CN107438625A - 使用嵌合抗原受体（cars）的胎盘样硫酸软骨素的免疫治疗靶向和使用具有分割蛋白结合系统的cars的癌症的免疫治疗靶向 - Google Patents

使用嵌合抗原受体（cars）的胎盘样硫酸软骨素的免疫治疗靶向和使用具有分割蛋白结合系统的cars的癌症的免疫治疗靶向 Download PDF

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CN107438625A
CN107438625A CN201680018764.0A CN201680018764A CN107438625A CN 107438625 A CN107438625 A CN 107438625A CN 201680018764 A CN201680018764 A CN 201680018764A CN 107438625 A CN107438625 A CN 107438625A
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Abstract

本发明涉及包含有条件地有活性的嵌合抗原受体(CAR)和分割蛋白结合系统、VAR2CSA多肽或任何其它靶向剂的多肽。本发明还涉及包含VAR2CSA多肽或其它靶向剂以及分割蛋白结合系统的多肽。

Description

使用嵌合抗原受体（CARS）的胎盘样硫酸软骨素的免疫治疗靶 向和使用具有分割蛋白结合系统的CARS的癌症的免疫治疗 靶向
发明领域
本发明提供了组合物和方法，其用于使用与VAR2CSA融合的嵌合抗原受体(CAR)即VAR2-CAR或与分割蛋白结合系统融合的嵌合抗原受体(CAR)即Spy-CAR的癌症免疫疗法治疗，编码这些构建体的载体和包含CAR的重组T-细胞。
发明背景
在基于细胞的过继免疫治疗中，可以修饰从患者分离的免疫细胞以表达合成的蛋白质，其使得细胞在其随后转移回患者体内后能够执行新的治疗功能。这样的合成蛋白质的实例是嵌合抗原受体(CAR)。目前使用的CAR的实例是细胞外靶标识别结构域(例如，抗原结合 ScFv链)、跨膜结构域(例如来自CD8或CD28)和一个或多个细胞内信号传导结构域(包括CD28，OX40/4-1BB和CD3ζ)的融合物。抗原接合后，CAR的细胞内信号传导部分可以在免疫细胞中引发活化相关的反应；如溶细胞分子的释放以诱导肿瘤细胞死亡等。嵌合抗原受体(CAR)描述于Porter等人,2011,N.Engl.J.Med.365:725-733；和Kalos等人,2011,Sci.Transl.Med.3:95ra73中。
分割蛋白结合系统(SPBS)的特征在于蛋白质的两个分割片段重构成稳定的共价锁定结构的能力。这种SPBS的一个例子是 Spy-tag-Spy-Catcher系统，其包含来自化脓性链球菌(PMID： 20235501)的纤连蛋白结合蛋白(FbaB)的第二免疫球蛋白样胶原粘附素结构域(CnaB2)的反应性亚单位蛋白质伴侣(Spy-Tag和 Spy-Catcher)。SpyTag和SpyCatcher通过自发的多肽键形成相互作用，并且是分割蛋白结合系统的一个实例(Zakeri,B.等人PNAS. 2012)。如果一个蛋白质含有SpyTag，另一个蛋白质含有SpyCather，这两个蛋白质将通过SpyTag和SpyCather之间的共价相互作用自发连接，这确保了SpyTag和SpyCatcher连接蛋白之间的高结合强度和一对一的相互作用。Spytag-SpyCatcher系统与其它分割蛋白结合系统一起在Veggiani等人,2014,Trends Biotechnol.Oct；32(10):506-12 中描述。SpyCatcher蛋白质序列(116aa)在体内显示出一定程度的免疫原性，其可以通过截短蛋白质的N-末端部分(aa 1-24)(PMID： 25434527)来减小。虽然是非免疫原性的，但是ΔN-SpyCatcher序列保留了对13aa Spytag序列(PMID：25434527)的完全结合能力，因此可用于免疫敏感应用，如疫苗生产或嵌合免疫受体。
硫酸软骨素A(CSA)属于糖胺聚糖(GAG)家族，其是交替的氨基糖和己糖醛酸残基的连接于蛋白聚糖的线性聚合物。存在几种类型的GAG，包括：硫酸软骨素(CS)，硫酸皮肤素(DS或CSB)，硫酸乙酰肝素(HS)和肝素。虽然这些GAG的多糖主链是简单的，但在诸如硫酸化和糖醛酸差向异构化的修饰中出现相当大的多样性。这是不同GAG的域结构和生物活性的广泛多样性的基础。在包括癌症在内的各种人类疾病中已经报道了CSA的异常或不适当的表达。 CSA链被定义为主要是硫酸软骨素A，但CSA链也可以沿着CSA链具有少数DS或CSC。
疟原虫恶性疟原虫(Plasmodium falciparum)在其复杂生命周期的几乎所有阶段都使用宿主细胞蛋白聚糖。子孢子通过与高度硫酸化的硫酸乙酰肝素蛋白聚糖(HSPG)相互作用的表面表达的环子孢子蛋白感染肝脏中的肝细胞。红细胞的裂殖子感染是通过与血型糖蛋白 A上的唾液酸结合的EBA-175介导的。此外，许多介导宿主内皮粘附的恶性疟原虫红细胞膜蛋白1(PfEMP1)蛋白已被描述为聚糖结合性的。其中之一是VAR2CSA，它是PfEMP1蛋白家族的独特成员。 VAR2CSA以高亲和力结合胎盘的绒毛间隙中的附接至蛋白聚糖(所谓的硫酸软骨素蛋白聚糖(CSPG))的特定类型的硫酸软骨素A (CSA)。VAR2CSA是在恶性疟原虫感染的红细胞(IE)表面上表达的大的多结构域蛋白(350KDa)，并且VAR2CSA-CSA相互作用负责胎盘疟疾(PM)中的胎盘特异性隔离(placenta specific sequestration)。重要的是，来自FCR3和3D7型寄生虫的重组全长 VAR2CSA胞外结构域在低纳摩尔范围内显示出对CSA的亲和力。
发明目的
本发明的实施方案的目的是提供使用与VAR2CSA融合的嵌合抗原受体(CAR)即VAR2-CAR或与分割蛋白结合系统融合的嵌合抗原受体(CAR)即Spy-CAR的组合物和方法，编码其的载体，以及包含CAR的重组T细胞。在一些实施方案中，CAR的scFv部分被分割蛋白结合系统的一部分，例如SpyTag(SEQ ID NO：130)， SpyCatcher(SEQ ID NO：129)或SpyCatcher-ΔN(SEQ ID NO： 136)取代。在这些实施方案中，疾病抗原识别分子例如抗体，scFv 单链，纳米抗体或重组蛋白(包括单特异性或双特异性的VAR2CSA 肽或其片段)通过分割蛋白结合系统连接到CAR。本发明还涉及使用 VAR2CSA的硫酸软骨素A(CSA)修饰特异性的CAR。在一些实施方案中，CAR的scFv部分被至少包含最小CSA结合结构域ID1-ID2a 的VAR2CSA变体所取代。本发明的实施方案的其它目的是提供用于使用用于治疗与抗原例如疾病相关抗原例如疾病相关CSA的不适当表达相关的病况的能够结合分割蛋白的CAR的方法。本发明实施方案的其它目的是提供其中VAR2CSA嵌合构建体、其多肽或片段用于靶向具有CSA的表达例如不适当表达的组织或细胞的方法。本发明还包括能够植入针对细胞和组织的免疫应答的基于VAR2CSA的双特异性融合蛋白(双特异性性VAR2)，例如VAR2CSA和免疫细胞结合结构域之间的融合蛋白，例如scFv抗CD3结构域。
发明概述
本发明人已经发现重组VAR2CSA以高亲和力和特异性结合在疾病相关蛋白聚糖的有限库上的胎盘样CSA链。更重要的是，本发明人已经发现，胎盘样CSA链在几乎所有类型的人类肿瘤中独特地重新表达，并且重组VAR2CSA蛋白以高和特异性的亲和力结合癌细胞上和原发癌组织中的胎盘样CSA。VAR2SCA的此功能可用于提供适合于免疫治疗的构建体，该构建体包含CAR的元件或结构域，例如至少包含CD28和CD3ζ结构域。
