样品)包括细胞、组织样本和活检标本。
较佳地，所述的样本是来自于检测对象的血液样本。
本发明使用的术语“活检”应包括本领域技术人员已知的所有种类的活检。因此本发明中使用的活检可以包括例如肿瘤的切除样本、通过内窥镜方法或器官的穿刺或针刺活检制备的组织样本。
本发明中使用的样本包括固定的或保存的细胞或组织样本。
本发明还提供了一种指含有本发明的融合蛋白(或其片段)的试剂盒，在本发明的一个优选例中，所述的试剂盒还包括容器、使用说明书、缓冲剂等。在优选例中，本发明的融合蛋白可以固定于检测板。
药物组合物
本发明还提供了一种组合物。在优选例中，所述的组合物是药物组合物，它含有上述的融合蛋白或其活性片段或其融合蛋白或其ADC或相应的免疫细胞，以及药学上可接受的载体。通常，可将这些物质配制于无毒的、惰性的和药学上可接受的水性载体介质中，其中pH通常约为5-8，较佳地pH约为6-8，尽管pH值可随被配制物质的性质以及待治疗的病症而有所变化。
配制好的药物组合物可以通过常规途径进行给药，其中包括(但并不限于)：瘤内、腹膜内、静脉内、或局部给药。典型地，本发明所述的药物组合物的给药途径较佳地为注射给药或口服给药。所述注射给药较佳地包括静脉注射、肌肉注射、腹腔注射、皮内注射或皮下注射等途径。所述的药物组合物为本领域常规的各种剂型，较佳地为固体、半固体或液体的形式，可以为水溶液、非水溶液或混悬液，更佳地为片剂、胶囊、颗粒剂、注射剂或输注剂等。
本发明所述融合蛋白也可以是由核苷酸序列在细胞内表达用于的细胞治疗，比如，所述融合蛋白用于嵌合抗原受体T细胞免疫疗法(CAR-T)等。
本发明所述的药物组合物是用于预防和/或治疗SARS-CoV-2病毒感染的疾病的药物组合物。
本发明的药物组合物可直接用于结合SARS-CoV-2Spike 1蛋白分子，因而可用于预防和治疗病毒感染所引发的疾病。
本发明的药物组合物含有安全有效量(如0.001-99wt％,较佳地0.01-90wt％，更佳地0.1-80wt％)的本发明上述的融合蛋白(或其偶联物)以及药学上可接受的载体或赋形剂。这类载体包括(但并不限于)：盐水、缓冲液、葡萄糖、水、甘油、乙醇、及其组合。药物制剂应与给药方式相匹配。本发明的药物组合物可以被制成针剂形式，例如用生理盐水或含有葡萄糖和其他辅剂的水溶液通过常规方法进行制备。药物组合物如针剂、溶液宜在无菌条件下制造。活性成分的给药量是治疗有效量，例如每天约1微克/千克体重-约5毫克/千克体重。此外，本发明的融合蛋白还可与其他治疗剂一起使用。
本发明中，较佳地，本发明所述的药物组合物还包括一种或多种药用载体。所述的药用载体为本领域常规药用载体，所述的药用载体可以为任意合适的生理学或药学上可接受的药物辅料。所述的药物辅料为本领域常规的药物辅料，较佳的包括药学上可接受的赋形剂、填充剂或稀释剂等。更佳地，所述的药物组合物包括0.01～99.99％的上述蛋白质和0.01～99.99％的药用载体，所述百分比为占所述药物组合物的质量百分比。
本发明中，较佳地，所述的药物组合物的施用量为有效量，所述有效量为能够缓解或延迟疾病、退化性或损伤性病症进展的量。所述有效量可以以个体基础来测定，并将部分基于待治疗症状和所寻求结果的考虑。本领域技术人员可以通过使用个体基础等上述因素和使用不超过常规的实验来确定有效量。
使用药物组合物时，是将安全有效量的免疫偶联物施用于哺乳动物，其中该安全有效量通常至少约10微克/千克体重，而且在大多数情况下不超过约50毫克/千克体重，较佳地该剂量是约10微克/千克体重-约20毫克/千克体重。当然，具体剂量还应考虑给药途径、病人健康状况等因素，这些都是熟练医师技能范围之内的。
本发明提供上述药物组合物在制备预防和/或治疗SARS-CoV-2病毒感染的疾病的药物中的应用。较佳地，所述SARS-CoV-2病毒感染的疾病为肺炎。
冠状病毒
如本文所用，术语“新型冠状病毒”、“2019-nCov”或“SARS-CoV-2”可互换使用，该2019新型冠状病毒是已知感染人的第7种冠状病毒，并且造成新冠肺炎(COVID-19)，是威胁全球人类健康的严重传染性疾病之一。
冠状病毒(Coronavirus，CoV)属于套式病毒目(Nidovirales)冠状病毒科(Coronaviridae)，是一种有包膜的正链RNA病毒，其亚科包含α、β、δ及γ四属。
目前已知的感染人的冠状病毒中，HCoV-229E和HCoV-NL63属于α属冠状病毒，HCoV-OC43、SARS-CoV、HCoV-HKU1、MERS-CoV和SARS-CoV-2均为β属冠状病毒。SARS-CoV-2也被称为2019-nCov。
2003年和2012年分别爆发的高致病性冠状病毒“非典”SARS-CoV和“中东呼吸综合征”MERS-CoV均属于β属冠状病毒。2019年年底爆发的新型冠状病毒(SARS-CoV-2)与SARS-CoV有约80％相似性、与MERS-CoV有40％的相似性，也属于β属冠状病毒。
该类病毒的基因组是一条单股正链RNA，是基因组最大的RNA病毒之一，编码包括复制酶、刺突蛋白、囊膜蛋白、包膜蛋白和核壳蛋白等。在病毒复制的初始阶段，基因组被翻译成两条长达几千个氨基酸的肽链即前体多聚蛋白(Polyprotein)，随后前体蛋白被蛋白酶切割生成非结构蛋白(如RNA聚合酶和解旋酶)和结构蛋白(如刺突蛋白)及辅助蛋白。
本发明的主要优点包括：
1)本发明融合蛋白可以和S1蛋白以高亲和力结合。
2)本发明可以阻断S1蛋白和ACE2的相互作用，阻断活性比ACE2蛋白明显提高。
3)本发明可以中和Spike1假病毒的细胞侵染活性，中和活性比单抗明显提高。
4)本发明的方法可以用于SARS-CoV2及其他病毒感染(如SARS、MERS等)类疾病的治疗。
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件，例如Sambrook等人，分子克隆：实验室手册(New York:Cold Spring HarborLaboratory Press,1989)中所述的条件，或按照制造厂商所建议的条件。除非另外说明，否则百分比和份数是重量百分比和重量份数。
本发明融合蛋白名称以及本发明的序列
在本发明中，涉及到多个融合蛋白,其名称以及对应的重链和轻链的名称总结于表1中。
表1融合蛋白列表
Figure BDA0002494502590000141
Figure BDA0002494502590000151
表2本发明涉及的序列列表
Figure BDA0002494502590000152
Figure BDA0002494502590000161
Figure BDA0002494502590000171
Figure BDA0002494502590000181
Figure BDA0002494502590000191
实施例1：IgG-ACE2融合蛋白的克隆和表达
研究表明，ACE2和CAR-CoV-2S1蛋白的结合位点集中在其N端的23个氨基酸中。
在本实施例中，使用了含有23个氨基酸、45个氨基酸、98个氨基酸的ACE2截短蛋白以及ACE2胞外区全长蛋白作为融合蛋白中靶向及中和作用的部分，将S1抗体作为融合蛋白的靶向部分，形成S1抗体/ACE2胞外区融合蛋白。融合蛋白的结构如图2所示。ACE2截短及全长蛋白的氨基酸序列如下：
23个氨基酸的ACE2截短蛋白序列(SEQ ID NO:1)：
EQAKTFLDKFNHEAEDLFYQS
45个氨基酸的ACE2截短蛋白序列(SEQ ID NO:2)：
QSTIEEQAKTFLDKFNHEAEDLFYQSSLASWNYNTNITEENVQNM
98个氨基酸的ACE2截短蛋白序列(SEQ ID NO:3)：
QSTIEEQAKTFLDKFNHEAEDLFYQSSLASWNYNTNITEENVQNMNNAGDKWSAFLKEQSTLAQMYPLQEIQNLTVKLQLQALQQNGSSVLSEDKSKR
ACE2胞外区全长序列(SEQ ID NO:4)：
QSTIEEQAKTFLDKFNHEAEDLFYQSSLASWNYNTNITEENVQNMNNAGDKWSAFLKEQSTLAQMYPLQEIQNLTVKLQLQALQQNGSSVLSEDKSKRLNTILNTMSTIYSTGKVCNPDNPQECLLLEPGLNEIMANSLDYNERLWAWESWRSEVGKQLRPLYEEYVVLKNEMARANHYEDYGDYWRGDYEVNGVDGYDYSRGQLIEDVEHTFEEIKPLYEHLHAYVRAKLMNAYPSYISPIGCLPAHLLGDMWGRFWTNLYSLTVPFGQKPNIDVTDAMVDQAWDAQRIFKEAEKFFVSVGLPNMTQGFWENSMLTDPGNVQKAVCHPTAWDLGKGDFRILMCTKVTMDDFLTAHHEMGHIQYDMAYAAQPFLLRNGANEGFHEAVGEIMSLSAATPKHLKSIGLLSPDFQEDNETEINFLLKQALTIVGTLPFTYMLEKWRWMVFKGEIPKDQWMKKWWEMKREIVGVVEPVPHDETYCDPASLFHVSNDYSFIRYYTRTLYQFQFQEALCQAAKHEGPLHKCDISNSTEAGQKLFNMLRLGKSEPWTLALENVVGAKNMNVRPLLNYFEPLFTWLKDQNKNSFVGWSTDWSPYADQSIKVRISLKSALGDKAYEWNDNEMYLFRSSVAYAMRQYFLKVKNQMILFGEEDVRVANLKPRISFNFFVTAPKNVSDIIPRTEVEKAIRMSR
利用分子克隆技术将本发明ACE2的截短蛋白或者全长蛋白通过(G4S)4G连接到抗S1抗体89C8或已报道的抗SARS S1蛋白抗体CR3022(专利号：US2008/0014204A1)的重链C端(结构1，F1)、轻链N端(结构3，F3)、同时连到重链C端和轻链N端(结构2，F2)、重链N端(结构4，F4)及轻链C端(结构5，F5)。
随后，将所得的各融合蛋白的基因序列利用同源重组酶(购自Vazyme)和EcoR I/Not I双酶切线性化的pCDNA3.1载体中，流程按照商品说明书。同源重组产物化转入Top10感受态细胞，涂布氨苄抗性平板，37℃培养过夜，挑取单克隆测序。
根据所需转染体积传代HEK293细胞，转染前一天将细胞密度调整至1.2×106个细胞/ml。取适量的MEM(购自Gibco)培养基作为转染缓冲液，加入重组基因质粒，质粒配对如表1所示，混匀，过滤除菌。按PEI(购自Polysciences)：质粒＝3:1比例加入PEI，混匀后室温孵育20min，倒入HEK293细胞中，36.5℃，8％CO2培养，等细胞活力降低到60％以下离心收集上清利用蛋白A磁珠(购自金斯瑞)分选法纯化目的蛋白。将磁珠用适当体积的结合缓冲液(PBS+0.1％吐温20，pH 7.4)重悬(1-4倍磁珠体积)后加入至待纯化样品中，室温孵育1h，期间温柔振荡。样品置于磁力架上(购自海狸)，弃去上清，磁珠用结合缓冲液清洗3遍。按照磁珠体积的3-5倍体积加入洗脱缓冲液(0.1M柠檬酸钠，pH3.2)室温振荡5-10min，置回磁力架上，收集洗脱缓冲液，转移至已加入中和缓冲液(1M Tris，pH 8.54)的收集管中混匀。
从而得到结构如图1所示的融合蛋白。
实施例2：S1抗体/ACE2胞外区融合蛋白和S1蛋白的结合活性
AHC传感器在分析缓冲液中线下平衡30min，然后线上检测60s建立基线，在线加载如上所述获得的经纯化的S1抗体/ACE2胞外区(截短)融合蛋白至AHC传感器上。再将传感器放入100nM的S1-His抗原中作用240s，之后将传感器转移至PBS中解离1000s。使用1:1结合模型进行动力学的分析。
实验结果如下表所示，使用截短的ACE2构建的抗体融合蛋白和S1的亲和力与抗体89C8相似，没有产生加和作用。
表2 S1抗体/ACE2胞外区(截短)融合蛋白候选分子亲和力
编号 K<sub>D</sub>(M) kon(1/Ms) kdis(1/s)
89C8-23-F1 1.10E-08 3.93E+04 4.32E-04
89C8-45-F1 8.50E-09 4.65E+04 3.93E-04
89C8-98-F1 1.60E-08 3.19E+04 5.00E-04
89C8-23-F2 3.30E-08 2.41E+04 7.86E-04
89C8-45-F2 3.50E-08 1.95E+04 6.77E-04
89C8-98-F2 1.20E-07 7.70E+03 9.37E-04
89C8-23-F3 3.40E-08 2.53E+04 8.67E-04
89C8-45-F3 2.60E-08 2.01E+04 5.29E-04
89C8-98-F3 9.60E-08 8.25E+03 7.88E-04
89C8 7.60E-09 5.04E+04 3.85E-04
ACE2-Fc 2.40E-07 7.34E+04 1.78E-02
另外，AHC传感器在分析缓冲液中线下平衡30min，然后线上检测60s建立基线，在线加载如上所述获得的经纯化的S1抗体/ACE2胞外区(全长)融合蛋白至AHC传感器上。再将传感器放入100nM、50nM、25nM、12.5nM的S1-His抗原中作用3600s，之后将传感器转移至PBS中解离6400s。使用1:1结合模型进行动力学的分析。
实验结果如图2所示，使用全长的ACE2胞外区构建的抗体融合蛋白(优选结构1，F1)和S1的亲和力比89C8高10倍。
实施例3：抗体/ACE2胞外区融合蛋白与细胞表面表达的Spike结合
在本实施例中，检测纯化获得的抗体/ACE2胞外区融合蛋白样品89C8-ACE2-F1与293-Spike细胞结合活性。