因此，本发明人提示使用VAR2CSA和CSA之间的该特异性和高亲和力结合来植入针对具有胎盘样CSA修饰的蛋白聚糖的高表达或其它方式的不适当表达的人类肿瘤和转移的免疫应答。这可以通过 CAR和VAR2CSA之间的直接遗传融合(VAR2-CAR)获得。也可以通过在分割蛋白结合系统的片段(例如SpyTag或SpyCatcher)上融合到CAR(Spy-CAR)获得，并随后体外或体内附接融合到相应的 SpyTag或SpyCatcher的重组VAR2CSA蛋白。Spy-CAR可用于附接靶向分子不同于VAR2CSA(例如抗体或抗体片段)的其它情况。通过使用对胎盘样CSA和免疫细胞如CD3阳性细胞具有双特异性的双特异性VAR2CSA蛋白，也可以获得免疫细胞至具有胎盘样CSA的高表达或不适当表达的肿瘤组织或其它组织的植入。这通过将 VAR2CSA与特异于某一免疫细胞群体(如CD3阳性细胞)的单链抗体(如scFv抗CD3)融合获得。通过将CAR技术与分割蛋白质系统相结合，可以制备通用CAR，其可以通过用与分割蛋白的其它部分融合的癌靶向蛋白进一步治疗患者来进行个性化治疗。可以控制剂量，治疗可以停止，并且在治疗方案的过程中可以改变治疗。特别是通过制作分割系统CAR和相应的VAR2CSA分割蛋白，可以在治疗患者之前完美控制VAR2CSA蛋白质折叠、结合亲和力、结合特异性以及良好生产规范的二次修饰的监测。
因此，在第一方面，本发明涉及一种多肽，其包含i)第一多肽结构域，其是嵌合抗原受体(CAR)的跨膜结构域和胞内结构域，以及 ii)选自VAR2CSA多肽或分割蛋白结合系统的肽部分的第二胞外多肽结构域。
在第二方面，本发明涉及多肽，其包含i)第一多肽结构域，其是 VAR2CSA多肽或其它肿瘤靶向剂，例如单链scFv，和ii)选自分割蛋白结合系统的肽部分、单结构域抗体例如scFv或纳米抗体中的一个或多个的第二多肽结构域。
应当理解，本文所述的包含CAR和分割蛋白结合系统的肽部分的任何构建体可以以任何方向在嵌合构建体中正向或反向翻译形成。因此，作为实例，spytaq氨基酸序列可以是AHIVMVDAYKPTK(SEQ ID NO：130)或反向的KTPKYADVMVIHA(SEQ ID NO：137)。
在一些实施方案中，根据本发明的VAR2CSA的分离的蛋白质片段包含ID2a。
在第三方面，本发明涉及包含编码本发明多肽的核苷酸序列的分离的核酸分子。
在另一方面，本发明涉及包含分离的核酸分子的表达载体，其包含编码本发明多肽的核苷酸序列。
在另一方面，本发明涉及包含本发明载体的细胞。在一些实施方案中，细胞选自αβΤ细胞，γδT细胞，自然杀伤(NK)细胞，细胞毒性T淋巴细胞(CTL)，调节性T细胞或其任何组合。在另一方面，本发明涉及双组分治疗剂，其包含
a)第一组分，其是多肽，所述多肽包含i)第一多肽结构域，其是嵌合抗原受体(CAR)的跨膜结构域和胞内结构域，以及ii)第二多肽结构域，其是连接至所述CAR的胞外部分的分割蛋白结合系统的肽部分；和
b)第二组分，其是多肽，所述多肽包含i)第一多肽结构域，其是VAR2CSA多肽或任何其它肿瘤靶向剂，例如抗体或scFv，以及ii) 第二多肽结构域，其是分割蛋白结合系统的肽部分；任选地还包含iii) 单结构域抗体，例如scFv或纳米抗体，
其中由组分a)和b)定义的分割蛋白结合系统的所述肽部分自发形成不可逆的异肽键。
在另一方面，本发明涉及治疗或预防受试者中的与例如癌症中 CSA的表达例如不适当表达相关的任何适应症或病况的发生的方法，所述方法包括：(a)在促进活化的条件下培养细胞毒性淋巴细胞群； (b)用编码多肽的载体转染(a)的淋巴细胞群，所述多肽包含i)第一多肽结构域，其是嵌合抗原受体(CAR)的跨膜结构域和胞内结构域，以及ii)第二多肽结构域，其选自VAR2CSA多肽或任何其它靶向剂，例如抗体；(c)将治疗或预防有效量的(b)的经转染的淋巴细胞施用于具有所述适应症或处于具有所述适应症的风险中的受试者；其中包含CAR的结构域的多肽具有针对靶向部分的结合特异性，或可被靶向部分结合；从而治疗或预防所述受试者的所述适应症。
在另一方面，本发明涉及治疗或预防受试者中的与例如癌症中 CSA的表达例如不适当表达相关的任何适应症或病况的发生的方法，所述方法包括：(a)在促进活化的条件下培养细胞毒性淋巴细胞群； (b)用编码多肽的载体转染(a)的淋巴细胞群，所述多肽包含i)第一多肽结构域，其是嵌合抗原受体(CAR)的跨膜结构域和胞内结构域，以及ii)第二多肽结构域，其是连接至所述CAR的胞外部分的分割蛋白结合系统的肽部分；(c)将治疗或预防有效量的(b)的经转染的淋巴细胞施用于具有所述适应症或处于具有所述适应症的风险中的受试者；和(d)施用多肽，所述多肽包含i)第一多肽结构域，其为VAR2CSA多肽或其它靶向剂，例如抗体，和ii)第二多肽结构域，其是分割蛋白结合系统的肽部分，其中在两种不同的多肽中定义的分割蛋白结合系统的所述肽部分自发形成不可逆的异肽键；从而治疗或预防所述受试者的所述适应症。
在另一方面，本发明涉及用作药物的根据本发明的多肽或根据本发明的双组分治疗剂。
在另一方面，本发明涉及包含根据本发明的多肽或根据本发明的双组分治疗剂的药物组合物。
在另一方面，本发明涉及根据本发明的多肽或根据本发明的双组分治疗剂用于制备药物的用途。
在另一方面，本发明涉及根据本发明的多肽或根据本发明的双组分治疗剂或根据本发明的药物组合物，用于治疗例如癌症，关节炎，关节病，多发性硬化和神经损伤后愈合，软骨修复，伤口愈合和牛皮癣中与涉及CSA的表达例如不适当表达的病况相关的任何适应症。
在另一方面，本发明涉及与任何其它合适的抗癌剂的组合的根据本发明的多肽，或根据本发明的双组分治疗剂或根据本发明的药物组合物。
在一些实施方案中，癌症选自所有表达CSA的恶性肿瘤，包括癌 (包括但不限于乳腺癌，胰腺癌，卵巢癌，子宫内膜癌，肝细胞癌，肺癌，结肠癌，前列腺癌，宫颈癌，睾丸癌，基底细胞皮肤癌，透明细胞肾细胞癌，头颈鳞状细胞癌，皮肤鳞状细胞癌，外阴角质化鳞状细胞癌和外阴基底细胞癌)，肉瘤(包括但不限于纤维肉瘤，去分化的软骨和脂肪肉瘤，平滑肌肉瘤，脂肪肉瘤，粘液样脂肪肉瘤，子宫体平滑肌肉瘤，骨肉瘤，尤文肉瘤和横纹肌肉瘤，滑膜肉瘤，孤立性纤维瘤)，造血系统癌(包括但不限于慢性淋巴细胞白血病(CLL)，急性淋巴细胞白血病(ALL)，急性骨髓性白血病(AML)，B细胞、 T细胞和大颗粒淋巴瘤)，神经上皮组织的肿瘤，例如但不限于星形细胞瘤(多形性黄色星形细胞瘤，纤维性星形细胞瘤，间变性星形细胞瘤，多形性成胶质细胞瘤)，少突神经胶质瘤，室管膜瘤，脉络丛肿瘤，少突星形细胞瘤，神经胶质肉瘤，神经节胶质细胞瘤，视网膜母细胞瘤，神经细胞瘤，神经母细胞瘤(成感觉神经细胞瘤和神经节成神经细胞瘤)，髓母细胞瘤，非典型畸胎样横纹肌样瘤和所有类型的神经内分泌癌以及任何其它表达CSA的癌症。
应当理解，对于包含VAR2CSA多肽的融合或嵌合蛋白，可以使用本文定义的任何VAR2CSA序列和多肽。因此，该方面不限于使用最小结合片段。这在使用术语VAR2CSA序列或多肽时均适用，并且在用于医学治疗或靶向时相应地是等同相关的。
发明详述
在一些实施方案中，本发明的多肽包含CAR的scFv部分和 VAR2CSA肽。在其它实施方案中，重组VAR2CSA肽使用分割-蛋白结合系统连接至CAR。在其它实施方案中，使用分割蛋白结合系统将疾病靶向单位(例如抗体，单链结合物(scFv)或重组肽)连接至CAR。本发明还包括基于VAR2CSA的融合蛋白，其能够通过双重结合至胎盘样CSA和免疫细胞特异性受体来募集针对表达胎盘样CSA的细胞和组织的免疫应答(双特异性VAR2)。
单或双特异性VAR2-CAR：
根据本发明的多肽构建体可以包含与CAR的跨膜结构域和胞内结构域组合的VAR2CSA。在本发明的一些方面，修饰宿主的免疫细胞以表达本发明的包含CAR的所述多肽构建体。人类T淋巴细胞表达肿瘤定向嵌合抗原受体(CAR)的遗传工程改造可以产生抗肿瘤的效应细胞，其能够绕过肿瘤免疫逃逸机制，这是由于蛋白质-抗原加工和呈递中的异常。此外，这些转基因受体可以针对不是蛋白质来源的肿瘤相关抗原。在本公开内容的某些实施方案中，存在被修饰以包含至少一种CAR的CTL。