通过转染克隆到MCS的SARS-CoV-2Spike蛋白全长基因的PLVX-puro载体产生过表达Spike蛋白的293T细胞(293-Spike细胞)。用胰蛋白酶处理扩大培养的293-Spike细胞，并调整细胞悬液密度至2×106个细胞/ml，100μl/孔加入96孔流式板，使用2％BSA封闭后离心备用将纯化的抗体/ACE2胞外区融合蛋白用PBS稀释，200nM开始3倍稀释共12个点。将上述稀释好的样品100μl/孔加入上述带有细胞的96孔流式板中，室温孵育60min，PBS清洗两次。100μl/孔加入用PBS稀释100倍的羊F(ab’)2抗人IgG-Fc(PE)(购自Abcam)，4℃孵育30min，PBS清洗两次。100μl/孔加入PBS重悬细胞，在CytoFlex(Bechman)流式细胞仪上进行检测并计算对应的MFI。
实验结果如图3所示。可见，89C8-ACE2-F1与293-Spike细胞有很好的结合活性。
实施例4：抗体/ACE2胞外区融合蛋白阻断S1蛋白和ACE2结合的活性
通过转染克隆到MCS的人ACE2蛋白全长基因的PLVX-puro载体产生过表达人ACE2蛋白的CHO细胞(CHO-ACE2细胞)。将扩大培养的CHO-ACE2细胞调整细胞悬液密度至2×106个细胞/ml，100μl/孔加入96孔流式板，使用2％BSA封闭后离心备用。将纯化的样品用PBS稀释，200nM开始3倍稀释共12个点，将上述稀释好的样品60μl/孔加入96孔样品稀释板，同时60μl/孔加入S1-mFc蛋白(购自sinobiologics)，终浓度为0.1nM，与样品37℃孵育。将共孵育样品100μl/孔加入上述带有细胞的96孔流式板中，室温孵育60min，PBS清洗3次。100μl/孔加入用PBS稀释100倍的Anti-mouse Fc-APC(购自eBioscience)，4℃孵育30min，PBS清洗两次。100μl/孔加入PBS重悬细胞，在CytoFlex(Bechman)流式细胞仪上进行检测并计算对应的MFI。
在如上方法的测定实验中，实验结果如图4所示。其结果表明，89C8-ACE2-F1可以阻断ACE2与Spike蛋白的结合，且阻断水平优于对照样品ACE2-Fc及89C8。
实施例5：抗体/ACE2胞外区融合蛋白阻断SARS-Cov2 Spike假病毒侵染ACE2过表达细胞实验
通过转染克隆到MCS的人ACE2蛋白全长基因的PLVX-puro载体产生过表达人ACE2蛋白的293T细胞(293-ACE2细胞)。将293-ACE2细胞按照8000个细胞/孔接种入96孔细胞培养板中，培养过夜。梯度稀释的89C8-ACE2-F1、89C8和ACE2-Fc样品和终浓度为1×105TU/ml的luciferase荧光报告基因的SARS-Cov2 Spike假病毒(购买自北京天坛药物生物技术开发公司)混合，37℃孵育1h。去除293-ACE2细胞培养基，并加入上述样品和假病毒的混合液，37℃孵育48h。检测细胞荧光并计算样品的中和活性。
实验结果如图5所示。
其中，89C8-ACE2-F1、89C8和ACE2-Fc均可以完全中和假病毒的侵染活性，其中89C8和ACE2-Fc的中和活性相似。抗体/ACE2胞外区融合蛋白89C8-ACE2-F1的中和活性比89C8强约60倍。
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。
序列表
<110> 普米斯生物技术（珠海）有限公司
<120> 一种构建冠状病毒抗体的平台
<130> P2020-0837
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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 Gln Ser Ile Lys Val Arg Ile Ser Leu Lys
595 600 605
Ser Ala Leu Gly Asp Lys Ala Tyr Glu Trp Asn Asp Asn Glu Met Tyr
610 615 620
Leu Phe Arg Ser Ser Val Ala Tyr Ala Met Arg Gln Tyr Phe Leu Lys
625 630 635 640
Val Lys Asn Gln Met Ile Leu Phe Gly Glu Glu Asp Val Arg Val Ala
645 650 655
Asn Leu Lys Pro Arg Ile Ser Phe Asn Phe Phe Val Thr Ala Pro Lys
660 665 670
Asn Val Ser Asp Ile Ile Pro Arg Thr Glu Val Glu Lys Ala Ile Arg
675 680 685
Met Ser Arg
690
<210> 5
<211> 127
<212> PRT
<213> 人工序列(artificial sequence)
<400> 5
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr
20 25 30
Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gly Ile Ile Pro Ile Leu Gly Ile Ala Asn Tyr Ala Gln Lys Phe
50 55 60
Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Gly Leu Gly Phe Gly Glu Leu Phe Arg Lys Thr Gly Tyr
100 105 110
Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 125
<210> 6
<211> 112
<212> PRT
<213> 人工序列(artificial sequence)
<400> 6
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Ala Ala Ser Thr Arg Ala Thr Ala Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Val Tyr Phe Cys Gln Gln Tyr Asn Lys Leu Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Asp Ile Lys Arg Thr Val Ala Ala
100 105 110
<210> 7
<211> 226
<212> PRT
<213> 人工序列(artificial sequence)
<400> 7
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly
225
<210> 8
<211> 107
<212> PRT
<213> 人工序列(artificial sequence)
<400> 8
Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
1 5 10 15
Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
20 25 30
Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
35 40 45
Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
50 55 60
Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu
65 70 75 80
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
85 90 95
Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
100 105
<210> 9
<211> 103
<212> PRT
<213> 人工序列(artificial sequence)
<400> 9
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys
1 5 10 15
Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
20 25 30
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
35 40 45
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
50 55 60
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
65 70 75 80
Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95
Lys Val Glu Pro Lys Ser Cys
100
<210> 10
<211> 21
<212> PRT
<213> 人工序列(artificial sequence)
<400> 10
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
1 5 10 15
Gly Gly Gly Ser Gly
20
<210> 11
<211> 456
<212> PRT
<213> 人工序列(artificial sequence)
<400> 11
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr
20 25 30
Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gly Ile Ile Pro Ile Leu Gly Ile Ala Asn Tyr Ala Gln Lys Phe
50 55 60
Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Gly Leu Gly Phe Gly Glu Leu Phe Arg Lys Thr Gly Tyr
100 105 110
Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala
115 120 125
Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser
130 135 140
Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe
145 150 155 160
Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly
165 170 175
Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu
180 185 190
Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr
195 200 205
Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys
210 215 220
Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro
225 230 235 240
Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys
245 250 255
Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val
260 265 270
Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr
275 280 285
Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu
290 295 300
Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His
305 310 315 320
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys
325 330 335
Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln
340 345 350
Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met
355 360 365
Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro
370 375 380
Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn
385 390 395 400
Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu
405 410 415
Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val
420 425 430
Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln
435 440 445
Lys Ser Leu Ser Leu Ser Pro Gly
450 455
<210> 12
<211> 498
<212> PRT
<213> 人工序列(artificial sequence)
<400> 12
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr
20 25 30
Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gly Ile Ile Pro Ile Leu Gly Ile Ala Asn Tyr Ala Gln Lys Phe
50 55 60
Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Gly Leu Gly Phe Gly Glu Leu Phe Arg Lys Thr Gly Tyr
100 105 110
Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala
115 120 125
Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser
130 135 140
Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe
145 150 155 160
Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly
165 170 175
Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu
180 185 190
Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr
195 200 205
Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys
210 215 220
Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro
225 230 235 240
Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys
245 250 255
Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val
260 265 270
Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr
275 280 285
Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu
290 295 300
Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His
305 310 315 320
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys
325 330 335
Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln
340 345 350
Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met
355 360 365
Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro
370 375 380
Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn
385 390 395 400
Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu
405 410 415
Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val
420 425 430
Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln
435 440 445
Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly
450 455 460
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Glu Gln Ala
465 470 475 480
Lys Thr Phe Leu Asp Lys Phe Asn His Glu Ala Glu Asp Leu Phe Tyr
485 490 495
Gln Ser
<210> 13
<211> 522
<212> PRT
<213> 人工序列(artificial sequence)
<400> 13
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr
20 25 30
Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gly Ile Ile Pro Ile Leu Gly Ile Ala Asn Tyr Ala Gln Lys Phe
50 55 60
Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Gly Leu Gly Phe Gly Glu Leu Phe Arg Lys Thr Gly Tyr
100 105 110
Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala
115 120 125
Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser
130 135 140
Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe
145 150 155 160
Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly
165 170 175
Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu
180 185 190
Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr
195 200 205
Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys
210 215 220
Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro
225 230 235 240
Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys
245 250 255
Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val
260 265 270
Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr
275 280 285
Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu
290 295 300
Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His
305 310 315 320
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys
325 330 335
Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln
340 345 350
Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met
355 360 365
Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro
370 375 380
Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn
385 390 395 400
Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu
405 410 415
Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val
420 425 430
Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln
435 440 445
Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly
450 455 460
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gln Ser Thr
465 470 475 480
Ile Glu Glu Gln Ala Lys Thr Phe Leu Asp Lys Phe Asn His Glu Ala
485 490 495
Glu Asp Leu Phe Tyr Gln Ser Ser Leu Ala Ser Trp Asn Tyr Asn Thr
500 505 510
Asn Ile Thr Glu Glu Asn Val Gln Asn Met
515 520
<210> 14
<211> 575
<212> PRT
<213> 人工序列(artificial sequence)
<400> 14
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr
20 25 30
Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gly Ile Ile Pro Ile Leu Gly Ile Ala Asn Tyr Ala Gln Lys Phe
50 55 60
Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Gly Leu Gly Phe Gly Glu Leu Phe Arg Lys Thr Gly Tyr
100 105 110
Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala
115 120 125
Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser
130 135 140
Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe
145 150 155 160
Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly
165 170 175
Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu
180 185 190
Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr
195 200 205
Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys
210 215 220
Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro
225 230 235 240
Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys
245 250 255
Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val
260 265 270
Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr
275 280 285
Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu
290 295 300
Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His
305 310 315 320
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys
325 330 335
Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln
340 345 350
Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met
355 360 365
Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro
370 375 380
Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn
385 390 395 400
Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu
405 410 415
Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val
420 425 430
Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln
435 440 445
Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly
450 455 460
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gln Ser Thr