在特定情况下，细胞毒性T淋巴细胞(CTLs)包含嵌合、非天然和至少部分地手工改造的受体。在特定情况下，除了CAR之外，根据本发明的多肽构建体还包含一种，两种，三种，四种或更多种组分，并且在一些实施方案中，一种或多种组分促进T淋巴细胞靶向或结合到包含肿瘤抗原(例如VAR2CSA多肽)的癌细胞。在具体实施方案中，根据本发明的多肽构建体包含作为胞质信号传导结构域的部分或全部的和/或一种或多种共刺激分子的一部分或全部(例如共刺激分子的胞内结构域)的CAR，和单独的(单特异性)VAR2CSA多肽或与特异于肿瘤抗原的抗体、单链scFV或重组肽融合的VAR2CSA 多肽(双特异性VAR2-CAR)。在某些方面，抗体针对可由例如替莫唑胺(TMZ)治疗的癌细胞的细胞表面上的靶抗原。在某些实施方案中，使用胞质信号传导结构域，例如衍生自T细胞受体ζ-链的那些，作为嵌合受体至少一部分，以便在嵌合受体与目标抗原接合之后产生 T淋巴细胞增殖的刺激信号和效应子功能。实例将包括但不限于来自共刺激分子如CD27，CD28，4-IBB和OX40的胞内结构域或细胞因子受体如IL7和IL15的信号传导成分。在特定实施方案中，共刺激分子用于增强抗原接合后由CAR产生的T细胞的活化、增殖和细胞毒性。在具体实施方案中，共刺激分子是CD28，OX40和4-1BB，细胞因子和细胞因子受体是IL7和IL15。
CAR可以是第一代，第二代或第三代(其中信号传导由CD3ζ以及由CD28和肿瘤坏死因子受体(TNFr)结构域，例如4-1BB或OX40 提供的共刺激提供的CAR)或任何其它CAR，其包含至少一个这些结构域。根据本发明的VAR2-CAR的多肽构建体还可以含有信号肽 (SP)，使得构建体的CAR部分可以被糖基化并锚定在免疫效应细胞的细胞膜中。跨膜结构域(TD)将胞外结构域连接到胞内结构域并当由细胞表达时驻留在细胞膜内，并且可以衍生自天然或合成来源。如果来源是天然的，则该结构域可以衍生自任何膜结合或跨膜蛋白。例如，跨膜区域可以衍生自T细胞受体，CD28，CD3ε，CD45，CD4， CD5，CD8，CD9，CD16，CD22，CD33，CD37，CD64，CD80，CD86， CD134，CD137或CD154的α、β或ζ链(即，至少包含其跨膜区域)。或者，TD可以是合成的，在这种情况下，它将主要包含疏水残基，如亮氨酸和缬氨酸。在一些情况下，在合成的TD的每个末端将发现苯丙氨酸、色氨酸和缬氨酸的三元组。
CAR还可以包括铰链序列。铰链序列是促进蛋白质柔性的氨基酸的短序列(参见，例如，Woof等人,Nat.Rev.Immunol,4(2):89-99 (2004))。铰链序列可以位于抗原识别部分和TD之间。铰链序列可以是从任何合适的分子衍生或获得的任何合适的序列。在一些实施方案中，例如，铰链序列衍生自CD8a分子或CD28分子，或衍生自 IgG，例如IgG1铰链区
根据本发明的包含VAR2-CAR的多肽构建体可以针对其它疾病相关表位具有双重特异性，包括但不限于CD19，CD20，CD22，κ或轻链，CD30，CD33，CD123，CD38，ROR1，HER2/ErbB2，ErbB3/4， EGFR，EGFRvIII，EphA2，FAP，癌胚抗原，EGP2，EGP40，间皮素，TAG72，PSMA，NKG2D配体，B7-H6，IL-13受体cc2，IL-11 受体a，MUC1，MUC16，CA9，GD2，GD3，HMW-MAA，CD171， Lewis Y，G250/CAIX，HLA-AI MAGE Al，HLA-A2NY-ESO-1，PSC1，叶酸受体α，CD44v7/8，8H9，NCAM，VEGF受体，5T4，胎儿AchR， NKG2D配体，CD44v6，TEM1，TEM8，由肿瘤表达的病毒相关抗原或通过基因组分析和肿瘤差异表达研究鉴定的其它肿瘤相关抗原。
根据本发明的包含嵌合抗原受体(CAR)的跨膜结构域和胞内结构域的多肽构建体可以进一步包含其它调节元件或自杀基因系统，例如诱导型半胱天冬酶-9(iCASP9)(Clackson T.等人(1998).Proc.Natl. Acad.Sci.U.S.A.95 10437–10442；StraathofK.C.等人(2005). Blood 105 4247–4254),或睡美人(SB)转座子系统(Maiti SN等人,JImmunother.2013Feb；36(2):112-23)。
分割蛋白结合系统(SPBS)：
在本发明中，SpyTag-SpyCatcher SPBS可以与CAR技术组合。用SPBS的片段(例如SpyCatcher序列)替代CAR构建体的scFv 结构域将允许与SPBS的另一部分融合的靶向单元(例如scFv和抗体或重组蛋白)的翻译后连接到CAR。SpyTag和SpyCatcher之间的相互作用在Lys31的未质子化的胺亲核攻击Asp117的羰基碳时发生，由邻近的Glu77催化。介导这种结合的最小肽是AHIVMVDA(SEQ ID NO：131)，而产生序列AHIVMVDAYKPTK(SEQ ID NO：130)的c-末端延伸提供了称为“SpyTag”的最佳区域(Zakeri等人PNAS 2012)。
SpyCatcher是来自化脓性链球菌的FbaB蛋白的CnaB2结构域的一部分，并且与由CnaB2结构域的另一部分组成的SpyTag结合。当 CnaB2结构域的这两个多肽混合时，它们会自发形成不可逆的异肽键，从而完成CnaB2结构域的形成。
在本发明中也可以使用K-Tag/SpyTag/SpyLigase系统。来自化脓链球菌的CnaB2结构域可用于产生共价肽-肽连接系统(Fierer JO等人，2014)。这通过将CnaB2分割为三个部分来完成：a)13个氨基酸的SpyTag(SEQ ID NO：130)，b)称为K-Tag的CnaB2的β- 链(SEQID NO 132)，和c)由其余的SpyCatcher多肽构建的 SpyLigase(SEQ ID NO：133)。通过表达具有在C-或N-末端融合的小K-Tag的靶向蛋白质，并将该融合蛋白与呈现SpyTag的T细胞以及SpyLigase一起混合，K-Tag融合抗原将通过SpyLigase促进的SpyTag与K-Tag的共价连接附接至SpyTag-VLP。相反，K-Tag也可以遗传地插入到CAR中，由此靶向抗原随后应在C-或N-末端被融合到SpyTag。
作为在T细胞表面上靶向抗原的共价偶联的相似策略的一部分，另外一对分割蛋白结合配偶体可用于本发明。来自化脓性链球菌的主要菌毛蛋白Spy0128可以被分割成两个片段split-Spy0128(Spy0128 的残基18-299)(SEQ ID NO：135)和异肽(Spy0128的残基293-308 (TDKDMTITFTNKKDAE))(SEQ ID NO 134)，它们共同形成分子间共价络合物(Zakeri，B.等人，2010)。与描述的 SpyTag-SpyCatcher策略一致，可以将Spy0128异肽插入到TCR或靶向抗原中，并且对于split-Spy0128结合配偶体也是相同的。再次简单混合或预注射标记的靶向蛋白将导致T细胞上标记的CAR与靶向抗原之间的共价相互作用。
Spy-CAR也可用于通过将K-标记的抗体，K-tag连接的抗体， K-tag连接的scFv链，K-标记的重组蛋白或其组合连接到Spy-CAR 来构建双特异性或多特异性CAR，获得双特异性或多特异性CAR构建体。
应该理解，SpyTag可能被Spy0128异肽取代，SpyCatcher由 split-Spy0128取代。
目前使用的CAR不易于药理学控制。分割蛋白结合系统的使用可以促进CAR能够在体内针对特定抗原激活和失活。