465 470 475 480
Ile Glu Glu Gln Ala Lys Thr Phe Leu Asp Lys Phe Asn His Glu Ala
485 490 495
Glu Asp Leu Phe Tyr Gln Ser Ser Leu Ala Ser Trp Asn Tyr Asn Thr
500 505 510
Asn Ile Thr Glu Glu Asn Val Gln Asn Met Asn Asn Ala Gly Asp Lys
515 520 525
Trp Ser Ala Phe Leu Lys Glu Gln Ser Thr Leu Ala Gln Met Tyr Pro
530 535 540
Leu Gln Glu Ile Gln Asn Leu Thr Val Lys Leu Gln Leu Gln Ala Leu
545 550 555 560
Gln Gln Asn Gly Ser Ser Val Leu Ser Glu Asp Lys Ser Lys Arg
565 570 575
<210> 15
<211> 1168
<212> PRT
<213> 人工序列(artificial sequence)
<400> 15
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr
20 25 30
Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gly Ile Ile Pro Ile Leu Gly Ile Ala Asn Tyr Ala Gln Lys Phe
50 55 60
Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Gly Leu Gly Phe Gly Glu Leu Phe Arg Lys Thr Gly Tyr
100 105 110
Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala
115 120 125
Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser
130 135 140
Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe
145 150 155 160
Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly
165 170 175
Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu
180 185 190
Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr
195 200 205
Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys
210 215 220
Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro
225 230 235 240
Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys
245 250 255
Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val
260 265 270
Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr
275 280 285
Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu
290 295 300
Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His
305 310 315 320
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys
325 330 335
Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln
340 345 350
Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met
355 360 365
Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro
370 375 380
Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn
385 390 395 400
Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu
405 410 415
Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val
420 425 430
Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln
435 440 445
Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly
450 455 460
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gln Ser Thr
465 470 475 480
Ile Glu Glu Gln Ala Lys Thr Phe Leu Asp Lys Phe Asn His Glu Ala
485 490 495
Glu Asp Leu Phe Tyr Gln Ser Ser Leu Ala Ser Trp Asn Tyr Asn Thr
500 505 510
Asn Ile Thr Glu Glu Asn Val Gln Asn Met Asn Asn Ala Gly Asp Lys
515 520 525
Trp Ser Ala Phe Leu Lys Glu Gln Ser Thr Leu Ala Gln Met Tyr Pro
530 535 540
Leu Gln Glu Ile Gln Asn Leu Thr Val Lys Leu Gln Leu Gln Ala Leu
545 550 555 560
Gln Gln Asn Gly Ser Ser Val Leu Ser Glu Asp Lys Ser Lys Arg Leu
565 570 575
Asn Thr Ile Leu Asn Thr Met Ser Thr Ile Tyr Ser Thr Gly Lys Val
580 585 590
Cys Asn Pro Asp Asn Pro Gln Glu Cys Leu Leu Leu Glu Pro Gly Leu
595 600 605
Asn Glu Ile Met Ala Asn Ser Leu Asp Tyr Asn Glu Arg Leu Trp Ala
610 615 620
Trp Glu Ser Trp Arg Ser Glu Val Gly Lys Gln Leu Arg Pro Leu Tyr
625 630 635 640
Glu Glu Tyr Val Val Leu Lys Asn Glu Met Ala Arg Ala Asn His Tyr
645 650 655
Glu Asp Tyr Gly Asp Tyr Trp Arg Gly Asp Tyr Glu Val Asn Gly Val
660 665 670
Asp Gly Tyr Asp Tyr Ser Arg Gly Gln Leu Ile Glu Asp Val Glu His
675 680 685
Thr Phe Glu Glu Ile Lys Pro Leu Tyr Glu His Leu His Ala Tyr Val
690 695 700
Arg Ala Lys Leu Met Asn Ala Tyr Pro Ser Tyr Ile Ser Pro Ile Gly
705 710 715 720
Cys Leu Pro Ala His Leu Leu Gly Asp Met Trp Gly Arg Phe Trp Thr
725 730 735
Asn Leu Tyr Ser Leu Thr Val Pro Phe Gly Gln Lys Pro Asn Ile Asp
740 745 750
Val Thr Asp Ala Met Val Asp Gln Ala Trp Asp Ala Gln Arg Ile Phe
755 760 765
Lys Glu Ala Glu Lys Phe Phe Val Ser Val Gly Leu Pro Asn Met Thr
770 775 780
Gln Gly Phe Trp Glu Asn Ser Met Leu Thr Asp Pro Gly Asn Val Gln
785 790 795 800
Lys Ala Val Cys His Pro Thr Ala Trp Asp Leu Gly Lys Gly Asp Phe
805 810 815
Arg Ile Leu Met Cys Thr Lys Val Thr Met Asp Asp Phe Leu Thr Ala
820 825 830
His His Glu Met Gly His Ile Gln Tyr Asp Met Ala Tyr Ala Ala Gln
835 840 845
Pro Phe Leu Leu Arg Asn Gly Ala Asn Glu Gly Phe His Glu Ala Val
850 855 860
Gly Glu Ile Met Ser Leu Ser Ala Ala Thr Pro Lys His Leu Lys Ser
865 870 875 880
Ile Gly Leu Leu Ser Pro Asp Phe Gln Glu Asp Asn Glu Thr Glu Ile
885 890 895
Asn Phe Leu Leu Lys Gln Ala Leu Thr Ile Val Gly Thr Leu Pro Phe
900 905 910
Thr Tyr Met Leu Glu Lys Trp Arg Trp Met Val Phe Lys Gly Glu Ile
915 920 925
Pro Lys Asp Gln Trp Met Lys Lys Trp Trp Glu Met Lys Arg Glu Ile
930 935 940
Val Gly Val Val Glu Pro Val Pro His Asp Glu Thr Tyr Cys Asp Pro
945 950 955 960
Ala Ser Leu Phe His Val Ser Asn Asp Tyr Ser Phe Ile Arg Tyr Tyr
965 970 975
Thr Arg Thr Leu Tyr Gln Phe Gln Phe Gln Glu Ala Leu Cys Gln Ala
980 985 990
Ala Lys His Glu Gly Pro Leu His Lys Cys Asp Ile Ser Asn Ser Thr
995 1000 1005
Glu Ala Gly Gln Lys Leu Phe Asn Met Leu Arg Leu Gly Lys Ser
1010 1015 1020
Glu Pro Trp Thr Leu Ala Leu Glu Asn Val Val Gly Ala Lys Asn
1025 1030 1035
Met Asn Val Arg Pro Leu Leu Asn Tyr Phe Glu Pro Leu Phe Thr
1040 1045 1050
Trp Leu Lys Asp Gln Asn Lys Asn Ser Phe Val Gly Trp Ser Thr
1055 1060 1065
Asp Trp Ser Pro Tyr Ala Asp Gln Ser Ile Lys Val Arg Ile Ser
1070 1075 1080
Leu Lys Ser Ala Leu Gly Asp Lys Ala Tyr Glu Trp Asn Asp Asn
1085 1090 1095
Glu Met Tyr Leu Phe Arg Ser Ser Val Ala Tyr Ala Met Arg Gln
1100 1105 1110
Tyr Phe Leu Lys Val Lys Asn Gln Met Ile Leu Phe Gly Glu Glu
1115 1120 1125
Asp Val Arg Val Ala Asn Leu Lys Pro Arg Ile Ser Phe Asn Phe
1130 1135 1140
Phe Val Thr Ala Pro Lys Asn Val Ser Asp Ile Ile Pro Arg Thr
1145 1150 1155
Glu Val Glu Lys Ala Ile Arg Met Ser Arg
1160 1165
<210> 16
<211> 219
<212> PRT
<213> 人工序列(artificial sequence)
<400> 16
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Ala Ala Ser Thr Arg Ala Thr Ala Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Val Tyr Phe Cys Gln Gln Tyr Asn Lys Leu Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Asp Ile Lys Arg Thr Val Ala Ala
100 105 110
Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
115 120 125
Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
130 135 140
Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
145 150 155 160
Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
165 170 175
Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu
180 185 190
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
195 200 205
Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 17
<211> 261
<212> PRT
<213> 人工序列(artificial sequence)
<400> 17
Glu Gln Ala Lys Thr Phe Leu Asp Lys Phe Asn His Glu Ala Glu Asp
1 5 10 15
Leu Phe Tyr Gln Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
20 25 30
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Glu Ile Val Met Thr Gln
35 40 45
Ser Pro Ala Thr Leu Ser Val Ser Pro Gly Glu Arg Ala Thr Leu Ser
50 55 60
Cys Arg Ala Ser Gln Ser Val Ser Ser Asn Leu Ala Trp Tyr Gln Gln
65 70 75 80
Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr Ala Ala Ser Thr Arg
85 90 95
Ala Thr Ala Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu
100 105 110
Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr
115 120 125
Phe Cys Gln Gln Tyr Asn Lys Leu Pro Leu Thr Phe Gly Gly Gly Thr
130 135 140
Lys Val Asp Ile Lys Arg Thr Val Ala Ala Arg Thr Val Ala Ala Pro
145 150 155 160
Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr
165 170 175
Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys
180 185 190
Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu
195 200 205
Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser
210 215 220
Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala
225 230 235 240
Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe
245 250 255
Asn Arg Gly Glu Cys
260
<210> 18
<211> 285
<212> PRT
<213> 人工序列(artificial sequence)
<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 Gly Gly Gly
35 40 45
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
50 55 60
Ser Gly Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser
65 70 75 80
Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser
85 90 95
Ser Asn Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu
100 105 110
Leu Ile Tyr Ala Ala Ser Thr Arg Ala Thr Ala Ile Pro Ala Arg Phe
115 120 125
Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu
130 135 140
Gln Ser Glu Asp Phe Ala Val Tyr Phe Cys Gln Gln Tyr Asn Lys Leu
145 150 155 160
Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Asp Ile Lys Arg Thr Val
165 170 175
Ala Ala Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser
180 185 190
Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn
195 200 205
Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala
210 215 220
Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys
225 230 235 240
Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp
245 250 255
Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu
260 265 270
Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
275 280 285
<210> 19
<211> 338
<212> PRT
<213> 人工序列(artificial sequence)
<400> 19
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 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
100 105 110
Ser Gly Gly Gly Gly Ser Gly Glu Ile Val Met Thr Gln Ser Pro Ala
115 120 125
Thr Leu Ser Val Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala
130 135 140
Ser Gln Ser Val Ser Ser Asn Leu Ala Trp Tyr Gln Gln Lys Pro Gly
145 150 155 160
Gln Ala Pro Arg Leu Leu Ile Tyr Ala Ala Ser Thr Arg Ala Thr Ala
165 170 175
Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu
180 185 190
Thr Ile Ser Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr Phe Cys Gln
195 200 205
Gln Tyr Asn Lys Leu Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Asp
210 215 220
Ile Lys Arg Thr Val Ala Ala Arg Thr Val Ala Ala Pro Ser Val Phe
225 230 235 240
Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val
245 250 255
Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp
260 265 270
Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr
275 280 285
Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr
290 295 300
Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val
305 310 315 320
Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly
325 330 335
Glu Cys
<210> 20
<211> 931
<212> PRT
<213> 人工序列(artificial sequence)
<400> 20
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 Gln Ser Ile Lys Val Arg Ile Ser Leu Lys
595 600 605
Ser Ala Leu Gly Asp Lys Ala Tyr Glu Trp Asn Asp Asn Glu Met Tyr
610 615 620
Leu Phe Arg Ser Ser Val Ala Tyr Ala Met Arg Gln Tyr Phe Leu Lys
625 630 635 640
Val Lys Asn Gln Met Ile Leu Phe Gly Glu Glu Asp Val Arg Val Ala
645 650 655
Asn Leu Lys Pro Arg Ile Ser Phe Asn Phe Phe Val Thr Ala Pro Lys
660 665 670
Asn Val Ser Asp Ile Ile Pro Arg Thr Glu Val Glu Lys Ala Ile Arg
675 680 685
Met Ser Arg Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
690 695 700
Gly Ser Gly Gly Gly Gly Ser Gly Glu Ile Val Met Thr Gln Ser Pro
705 710 715 720
Ala Thr Leu Ser Val Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg
725 730 735
Ala Ser Gln Ser Val Ser Ser Asn Leu Ala Trp Tyr Gln Gln Lys Pro
740 745 750
Gly Gln Ala Pro Arg Leu Leu Ile Tyr Ala Ala Ser Thr Arg Ala Thr
755 760 765
Ala Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr
770 775 780
Leu Thr Ile Ser Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr Phe Cys
785 790 795 800
Gln Gln Tyr Asn Lys Leu Pro Leu Thr Phe Gly Gly Gly Thr Lys Val
805 810 815
Asp Ile Lys Arg Thr Val Ala Ala Arg Thr Val Ala Ala Pro Ser Val
820 825 830
Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser
835 840 845
Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln
850 855 860
Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val
865 870 875 880
Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu
885 890 895
Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu
900 905 910
Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg
915 920 925
Gly Glu Cys
930
<210> 21
<211> 448
<212> PRT
<213> 人工序列(artificial sequence)
<400> 21
Glu Val Gln Leu Val Gln Ser Gly Thr Glu Val Lys Lys Pro Gly Glu
1 5 10 15
Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Gly Phe Ile Thr Tyr
20 25 30
Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met
35 40 45
Gly Ile Ile Tyr Pro Gly Asp Ser Glu Thr Arg Tyr Ser Pro Ser Phe
50 55 60
Gln Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Asn Thr Ala Tyr
65 70 75 80
Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Ile Tyr Tyr Cys
85 90 95
Ala Gly Gly Ser Gly Ile Ser Thr Pro Met Asp Val Trp Gly Gln Gly
100 105 110
Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
225 230 235 240
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
340 345 350
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
355 360 365
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
<210> 22
<211> 220
<212> PRT
<213> 人工序列(artificial sequence)
<400> 22
Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Tyr Ser
20 25 30
Ser Ile Asn Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln
35 40 45
Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
50 55 60
Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
65 70 75 80
Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln
85 90 95
Tyr Tyr Ser Thr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile
100 105 110
Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp
115 120 125
Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn
130 135 140
Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu
145 150 155 160
Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp
165 170 175
Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr
180 185 190
Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser
195 200 205
Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
210 215 220
<210> 23
<211> 490
<212> PRT
<213> 人工序列(artificial sequence)
<400> 23
Glu Val Gln Leu Val Gln Ser Gly Thr Glu Val Lys Lys Pro Gly Glu
1 5 10 15
Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Gly Phe Ile Thr Tyr
20 25 30
Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met
35 40 45
Gly Ile Ile Tyr Pro Gly Asp Ser Glu Thr Arg Tyr Ser Pro Ser Phe
50 55 60
Gln Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Asn Thr Ala Tyr
65 70 75 80
Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Ile Tyr Tyr Cys
85 90 95
Ala Gly Gly Ser Gly Ile Ser Thr Pro Met Asp Val Trp Gly Gln Gly
100 105 110
Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro
225 230 235 240
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
340 345 350
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
355 360 365
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
450 455 460
Gly Gly Gly Ser Gly Glu Gln Ala Lys Thr Phe Leu Asp Lys Phe Asn
465 470 475 480
His Glu Ala Glu Asp Leu Phe Tyr Gln Ser
485 490
<210> 24
<211> 514
<212> PRT
<213> 人工序列(artificial sequence)
<400> 24
Glu Val Gln Leu Val Gln Ser Gly Thr Glu Val Lys Lys Pro Gly Glu
1 5 10 15
Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Gly Phe Ile Thr Tyr
20 25 30
Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met
35 40 45
Gly Ile Ile Tyr Pro Gly Asp Ser Glu Thr Arg Tyr Ser Pro Ser Phe
50 55 60
Gln Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Asn Thr Ala Tyr
65 70 75 80
Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Ile Tyr Tyr Cys
85 90 95
Ala Gly Gly Ser Gly Ile Ser Thr Pro Met Asp Val Trp Gly Gln Gly
100 105 110
Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro
225 230 235 240
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
340 345 350
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
355 360 365
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
450 455 460
Gly Gly Gly Ser Gly Gln Ser Thr Ile Glu Glu Gln Ala Lys Thr Phe
465 470 475 480
Leu Asp Lys Phe Asn His Glu Ala Glu Asp Leu Phe Tyr Gln Ser Ser
485 490 495
Leu Ala Ser Trp Asn Tyr Asn Thr Asn Ile Thr Glu Glu Asn Val Gln
500 505 510
Asn Met
<210> 25
<211> 567
<212> PRT
<213> 人工序列(artificial sequence)
<400> 25
Glu Val Gln Leu Val Gln Ser Gly Thr Glu Val Lys Lys Pro Gly Glu
1 5 10 15
Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Gly Phe Ile Thr Tyr
20 25 30
Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met
35 40 45
Gly Ile Ile Tyr Pro Gly Asp Ser Glu Thr Arg Tyr Ser Pro Ser Phe
50 55 60
Gln Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Asn Thr Ala Tyr
65 70 75 80
Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Ile Tyr Tyr Cys
85 90 95
Ala Gly Gly Ser Gly Ile Ser Thr Pro Met Asp Val Trp Gly Gln Gly
100 105 110
Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro
225 230 235 240
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
340 345 350
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
355 360 365
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
450 455 460
Gly Gly Gly Ser Gly Gln Ser Thr Ile Glu Glu Gln Ala Lys Thr Phe
465 470 475 480
Leu Asp Lys Phe Asn His Glu Ala Glu Asp Leu Phe Tyr Gln Ser Ser
485 490 495
Leu Ala Ser Trp Asn Tyr Asn Thr Asn Ile Thr Glu Glu Asn Val Gln
500 505 510
Asn Met Asn Asn Ala Gly Asp Lys Trp Ser Ala Phe Leu Lys Glu Gln
515 520 525
Ser Thr Leu Ala Gln Met Tyr Pro Leu Gln Glu Ile Gln Asn Leu Thr
530 535 540
Val Lys Leu Gln Leu Gln Ala Leu Gln Gln Asn Gly Ser Ser Val Leu
545 550 555 560
Ser Glu Asp Lys Ser Lys Arg
565
<210> 26
<211> 1160
<212> PRT
<213> 人工序列(artificial sequence)
<400> 26
Glu Val Gln Leu Val Gln Ser Gly Thr Glu Val Lys Lys Pro Gly Glu
1 5 10 15
Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Gly Phe Ile Thr Tyr
20 25 30
Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met
35 40 45
Gly Ile Ile Tyr Pro Gly Asp Ser Glu Thr Arg Tyr Ser Pro Ser Phe
50 55 60
Gln Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Asn Thr Ala Tyr
65 70 75 80
Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Ile Tyr Tyr Cys
85 90 95
Ala Gly Gly Ser Gly Ile Ser Thr Pro Met Asp Val Trp Gly Gln Gly
100 105 110
Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro
225 230 235 240
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
340 345 350
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
355 360 365
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
450 455 460
Gly Gly Gly Ser Gly Gln Ser Thr Ile Glu Glu Gln Ala Lys Thr Phe
465 470 475 480
Leu Asp Lys Phe Asn His Glu Ala Glu Asp Leu Phe Tyr Gln Ser Ser
485 490 495
Leu Ala Ser Trp Asn Tyr Asn Thr Asn Ile Thr Glu Glu Asn Val Gln
500 505 510
Asn Met Asn Asn Ala Gly Asp Lys Trp Ser Ala Phe Leu Lys Glu Gln
515 520 525
Ser Thr Leu Ala Gln Met Tyr Pro Leu Gln Glu Ile Gln Asn Leu Thr
530 535 540
Val Lys Leu Gln Leu Gln Ala Leu Gln Gln Asn Gly Ser Ser Val Leu
545 550 555 560
Ser Glu Asp Lys Ser Lys Arg Leu Asn Thr Ile Leu Asn Thr Met Ser
565 570 575
Thr Ile Tyr Ser Thr Gly Lys Val Cys Asn Pro Asp Asn Pro Gln Glu
580 585 590
Cys Leu Leu Leu Glu Pro Gly Leu Asn Glu Ile Met Ala Asn Ser Leu
595 600 605
Asp Tyr Asn Glu Arg Leu Trp Ala Trp Glu Ser Trp Arg Ser Glu Val
610 615 620
Gly Lys Gln Leu Arg Pro Leu Tyr Glu Glu Tyr Val Val Leu Lys Asn
625 630 635 640
Glu Met Ala Arg Ala Asn His Tyr Glu Asp Tyr Gly Asp Tyr Trp Arg
645 650 655
Gly Asp Tyr Glu Val Asn Gly Val Asp Gly Tyr Asp Tyr Ser Arg Gly
660 665 670
Gln Leu Ile Glu Asp Val Glu His Thr Phe Glu Glu Ile Lys Pro Leu
675 680 685
Tyr Glu His Leu His Ala Tyr Val Arg Ala Lys Leu Met Asn Ala Tyr
690 695 700
Pro Ser Tyr Ile Ser Pro Ile Gly Cys Leu Pro Ala His Leu Leu Gly
705 710 715 720
Asp Met Trp Gly Arg Phe Trp Thr Asn Leu Tyr Ser Leu Thr Val Pro
725 730 735
Phe Gly Gln Lys Pro Asn Ile Asp Val Thr Asp Ala Met Val Asp Gln
740 745 750
Ala Trp Asp Ala Gln Arg Ile Phe Lys Glu Ala Glu Lys Phe Phe Val
755 760 765
Ser Val Gly Leu Pro Asn Met Thr Gln Gly Phe Trp Glu Asn Ser Met
770 775 780
Leu Thr Asp Pro Gly Asn Val Gln Lys Ala Val Cys His Pro Thr Ala
785 790 795 800
Trp Asp Leu Gly Lys Gly Asp Phe Arg Ile Leu Met Cys Thr Lys Val
805 810 815
Thr Met Asp Asp Phe Leu Thr Ala His His Glu Met Gly His Ile Gln
820 825 830
Tyr Asp Met Ala Tyr Ala Ala Gln Pro Phe Leu Leu Arg Asn Gly Ala
835 840 845
Asn Glu Gly Phe His Glu Ala Val Gly Glu Ile Met Ser Leu Ser Ala
850 855 860
Ala Thr Pro Lys His Leu Lys Ser Ile Gly Leu Leu Ser Pro Asp Phe
865 870 875 880
Gln Glu Asp Asn Glu Thr Glu Ile Asn Phe Leu Leu Lys Gln Ala Leu
885 890 895
Thr Ile Val Gly Thr Leu Pro Phe Thr Tyr Met Leu Glu Lys Trp Arg
900 905 910
Trp Met Val Phe Lys Gly Glu Ile Pro Lys Asp Gln Trp Met Lys Lys
915 920 925
Trp Trp Glu Met Lys Arg Glu Ile Val Gly Val Val Glu Pro Val Pro
930 935 940
His Asp Glu Thr Tyr Cys Asp Pro Ala Ser Leu Phe His Val Ser Asn
945 950 955 960
Asp Tyr Ser Phe Ile Arg Tyr Tyr Thr Arg Thr Leu Tyr Gln Phe Gln
965 970 975
Phe Gln Glu Ala Leu Cys Gln Ala Ala Lys His Glu Gly Pro Leu His
980 985 990
Lys Cys Asp Ile Ser Asn Ser Thr Glu Ala Gly Gln Lys Leu Phe Asn
995 1000 1005
Met Leu Arg Leu Gly Lys Ser Glu Pro Trp Thr Leu Ala Leu Glu
1010 1015 1020
Asn Val Val Gly Ala Lys Asn Met Asn Val Arg Pro Leu Leu Asn
1025 1030 1035
Tyr Phe Glu Pro Leu Phe Thr Trp Leu Lys Asp Gln Asn Lys Asn
1040 1045 1050
Ser Phe Val Gly Trp Ser Thr Asp Trp Ser Pro Tyr Ala Asp Gln
1055 1060 1065
Ser Ile Lys Val Arg Ile Ser Leu Lys Ser Ala Leu Gly Asp Lys
1070 1075 1080
Ala Tyr Glu Trp Asn Asp Asn Glu Met Tyr Leu Phe Arg Ser Ser
1085 1090 1095
Val Ala Tyr Ala Met Arg Gln Tyr Phe Leu Lys Val Lys Asn Gln
1100 1105 1110
Met Ile Leu Phe Gly Glu Glu Asp Val Arg Val Ala Asn Leu Lys
1115 1120 1125
Pro Arg Ile Ser Phe Asn Phe Phe Val Thr Ala Pro Lys Asn Val
1130 1135 1140
Ser Asp Ile Ile Pro Arg Thr Glu Val Glu Lys Ala Ile Arg Met
1145 1150 1155
Ser Arg
1160
<210> 27
<211> 267
<212> PRT
<213> 人工序列(artificial sequence)
<400> 27
Glu Gln Ala Lys Thr Phe Leu Asp Lys Phe Asn His Glu Ala Glu Asp
1 5 10 15
Leu Phe Tyr Gln Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
20 25 30
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Asp Ile Val Met Thr Gln
35 40 45
Ser Pro Asp Ser Leu Ala Val Ser Leu Gly Glu Arg Ala Thr Ile Asn
50 55 60
Cys Lys Ser Ser Gln Ser Val Leu Tyr Ser Ser Ile Asn Lys Asn Tyr
65 70 75 80
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile
85 90 95
Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Ser Gly
100 105 110
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala
115 120 125
Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Tyr Tyr Ser Thr Pro Tyr
130 135 140
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala
145 150 155 160
Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
165 170 175
Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
180 185 190
Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
195 200 205
Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
210 215 220
Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu
225 230 235 240
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
245 250 255
Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
260 265
<210> 28
<211> 291
<212> PRT
<213> 人工序列(artificial sequence)
<400> 28
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 Gly Gly Gly
35 40 45
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
50 55 60
Ser Gly Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser
65 70 75 80
Leu Gly Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu
85 90 95
Tyr Ser Ser Ile Asn Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro
100 105 110
Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser
115 120 125
Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
130 135 140
Leu Thr Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys
145 150 155 160
Gln Gln Tyr Tyr Ser Thr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val
165 170 175
Glu Ile Lys Arg Thr Val Ala Ala Arg Thr Val Ala Ala Pro Ser Val
180 185 190
Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser
195 200 205
Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln
210 215 220
Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val
225 230 235 240
Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu
245 250 255
Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu
260 265 270
Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg
275 280 285
Gly Glu Cys
290
<210> 29
<211> 344
<212> PRT
<213> 人工序列(artificial sequence)
<400> 29
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 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
100 105 110
Ser Gly Gly Gly Gly Ser Gly Asp Ile Val Met Thr Gln Ser Pro Asp
115 120 125
Ser Leu Ala Val Ser Leu Gly Glu Arg Ala Thr Ile Asn Cys Lys Ser
130 135 140
Ser Gln Ser Val Leu Tyr Ser Ser Ile Asn Lys Asn Tyr Leu Ala Trp
145 150 155 160
Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala
165 170 175
Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser
180 185 190
Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala Glu Asp Val
195 200 205
Ala Val Tyr Tyr Cys Gln Gln Tyr Tyr Ser Thr Pro Tyr Thr Phe Gly
210 215 220
Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Arg Thr Val
225 230 235 240
Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys
245 250 255
Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg
260 265 270
Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn
275 280 285
Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser
290 295 300
Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys
305 310 315 320
Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr
325 330 335
Lys Ser Phe Asn Arg Gly Glu Cys
340
<210> 30
<211> 937
<212> PRT
<213> 人工序列(artificial sequence)
<400> 30
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 Gln Ser Ile Lys Val Arg Ile Ser Leu Lys
595 600 605
Ser Ala Leu Gly Asp Lys Ala Tyr Glu Trp Asn Asp Asn Glu Met Tyr
610 615 620
Leu Phe Arg Ser Ser Val Ala Tyr Ala Met Arg Gln Tyr Phe Leu Lys
625 630 635 640
Val Lys Asn Gln Met Ile Leu Phe Gly Glu Glu Asp Val Arg Val Ala
645 650 655
Asn Leu Lys Pro Arg Ile Ser Phe Asn Phe Phe Val Thr Ala Pro Lys
660 665 670
Asn Val Ser Asp Ile Ile Pro Arg Thr Glu Val Glu Lys Ala Ile Arg
675 680 685
Met Ser Arg Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
690 695 700
Gly Ser Gly Gly Gly Gly Ser Gly Asp Ile Val Met Thr Gln Ser Pro
705 710 715 720
Asp Ser Leu Ala Val Ser Leu Gly Glu Arg Ala Thr Ile Asn Cys Lys
725 730 735
Ser Ser Gln Ser Val Leu Tyr Ser Ser Ile Asn Lys Asn Tyr Leu Ala
740 745 750
Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp
755 760 765
Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly
770 775 780
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala Glu Asp
785 790 795 800
Val Ala Val Tyr Tyr Cys Gln Gln Tyr Tyr Ser Thr Pro Tyr Thr Phe
805 810 815
Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Arg Thr
820 825 830
Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu
835 840 845
Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro
850 855 860
Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly
865 870 875 880
Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr
885 890 895
Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His
900 905 910
Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val
915 920 925
Thr Lys Ser Phe Asn Arg Gly Glu Cys
930 935
<210> 31
<211> 917
<212> PRT
<213> 人工序列(artificial sequence)
<400> 31
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 Gln Ser Ile Lys Val Arg Ile Ser Leu Lys
595 600 605
Ser Ala Leu Gly Asp Lys Ala Tyr Glu Trp Asn Asp Asn Glu Met Tyr
610 615 620
Leu Phe Arg Ser Ser Val Ala Tyr Ala Met Arg Gln Tyr Phe Leu Lys
625 630 635 640
Val Lys Asn Gln Met Ile Leu Phe Gly Glu Glu Asp Val Arg Val Ala
645 650 655
Asn Leu Lys Pro Arg Ile Ser Phe Asn Phe Phe Val Thr Ala Pro Lys
660 665 670
Asn Val Ser Asp Ile Ile Pro Arg Thr Glu Val Glu Lys Ala Ile Arg
675 680 685
Met Ser Arg Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu
690 695 700
Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp
705 710 715 720
Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
725 730 735
Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly
740 745 750
Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn
755 760 765
Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp
770 775 780
Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro
785 790 795 800
Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu
805 810 815
Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn
820 825 830
Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile
835 840 845
Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr
850 855 860
Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys
865 870 875 880
Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys
885 890 895
Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu
900 905 910
Ser Leu Ser Pro Gly
915

Claims (11)

1.一种抗冠状病毒的融合蛋白的制备方法，其特征在于，包括步骤：将一抗冠状病毒抗体与人血管紧张素转化酶2(ACE2)或其片段进行融合，从而得到所述融合蛋白。
2.如权利要求1所述的方法，其特征在于，所述的人ACE2或其片段是人ACE2的胞外区全长蛋白或其片段蛋白。
3.如权利要求1所述的方法，其特征在于，所述的融合蛋白中，所述的人ACE2或其片段被融合至所述抗体的重链的N端、重链的C端、轻链的N端，和/或轻链的C端。
4.如权利要求1所述的方法，其特征在于，所述融合蛋白中的H链具有如SEQ ID NO:15所示的氨基酸序列，并且融合蛋白中的L链具有如SEQ ID NO:16所示的氨基酸序列；
或者，所述融合蛋白中的H链具有如SEQ ID NO:15所示的氨基酸序列，并且融合蛋白中的L链具有如SEQ ID NO:20所示的氨基酸序列；
或者，所述融合蛋白中的H链具有如SEQ ID NO:11所示的氨基酸序列，并且融合蛋白中的L链具有如SEQ ID NO:20所示的氨基酸序列。
5.如权利要求1所述的方法，其特征在于，所述融合蛋白中的H链具有如SEQ ID NO:26所示的氨基酸序列，并且融合蛋白中的L链具有如SEQ ID NO:22所示的氨基酸序列；
或者，所述融合蛋白中的H链具有如SEQ ID NO:26所示的氨基酸序列，并且融合蛋白中的L链具有如SEQ ID NO:30所示的氨基酸序列；
或者，所述融合蛋白中的H链具有如SEQ ID NO:21所示的氨基酸序列，并且融合蛋白中的L链具有如SEQ ID NO:30所示的氨基酸序列。
6.一种融合蛋白，其特征在于，所述的融合蛋白的序列中包含：
(a)抗SARS-Cov2 Spike 1蛋白抗体的序列；和
(b)人ACE2或其片段的序列。
7.一种抗体偶联物，其特征在于，所述抗体偶联物含有：
(a)靶向部分，所述靶向部分包括如权利要求6所述的融合蛋白；和
(b)与所述靶向部分偶联的偶联部分，所述偶联部分选自下组：可检测标记物、药物、毒素、细胞因子、放射性核素、酶、金纳米颗粒/纳米棒、纳米磁粒、病毒外壳蛋白或VLP，或其组合。
8.一种免疫细胞，其特征在于，所述免疫细胞表达或在细胞膜外暴露有如权利要求6所述的融合蛋白。
9.一种药物组合物，其特征在于，所述药物组合物含有：
(i)活性成分，所述活性成分选自下组：如权利要求6所述的融合蛋白、如权利要求7所述的抗体偶联物、如权利要求8所述的免疫细胞，或其组合；以及
(ii)药学上可接受的载体。
10.一种活性成分的用途，其特征在于，被用于(a)制备SARS-CoV2病毒感染的诊断试剂或试剂盒；和/或(b)制备预防和/或治疗SARS-CoV2病毒感染的药物；
其中，所述活性成分选自下组：如权利要求6所述的融合蛋白、如权利要求7所述的抗体偶联物、如权利要求8所述的免疫细胞，或其组合。
11.一种融合蛋白的制备方法，其特征在于，所述的融合蛋白的序列中包含：
(a)抗病毒(SARS、MERS或其他病毒)表面蛋白抗体；和
(b)机体内介导病毒侵染的蛋白(如SARS的ACE2、MERS的DPP4等)或其片段。
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