本发明的一个重要方面是i)收集，ii)修饰和iii)输注来自具有高表达或不适当表达的胎盘样CSA的受试者(例如癌症患者)的T 细胞群。在收集步骤中，从患者血液样品中纯化T细胞。在修饰步骤中，用编码由至少胞外分割蛋白结合结构域(如SpyTag或SpyCatcher)、跨膜结构域(如CD8或CD28)和激活结构域(如 CD3ζ)组成的CAR的载体转染T细胞群。然后静脉内施用与分割蛋白结合系统的其它部分标记、缀合或融合的抗原结合分子，例如抗体，单链scFv或重组VAR2CSA多肽。该分子将结合由CAR提供的分割蛋白的相应部分，这将使得CAR能够结合感兴趣的靶标并激活。在输注步骤中，将表达CAR的重组T细胞注射到患者体内，建立能够对靶标反应的T细胞群。
该原理可以应用于任何肿瘤结合分子(包括但不限于抗体，scFv 单链和重组蛋白)，并且具有如下优点：
a)可以用相同的CAR输注靶向许多不同的肿瘤抗原或靶标。
b)可以通过暂停输注spyTag(或类似的肽)肿瘤靶标分子来恢复治疗。
c)可以使用复合靶向剂，例如重组VAR2SCA，其不容易制成嵌合体作为CAR构建体的一部分。
双特异性VAR2蛋白：
由于VARCSA可以作为单链重组蛋白产生，因此可以与能够识别特定疾病靶标的其它单链蛋白融合。例如，将VAR2CSA融合到靶向CD3的scFv链将能够将表达CD3的免疫细胞募集到表达胎盘样 CSA的细胞和组织。因此，这种双重特异性的基于VAR2CSA的分子可用于植入针对具有胎盘样CSA表达的细胞和组织的T细胞应答。在一些实施方案中，VAR2CSA可以遗传融合到单链抗体，例如scFv 链或纳米抗体，产生双特异性VAR2CSA蛋白。在一些实施方案中，这种双特异性VAR2CSA蛋白可以含有分割蛋白结合标签，例如 SpyTag或K-Tag。在一些实施方案中，双特异性VAR2CSA蛋白可以通过分裂结合蛋白系统连接到CAR。
VAR2CSA多肽
本发明基于以下事实：疟疾蛋白的一部分(所谓的VAR2CSA) 可以以非常高的特异性和非常高的结合强度结合癌症特异性CSA和细胞外CSA缀合的CSPG。该部分技术描述于WO2013117705A1中。
CSA与许多重要因素如生长激素，细胞因子，趋化因子和粘附分子相互作用，并被认为参与结构稳定，胞质分裂，细胞增殖，分化，细胞迁移，组织形态发生，器官发生，感染和伤口修复。CS链由N- 乙酰基-D-半乳糖胺(GalNAc)和葡萄糖醛酸残基的交替单元组成。葡萄糖醛酸可以在其C2位硫酸化，GalNAc可以在C4和/或C6硫酸化，产生各种二糖单元。糖骨架的不同修饰允许CS链的结构和功能异质性。胎盘粘附恶性疟原虫寄生虫与主要为C4硫酸化的CS链特异性结合。
已经显示重组VAR2CSA蛋白以前所未有的高亲和力和特异性结合CSA。这可能是由于与CSA的相互作用，其不仅取决于带电的硫酸盐，而且还取决于CS主链。本发明的发明人已经证明，存在于胎盘中的CSA非常类似于在各种癌细胞包括癌症干细胞上呈现的CSA。这通过表达VAR2CSA的疟疾寄生虫特异性地粘附至C32黑素瘤细胞上的CSA和人类脑癌细胞的事实得到证明。
因此，本发明依赖于VAR2CSA重组蛋白和胎盘样CSA之间的高亲和性和特异性。通过标记该蛋白质，本发明可用于广泛的应用，包括在体内追踪转移瘤和诊断转移性疾病。通过将VAR2CSA偶联到凋亡或细胞毒性试剂上，本发明可用于特异性靶向和消除癌细胞和癌症干细胞。通过使用VAR2CSA重组蛋白的简单治疗，可以中和CSA 的活性，从而抑制表达CSA的癌细胞的肿瘤发生和/或转移。CSA可以存在于许多蛋白质主链上，例如CSPG4，CD44，双糖聚糖，核心蛋白聚糖，多能聚糖，聚集蛋白聚糖(软骨中的主要CSPG)，基底膜聚糖，多配体聚糖。
本发明被设想为与恶性黑素瘤癌(包括在黑素瘤细胞表面具有 CSA链的具有CSPG4的皮肤、眼和结膜黑素瘤)特别相关。该GAG 链被认为参与有丝分裂和转移。然而，CSPG4不仅是特异于黑素瘤。本发明人对来自人体组织和细胞系的大实验组的数据进行的微阵列和组织阵列分析表明CSPG4和其它类型的含CSA的蛋白聚糖可能存在于源自所有三种细胞胚层的广泛范围的癌症类型。这些癌症类型包括癌(乳腺癌，胰腺癌，卵巢癌，子宫内膜癌，肝细胞癌，肺癌，结肠癌，前列腺癌，宫颈癌，睾丸癌，基底细胞皮肤癌，透明细胞肾细胞癌，头颈鳞状细胞癌，皮肤鳞状细胞癌，外阴角质化鳞状细胞癌和外阴基底细胞癌)，肉瘤(乳腺脂肪肉瘤，纤维肉瘤，去分化的软骨和脂肪肉瘤，平滑肌肉瘤，脂肪肉瘤，粘液样脂肪肉瘤，子宫体平滑肌肉瘤，骨肉瘤，尤文肉瘤和横纹肌肉瘤)，造血系统癌(慢性淋巴细胞白血病(CLL)，急性淋巴细胞白血病(ALL)，急性骨髓性白血病(AML)，B细胞、T细胞和大颗粒淋巴瘤)，神经上皮组织的肿瘤，例如星形细胞瘤(多形性黄色星形细胞瘤，纤维性星形细胞瘤，间变性星形细胞瘤，多形性成胶质细胞瘤)，少突神经胶质瘤，室管膜瘤，脉络丛肿瘤，少突星形细胞瘤，神经胶质肉瘤，神经节胶质细胞瘤，视网膜母细胞瘤，神经细胞瘤，神经母细胞瘤(成感觉神经细胞瘤和神经节成神经细胞瘤)，髓母细胞瘤，非典型畸胎样横纹肌样瘤和所有类型的神经内分泌癌。
本发明的发明人已经鉴定了一种疟疾蛋白VAR2CSA，其以小于10nM的亲和力结合胎盘的间隙空间中的CSA。已经以高产率生产了 VAR2CSA的较小的重组部分，其结合具有与全长和天然VAR2CSA 蛋白质相似的特征的CSA。重组VAR2CSA蛋白不与其它类型的CS (例如硫酸软骨素C(C6S)或高度硫酸化的GAG如硫酸乙酰肝素 (HS))结合。可以产生重组蛋白质以在各种表达系统(包括S2细胞，T.Ni细胞，CHO细胞和大肠杆菌菌株包括BL21和Shuffle细胞 (tm New England Biolabs))中以高亲和力结合CSA。
本发明的发明人还鉴定了以非常高的亲和力(nM)和高特异性结合CSA的单个小(75KDa)抗原。表3(参见实施例2)列出了使用生物传感器技术的分析的所有VAR2CSA蛋白的CSA亲和力。
本发明的发明人已经表明，该VAR2CSA重组蛋白以低浓度强烈地结合到广泛的癌细胞系，包括皮肤黑素瘤(C32，MeWo)，肺癌 (A549)，乳腺癌(HCC1395)，骨肉瘤(U2OS，MNNG/HOS)，横纹肌肉瘤(RH30)和皮肤T细胞淋巴瘤(表4和5)。作为对照分子，使用另外一种VAR2CSA蛋白，其与最小结合性VAR2CSA构建体相同，除了在分子的C-末端部分中151个氨基酸的截短。此截短移除CSA结合。重组VAR2CSA结合所有表达CSPG4的细胞系和表达具有CSA链的其它CSPG分子的癌细胞系(例如T细胞淋巴瘤)。重组VAR2CSA蛋白不能与人红细胞和外周血单核细胞(PBMC)相互作用(表4)。
本发明的发明人已经通过VAR2CSA与附接于给定的蛋白聚糖的胎盘样CSA链之间的特异性相互作用，显示疟疾寄生虫粘附于癌细胞和原发性人类肿瘤。因此，可以设想重组VAR2CSA及其缀合物可用于引导针对肿瘤环境的免疫应答。
使用VAR2CSA靶向肿瘤上的CSA相对于其它现有的治疗方法的优势有很多：
1)VAR2CSA和CSA之间的相互作用具有前所未有的高亲和力和高度特异性。
2)VAR2CSA是进化型精制疟疾蛋白，因此治疗不太可能会破坏患者的耐受性并引起自身免疫反应。
3)VAR2CSA是稳定的蛋白质，具有很好的表征，可以在与大规模蛋白质生产相容的生物体中高度表达。
4)VAR2CSA是具有多种血清变体的多态性蛋白质。反复治疗可通过不同的血清变体提供，以避免中和抗体的问题。
5)VAR2CSA天然暴露在恶性疟原虫感染的红细胞上，因此本质上是人血清中稳定的蛋白质，并且已被证明是高度抗蛋白酶的。
使用双特异性的基于VAR2CSA的分子(如VAR2CSA-scFv-CD3 融合蛋白)的优点众多：
1)VAR2CSA结合由多种恶性疾病适应症表达的常见的与癌症相关的CSA修饰，因此双特异性VAR2SCA融合蛋白可以将T细胞应答直接植入CSA阳性肿瘤环境。
2)由于在给定的恶性疾病中表达的几种蛋白聚糖上存在恶性相关的CSA修饰，所以给定的癌症不太可能通过下调特定的蛋白聚糖来产生对双特异性VAR2CSA融合蛋白的抗性。
3)由于VAR2CSA序列在疟原虫恶性疟原虫的进化过程中被天然优化，以逃避人类免疫防御机制和血清蛋白酶，因此患者不太可能发展针对双特异性VAR2CSA-融合蛋白的免疫(或其它)抗性。
4)表达VAR2CSA的疟原虫只能隔离到胎盘(并且不能在人体/ 脉管系统中其它地方发现)的事实证明，双特异性VAR2CSA融合蛋白将被预测在治疗环境中具有最小的不利影响。
使用VAR2CSA-CAR相对于基于scFv的CAR的优点有很多：
1)VAR2CSA结合由多种恶性疾病适应症表达的常见癌症相关 CSA修饰，因此VAR2-CAR具有广泛但癌症特异性的靶向范围，并且可用于多种癌症适应症。
2)由于在给定的恶性疾病中表达的几种蛋白聚糖中存在恶性相关的CSA修饰，所以给定的癌症不可能通过下调特异性的蛋白聚糖来产生对VAR2CSA-CAR的抗性。
3)由于VAR2CSA序列在恶性疟原虫疟原虫的进化过程中被自然优化，以逃避人类免疫防御机制和血清蛋白酶，因此患者不太可能针对基于VAR2CSA的CAR发展免疫(或其它)抗性。
4)表达VAR2CSA的疟原虫只能隔离到胎盘(并且不能在人体/ 脉管系统中其它地方发现)的事实证明，VAR2CSA-CAR将被预测在治疗环境中具有最小的不利影响。
使用分割蛋白结合系统(Spy-CAR)的CAR相对于传统的第一代，第二代和第三代CAR的优点有很多：
1)Spy-CAR本身不构成靶向序列，本身不会对人抗原产生反应。
2)Spy-CAR可以使用VAR2CSA-Spy蛋白“武装”，这将使 VAR2CSA-Spy-Spy-CAR能够针对所选择的疾病靶标产生反应。
3)Spy-CAR可以用任何给定的靶向分子进行“武装”，所述靶向分子与分割蛋白结合系统的相应片段融合或缀合，这将使靶向序列 -Spy-Spy-CAR能够针对所选择的疾病靶标产生反应。
4)Spy-CAR转导的T细胞可以在体外用VAR2CSA-Spy蛋白进行“武装”，这将使得VAR2CSA-Spy-Spy-CAR能够针对所选择的疾病靶标产生反应。
5)Spy-CAR转导的T细胞可以用任何给定的靶向分子进行“武装”，所述靶向分子在体外与融合蛋白结合系统的相应片段融合或缀合，这将使靶向序列-Spy-Spy-CAR能够针对所选择的疾病靶标产生反应。
6)Spy-CAR转导的T细胞可以用在体外通过K-tag彼此连接的靶向分子的任何给定的聚合物复合物进行“武装”，这将使得靶向序列 -Spy-Spy-CAR能够针对所选择的疾病靶标产生反应。
7)Spy-CAR转导的T细胞可以通过用与分割蛋白结合系统的相应片段融合或缀合的靶向序列追踪Spy-CAR转导的T细胞而在体内用VAR2CSA-Spy蛋白进行“武装”，这将使得VAR2CSA-Spy-Spy-CAR能够能够针对所选择的疾病靶标产生反应。
8)Spy-CAR转导的T细胞可以通过用与分割蛋白结合系统的相应片段融合或缀合的靶向序列追踪Spy-CAR转导的T细胞而在体内用与分割蛋白结合系统的相应片段融合或缀合的任何给定的靶向分子进行“武装”，这将使得靶向序列-Spy-Spy-CAR能够能够针对所选择的疾病靶标产生反应。
9)Spy-CAR转导的T细胞可以通过用与分割蛋白结合系统的相应片段融合或缀合的靶向序列追踪Spy-CAR转导的T细胞而在体外或体内用通过K-tag彼此连接的靶向分子的任何给定聚合复合物进行“武装”，这将使得靶向序列-Spy-Spy-CAR能够能够针对所选择的疾病靶标产生反应。
10)由于活化的细胞毒性T细胞的寿命有限，VAR2CSA-Spy-Spy-CAR转导的T细胞或用与分割蛋白结合系统的相应片段融合或缀合的任何给定的靶向分子武装的T细胞将随着时间的推移从生物体中清除，同时保持“非武装”的表达Spy-CAR的记忆T 细胞。因此，VAR2CSA-Spy-Spy-CAR“武装”的T细胞群体将随着时间的推移通过在脾脏中自然清除而消除。在发生疾病复发的情况下，仅表达Spy-CAR的持续记忆性T细胞随后可以通过用与分割蛋白结合系统的相应片段融合的相同或新的疾病靶向序列追踪该细胞进行体内“重新武装”。
11)Spy-CAR可以用不能在重组T细胞中作为嵌合蛋白产生任何的癌靶向部分进行武装。
12)在CAR治疗期间，Spy-CAR可以首先用一个Spy-标记的蛋白质进行“武装”，并且在发生抗药性时，或者如果癌症改变表型，或者诱导对Spy-标记蛋白的免疫应答，则Spy-CAR可以用另一个二线 Spy-标记的癌症靶向分子武装。
定义
术语“SpyTag”是指来自化脓链球菌的FbaB蛋白质的CnaB2 结构域的一部分，其被优化以结合由CnaB2结构域的另一部分(SEQ ID NO：130和131)组成的SpyCatcher。当Lys31的未质子化的胺亲核攻击Asp117的羰基碳时发生相互作用，由相邻的Glu77催化。介导该结合的最小肽是AHIVMVDA，而产生序列 AHIVMVDAYKPTK的c末端延伸提供了称为“SpyTag”的最佳区域 (Zakeri,B.等人PNAS.2012)。
术语“SpyCatcher”是指来自化脓链球菌的FbaB蛋白质的CnaB2 结构域的一部分，其被优化为结合由CnaB2结构域的另一部分(SEQ ID NO：129)组成的SpyTag。SpyCatcher可以是CnaB2的残基1-113，并且结合可以通过以下两个突变进行优化：I34E和M69Y(Zakeri,B. 等人PNAS,2012)。
本文中可互换使用的术语“多核苷酸”和“核酸”是指任何长度的核苷酸(核糖核苷酸或脱氧核糖核苷酸)的聚合形式。因此，该术语包括但不限于单链，双链或多链DNA或RNA，基因组DNA，cDNA， DNA-RNA杂交体，或包含嘌呤和嘧啶碱基或其它天然、化学或生物化学修饰的、非天然的或衍生的核苷酸碱基的聚合物。
术语“抗体”和“免疫球蛋白”包括任何同种型的抗体或免疫球蛋白，保留与抗原特异性结合的抗体片段，包括但不限于Fab，Fv， scFv和Fd片段，嵌合抗体，人源化抗体，单链抗体和包含抗体的抗原结合部分和非抗体蛋白质的融合蛋白。
“抗体片段”包含完整抗体的一部分，例如完整抗体的抗原结合或可变区。抗体片段的实例包括Fab，Fab'，F(ab')2和Fv片段；双体；线性抗体(Zapata等人,Protein Eng.8(10):1057-1062(1995))；单链抗体分子；和由抗体片段形成的多特异性抗体。抗体的木瓜蛋白酶消化产生两个相同的抗原结合片段，称为“Fab”片段，各自具有单个抗原结合位点，以及残留的“Fc”片段，此名称反映了容易结晶的能力。
本文所用的术语“VAR2CSA多肽”是指由恶性疟原虫表达的特异性红细胞膜蛋白1(PfEMP1)蛋白的胞外部分，其与硫酸软骨素蛋白聚糖(CSPG)相互作用并且特征在于具有SEQ ID NO：55或SEQ ID NO：56或其片段或变体的序列，其具有结合可以呈现在蛋白聚糖(CSPG)上的硫酸软骨素A(CSA)的能力。
在一些实施方案中，根据本发明的VAR2CSA多肽至少包含 VAR2CSA的蛋白质片段，该片段由a)ID1和b)DBL2Xb的连续氨基酸序列组成。
在一些实施方案中，根据本发明的VAR2CSA多肽至少包含 VAR2CSA的蛋白质片段，该片段由a)ID1和b)DBL2Xb和c)ID2a 的连续氨基酸序列组成。
在VAR2CSA多肽的定义中包括在Salanti A.等人Mol.Micro 2003Jul；49(1):179-91,Khunrae P.等人,J Mol Biol.2010Apr 2；397(3):826-34,Srivastava A.等人,ProcNatl Acad Sci U S A.2010 Mar 16；107(11):4884-9, M.等人,J BiolChem.2011May 6；286(18):15908-17,或Srivastava A.等人,PLoS One. 2011；6(5):e20270中描述的多肽。
本文所用的术语“ID1”是指VAR2CSA的结构域，其特征在于具有与SEQ ID NO：1的1-152所鉴定的氨基酸序列具有至少70％序列同一性的氨基酸序列。
本文所用的术语“DBL2Xb”是指VAR2CSA的结构域，其特征在于具有与SEQ ID NO：1的153-577所鉴定的氨基酸序列具有至少70％序列同一性的氨基酸序列。
本文所用的术语“ID2a”是指VAR2CSA的结构域，其特征在于其氨基酸序列包含来自SEQ ID NO：1的氨基酸578-640的N-末端的至少20，至少21，至少22，至少23，至少24，至少25，至少26，至少27，至少28，至少29，至少30，至少31，至少32，至少33，至少34，至少35，至少36，至少37，至少38，至少39，至少40，至少41，至少42，至少43，至少44，至少45，至少46，至少47，至少48，至少49，至少50，至少51，至少52，至少53，至少54，至少55，至少56，至少57，至少58，至少59，至少60，至少61，或至少62，例如63个连续氨基酸，并且与这样的连续氨基酸序列具有至少70％的序列同一性。
在一些实施方案中，本文所指的由SEQ ID NO：1-75或其片段鉴定的任何一个序列的至少70％的氨基酸序列同一性是指与该序列具有至少71,72,73,74,75,76,77,78,79,80,81,8,83,84,85,86,87,88, 89,90,91,92,93,94,95,96,97,98或99％序列同一性的序列。
如本文所用的术语“变体”或“多个变体”是指具有SEQ ID NO：55或SEQ ID NO：56的氨基酸序列的VAR2CSA多肽或包含SEQ ID NO：1-54的氨基酸序列的片段VAR2CSA多肽，其片段或变体保留结合蛋白聚糖(CSPG)上的硫酸软骨素A(CSA)的能力，其中一个或多个氨基酸已经被另一个氨基酸取代和/或其中一个或多个氨基酸已缺失和/或其中一个或多个氨基酸已被插入到多肽中和/或其中一个或多个氨基酸已加入到该多肽中。这种添加可以在N-末端或C-末端或两者上发生。该定义中的“变体”或“多个变体”在能够结合硫酸软骨素A(CSA)方面仍具有功能活性。在一些实施方案中，变体与 SEQ ID NO：1-75和128的序列，例如SEQ ID NO：1,3-5,10,11,29, 34,36-38,41,43-45,48,53-56,60-70,72-75和128的序列具有至少 70％，例如至少71,72,73,74,75,76,77,78,79,80,81,82,83,84,85, 86,87,88,89,90,91,92,93,94,95,96,97,98或99％的序列同一性。
本文所用的术语“功能性变体”，“功能片段”和“功能性衍生物”是指SEQ ID NO：55或SEQ ID NO：56的变体，片段，截短形式以及衍生物，例如SEQ ID NO：1,3-5,10,11,29,34,36-38,41,43-45, 48,53-56,60-70,72-75和128的任一个，所述多肽包括SEQ ID NO：55或SEQ ID NO：56的必需的结合序列部分，并且至少具有结合硫酸软骨素A(CSA)的能力。应当理解，VAR2CSA功能变体或功能片段可以具有选自作为变体和/或片段和/或衍生物的两个或三个特征。
VAR2CSA多肽的功能性变体或片段包括当在本文所述的测定中测试时显示在相同细胞类型中产生的野生型VAR2CSA多肽的结合亲和力的至少约25％，例如至少约50％，例如至少约75％，例如至少约90％的那些。
本文所用的术语“免疫学片段”是指具有基本上相同的功能活性和以相同空间取向被抗体识别的氨基酸序列的片段。因此，特异性抗体将结合多肽和其免疫学片段。
本文所用的术语“另一个氨基酸”是指与该位置天然存在的氨基酸不同的一个氨基酸。这包括但不限于可由多核苷酸编码的氨基酸。在一些实施方案中，不同的氨基酸是天然的L-型并且可以由多核苷酸编码。
本文所用的术语“衍生物”旨在表示相对于通过SEQ ID NO：55 或SEQ ID NO：56鉴定的野生型VAR2CSA或其片段，显示基本上相同或改善的生物活性的VAR2CSA多肽，其中母体肽的一个或多个氨基酸已被化学修饰，例如通过烷基化，PEG化，酰化，酯形成或酰胺形成等。
术语“构建体”旨在表示可以基于编码目标多肽的完整或部分天然存在的核苷酸序列的多核苷酸片段。构建体可任选地含有其它多核苷酸片段。以类似的方式，术语“可由多核苷酸构建体编码的氨基酸”涵盖可以由上述定义的多核苷酸构建体编码的氨基酸，即氨基酸如 Ala，Val，Leu，Ile，Met，Phe，Trp，Pro，Gly，Ser，Thr，Cys， Tyr，Asn，Glu，Lys，Arg，His，Asp和Gln。
本文所用的术语“载体”是指能够在宿主细胞中扩增的任何核酸实体。因此，载体可以是自主复制载体，即一种载体，其作为染色体外实体存在，其复制独立于染色体复制，例如质粒。或者，载体可以是当被引入宿主细胞时被整合到宿主细胞基因组中并与其已被整合到其中的染色体一起复制的载体。载体的选择通常取决于其被引入的宿主细胞。载体包括但不限于质粒载体，噬菌体载体，病毒或粘粒载体。载体通常含有复制起点和至少一个可选择的基因，即编码易于检测的产物或其存在对细胞生长至关重要的基因。
如本文所用，术语“合适的生长培养基”是指含有培养细胞和编码本发明的VAR2CSA多肽的核酸序列的表达所需的营养物和其它成分的培养基。
在本文中，术语“治疗”意在包括预防，治愈和缓解例如癌症中的涉及CSA的表达例如不适当表达的疾病、病症或病况的症状。因此，根据本发明的VAR2CSA多肽、缀合物或衍生物的预防和治疗性施用包括在术语“治疗”中。
本文所用的术语“受试者”是指任何动物，特别是哺乳动物，例如人，并且可以在适当的情况下与术语“患者”互换使用。
本领域已知的术语“序列同一性”是指通过比较序列确定的两个或更多个多肽分子或两个或更多个核酸分子的序列之间的关系。在本领域中，“同一性”还指根据情况如通过两个或更多个核苷酸残基或两个或更多个氨基酸残基的串之间的匹配数确定的核酸分子之间或多肽之间的序列相关性程度。“同一性”测量具有由特定数学模型或计算机程序(即“算法”)寻址的间隙对齐(如果有的话)的两个或更多个序列中的较小者之间的相同匹配的百分比。
术语“相似度”是相关概念，但与“同一性”不同，是指包括相同匹配和保守替代匹配的序列关系。如果两个多肽序列具有例如(分数(10/20))相同的氨基酸，并且其余部分都是非保守取代，则同一性和相似度百分比均为50％。如果在同一个例子中还有5个保守取代的位置，那么同一性百分比仍然是50％，但相似度百分比为75％((分数(15/20)))。因此，在存在保守取代的情况下，两个多肽之间的相似度将高于这两个多肽之间的同一性百分比。
对SEQ ID NO：1-56、60-70和72-75、128的氨基酸序列的保守修饰(以及对编码核苷酸的相应修饰)将产生具有与天然存在的 VAR2CSA多肽相似的功能和化学特征的VAR2CSA多肽。相比之下， VAR2CSA多肽的功能和/或化学特征的实质性修饰可以通过选择SEQ ID NO：1-56、60-70、72-75和128的氨基酸序列中的取代来完成，这样的取代在其维持(a)取代区域中分子骨架的结构，例如作为片状或螺旋构象，(b)在靶位点处分子的电荷或疏水性，或(c)大部分侧链的作用上显著不同。
例如，“保守氨基酸取代”可以包括用非天然残基取代天然氨基酸残基，使得对该位置上的氨基酸残基的极性或电荷几乎没有影响或没有影响。此外，多肽中的任何天然残基也可以被丙氨酸取代，如先前对于“丙氨酸扫描诱变”所描述的(参见例如MacLennan等人,1998, Acta Physiol.Scand.Suppl.643:55-67；Sasaki等人,1998,Adv. Biophys.35:1-24，其讨论了丙氨酸扫描诱变)。
期望的氨基酸取代(无论是保守的还是非保守的)可以由本领域技术人员在需要这样的取代时确定。例如，氨基酸取代可用于鉴定优选含有VAR2CSA的本发明多肽的重要残基，或增加或降低本文所述的多肽的亲和力。
天然存在的残基可以根据共同的侧链性质分为几类：
1)疏水性：正亮氨酸，Met，Ala，Val，Leu，Ile；
2)中性亲水性：Cys，Ser，Thr，Asn，Gln；
3)酸性：Asp，Glu；
4)碱性：His，Lys，Arg；
5)影响链取向的残基：Gly，Pro；和
6)芳香族：Trp，Tyr，Phe。
例如，非保守替代可能涉及到这些类之一的成员与来自另一类的成员的交换。这种取代的残基可以被引入与非恶性疟原虫VAR2CSA 多肽同源的恶性疟原虫VAR2CSA多肽的区域，或者被引入分子的非同源区域。
在进行这种改变时，可以考虑氨基酸的亲水指数。基于它们的疏水性和电荷特性，每个氨基酸被指定亲水指数，它们是：异亮氨酸 (+4.5)；缬氨酸(+4.2)；亮氨酸(+3.8)；苯丙氨酸(+2.8)；半胱氨酸/胱氨酸(+2.5)；甲硫氨酸(+1.9)；丙氨酸(+1.8)；甘氨酸(-0.4)；苏氨酸(-0.7)；丝氨酸(-0.8)；色氨酸(-0.9)；酪氨酸(-1.3)；脯氨酸(-1.6)；组氨酸(-3.2)；谷氨酸(-3.5)；谷氨酰胺(-3.5)；天冬氨酸(-3.5)；天冬酰胺(-3.5)；赖氨酸(-3.9)；和精氨酸(-4.5)。
亲水性氨基酸指数在赋予蛋白质交互式生物学功能中的重要性在本领域中是得到认可的。Kyte等人,J.Mol.Biol.,157:105-131(1982)。已知某些氨基酸可以替代具有相似亲水指数或分数的其它氨基酸，并且仍然保留类似的生物学活性。在根据亲水指数进行变化时，优选亲水指数在±2以内的氨基酸取代，特别优选在±1以内的氨基酸取代，并且更优选在±0.5以内。
还应理解在本领域中，可以基于亲水性有效地制备类似氨基酸的取代，特别是当由此产生的生物功能等同的蛋白质或肽旨在用于免疫学实施方案中时，如在本文的实例中。由其相邻氨基酸的亲水性决定的蛋白质的最大局部平均亲水性与其免疫原性和抗原性(即与蛋白质的生物学性质)相关。
以下亲水性值被分配给氨基酸残基：精氨酸(+3.0)；赖氨酸('3.0)；天冬氨酸(+3.0±1)；谷氨酸(+3.0±1)；丝氨酸(+0.3)；天冬酰胺 (+0.2)；谷氨酰胺(+0.2)；甘氨酸(0)；苏氨酸(-0.4)；脯氨酸(-0.5±1)；丙氨酸(-0.5)；组氨酸(-0.5)；半胱氨酸(-1.0)；甲硫氨酸(-1.3)；缬氨酸(-1.5)；亮氨酸(-1.8)；异亮氨酸(-1.8)；酪氨酸(-2.3)；苯丙氨酸(-2.5)；色氨酸(-3.4)。在基于相似的亲水性值进行改变时，优选亲水性值在±2以内的氨基酸取代，特别优选在±1以内的氨基酸取代，并且更特别优选在±0.5以内。还可以基于亲水性从主要氨基酸序列鉴定表位。这些区域也称为“表位核心区域”。
技术人员将能够使用公知的技术来确定如SEQ ID NO：1-75和 128所示的多肽的合适变体。为了鉴定可以在不破坏活性的情况下改变的分子的合适区域，本领域技术人员可以针对不被认为对活性重要的区域。例如，当来自相同物种或来自其它物种的具有相似活性的相似多肽是已知的时，本领域技术人员可将VAR2CSA多肽的氨基酸序列与此类相似多肽进行比较。通过这样的比较，可以鉴定在相似多肽中保守的分子的残基和部分。应当理解，相对于这种类似多肽不保守的VAR2CSA多肽区域中的变化将不太可能不利地影响VAR2CSA多肽的生物学活性和/或结构。本领域技术人员还将知道，即使在相对保守的区域中，也可以在保留活性的情况下用化学上相似的氨基酸替代天然存在的残基(保守氨基酸残基取代)。因此，即使对生物活性或结构可能重要的区域也可以进行保守的氨基酸取代而不破坏生物活性或不影响多肽结构。
此外，本领域技术人员可以审查鉴定相似多肽中对于活性或结构重要的残基的结构功能研究。鉴于这样的比较，可以预测VAR2CSA 多肽中对应于对相似多肽的活性或结构重要的氨基酸残基的氨基酸残基的重要性。本领域技术人员可以选择本发明的VAR2CSA多肽和其它多肽的这样预测的重要氨基酸残基的化学上相似的氨基酸取代。
本领域技术人员还可以分析与相似多肽中的该结构相关的三维结构和氨基酸序列。鉴于该信息，本领域技术人员可以相对于其三维结构预测多肽的氨基酸残基的比对。本领域技术人员可以选择不对预测在蛋白质表面上的氨基酸残基进行彻底改变，因为这样的残基可能参与与其它分子的重要相互作用。此外，本领域技术人员可以产生在每个所需氨基酸残基处含有单个氨基酸取代的测试变体。然后可以使用如本文所述的活性测定法筛选变体。这样的变体可用于收集关于合适变体的信息。例如，如果发现特定氨基酸残基的改变导致破坏的，不希望地降低的或不合适的活性，则将避免具有这种变化的变体。换句话说，基于从这种常规实验收集的信息，本领域技术人员可以容易地确定氨基酸，其中应该单独地或与其它突变组合避免进一步的取代。
一些科学出版物专门用于预测二级结构。参见Moult J.,Curr.Op. in Biotech.,7(4):422-427(1996),Chou等人,Biochemistry, 13(2):222-245(1974)；Chou等人,Biochemistry,113(2):211-222(1974)； Chou等人,Adv.Enzymol.Relat.Areas Mol.Biol,47:45-148(1978)； Chou等人,Ann.Rev.Biochem.,47:251-276和Chou等人,Biophys.J.,26:367-384(1979)。此外，计算机程序目前可用于辅助预测二级结构。预测二级结构的一种方法是基于同源建模。例如，具有大于30％的序列同一性或大于40％的相似性的两个多肽或蛋白质通常具有相似的结构拓扑结构。蛋白质结构数据库(PDB)的最近扩展提供了二级结构的增强的可预测性，包括多肽或蛋白质结构内潜在的折叠数量。参见Holm等人,Nucl.Acid.Res.,27(1):244-247(1999)。已经提出 (Brenner等人,Curr.Op.Struct.Biol.,7(3):369-376(1997))，在给定的多肽或蛋白质中存在有限数量的折叠，并且一旦已解析关键的结构数量，结构预测将在准确性上获得巨大的增长。
预测二级结构的其它方法包括“线面法”(Jones,D.,Curr.Opin. Struct.Biol.,7(3):377-87(1997)；Sippl等人,Structure,4(1):15-9 (1996))，“侧貌分析法”(Bowie等人,Science,253:164-170(1991)； Gribskov等人,Meth.Enzymol.,183:146-159(1990)；Gribskov等人, Proc.Nat.Acad.Sci.,84(13):4355-4358(1987))和“进化连接法”(参见Home，同上，和Brenner，同上)。
相关多肽的鉴定和相似性可以通过已知方法容易地计算。这些方法包括但不限于Computational Molecular Biology,Lesk,A.M.,ed., Oxford University Press,NewYork,1988；Biocomputing:Informatics and Genome Projects,Smith,D.W.,ed.,Academic Press,New York, 1993；Computer Analysis of Sequence Data,Part 1,Griffin,A.M., and Griffin,H.G.,eds.,Humana Press,New Jersey,1994；SequenceAnalysis in Molecular Biology,von Heinje,G.,Academic Press,1987； SequenceAnalysis Primer,Gribskov,M.and Devereux,J.,eds.,M. Stockton Press,New York,1991；和Carillo等人,SIAM J.Applied Math.,48:1073(1988)中描述的那些。
确定同一性和/或相似性的优选方法被设计为在测试的序列之间给出最大的匹配。在可公开获得的计算机程序中描述了确定同一性和相似性的方法。用于确定两个序列之间的同一性和相似性的优选计算机程序方法包括但不限于GCG程序包，包括GAP(Devereux等人, Nucl.Acid.Res.,12:387(1984)；Genetics Computer Group,Universityof Wisconsin,Madison,Wis.),BLASTP,BLASTN,和FASTA (Altschul等人,J.Mol.Biol.,215:403-410(1990))。BLASTX程序可从国家生物技术信息中心(NCBI)和其它来源(BLASTManual，Altschul 等人NCB/NLM/NIH Bethesda,Md.20894；Altschul等人，同上)公开获得。公知的Smith Waterman算法也可以用来确定同一性。
用于比对两个氨基酸序列的某些比对方案可以导致两个序列的仅短区域的匹配，并且即使两个全长序列之间没有显著关系，该小的比对区域也可能具有非常高的序列同一性。因此，在优选的实施方案中，所选择的比对方法(GAP程序)将导致跨越靶多肽的至少50个连续氨基酸的比对。
例如，通过使用计算机算法GAP(Genetics Computer Group, University ofWisconsin,Madison,Wis.)，待确定百分比序列同一性的两个多肽被比对以最佳匹配它们各自的氨基酸(“匹配跨度”，由算法确定)。间隙开口罚分(计算为平均对角线的3倍；“平均对角线”是所使用的比较矩阵的对角线的平均值；“对角线”是通过特定比较矩阵分配给每个完美氨基酸匹配的得分或数字)和间隙延伸罚分(通常是间隙开口罚分的{分数(1/10)}倍)以及诸如PAM 250或BLOSUM 62的比较矩阵与算法结合使用。标准比较矩阵(对于PAM250比较矩阵参见Dayhoff等人,Atlas of Protein Sequence and Structure,vol.5,supp.3(1978)；对于BLOSUM 62比较矩阵Henikoff等人,Proc.Natl. Acad.Sci USA,89:10915-10919(1992))也通过算法使用。
多肽序列比较的优选参数包括：
算法：Needleman等人,J.Mol.Biol,48:443-453(1970)；比较矩阵：来自Henikoff等人,Proc.Natl.Acad.Sci.USA,89:10915-10919 (1992)的BLOSUM 62；间隙罚分：12，间隙长度罚分：4，相似度阈值：0。
GAP程序对上述参数是可用的。上述参数是使用GAP算法的多肽比较的默认参数(以及对于末端间隙无罚分)。
核酸分子序列比较的优选参数包括：算法：Needleman等人,J. Mol Biol.,48:443-453(1970)；比较矩阵：匹配＝+10，错配＝0，间隙罚分：50，间隙长度罚分：3。
GAP程序对上述参数也是可用的。上述参数是核酸分子比较的默认参数。
可以使用其它示例性算法，间隙开口罚分，间隙延伸罚分，比较矩阵，相似性阈值等，包括在Program Manual,Wisconsin Package, Version 9,September,1997中列出的那些。要做出的具体选择对于本领域技术人员将是显而易见的，并且将取决于进行的具体比较，例如 DNA与DNA，蛋白质与蛋白质，蛋白质与DNA的对比；此外，比较是否在给定的序列对之间(在这种情况下通常优选为GAP或BestFit) 或在序列的一个序列和大型数据库之间(在这种情况下优选为FASTA 或BLASTA)。
本发明的发明人已经解决并找到了与PM中胎盘粘附的分子机理相关的以下关键问题的答案：1)描述了与VAR2CSA序列相关的差异CSA粘附，2)什么是VAR2CSA与CSA结合的精确的最小结构要求，3)VAR2CSA与CSA之间存在什么类型的化学相互作用，和最后4)该信息是否可用于设计最佳疫苗抗原？
通过表达相同的FCR3和3d7 VAR2CSA截短提，本发明人显示 VAR2CSA以相似的亲和力和特异性结合CSA，而不管寄生虫株系来源。这两个序列具有79.6％的序列同一性。本发明人进一步证明，高 CSA结合亲和力保留在几个较短的片段中，并且包含来自侧翼域间的小区域的DBL2X形成包含高亲和性CSA结合位点的紧密核心。本发明人在计算机中定义了VAR2CSA中的推定的GAG结合位点，并且通过缺失和取代，本发明人发现这些位点的突变对CSPG结合没有影响。通过使用聚电解质-蛋白质相互作用的理论，本发明人已经表明，VAR2CSA-CSA相互作用可能不仅仅取决于离子相互作用。最后，本发明人已经显示出几种短VAR2CSA片段能够诱导粘附-阻断抗体的产生，并且抗粘附抗体靶向所提出的CSA结合区域。这些数据提供了对PfEMP1分子与其配体之间的相互作用的生物化学性质的第一次详细了解。
制备本发明的多肽
本文描述的本发明的VAR2CSA多肽和其它多肽可以通过重组核酸技术并如WO2013/117705中所述来产生。通常，修饰克隆的野生型 VAR2CSA核酸序列以编码所需的蛋白质。然后将该修饰的序列插入到表达载体中，其进而被转化或转染到宿主细胞中。高等真核细胞，特别是培养的哺乳动物细胞可用作宿主细胞。也可以使用原核细胞例如乳酸乳球菌或大肠杆菌来表达多肽，只要这些原核生物能够产生二硫键或蛋白质被正确地重新折叠。此外，酵母菌株也可以用于表达蛋白质，包括酿酒酵母和毕赤酵母。
氨基酸序列改变可以通过多种技术来完成。核酸序列的修饰可以是通过位点特异性诱变。用于位点特异性诱变的技术是本领域熟知的，并且描述于例如Zoller and Smith(DNA 3:479-488,1984)或"Splicing by extension overlap",Horton等人,Gene 77,1989,pp.61-68中。因此，通过使用VAR2CSA的核苷酸和氨基酸序列，可以引入选择的改变。同样，利用特异性引物使用聚合酶链反应制备DNA构建体的方法是本领域技术人员熟知的(参见PCR Protocols,1990,Academic Press,San Diego,California,USA。
本发明的多肽还可以包含非天然存在的氨基酸残基。非天然存在的氨基酸包括但不限于β-丙氨酸，去氨基组氨酸，反式-3-甲基脯氨酸， 2,4-甲脯氨酸，顺式-4-羟脯氨酸，反式-4-羟脯氨酸，N-甲基甘氨酸，别苏氨酸，甲基苏氨酸，羟乙基半胱氨酸，羟乙基高半胱氨酸，硝基谷氨酰胺，高谷氨酰胺，哌啶酸，噻唑烷羧酸，脱氢脯氨酸，3-和4- 甲基脯氨酸，3,3-二甲基脯氨酸，叔亮氨酸，正缬氨酸，2-氮杂苯丙氨酸，3-氮杂苯丙氨酸，4-氮杂苯丙氨酸和4-氟苯丙氨酸。本领域已知几种方法将非天然存在的氨基酸残基并入多肽中。例如，可以使用体外系统，其中使用化学氨酰化的抑制性tRNA抑制无义突变。合成氨基酸和氨基酰化tRNA的方法是本领域已知的。含有无义突变的质粒的转录和翻译在包含大肠杆菌S30提取物和市售的酶和其它试剂的无细胞系统中进行。多肽通过色谱纯化。参见，例如，Robertson等人,J. Am.Chem.Soc.113:2722,1991；Ellman等人,Methods Enzymol. 202:301,1991；Chung等人,Science 259:806-9,1993；Chung等人, Proc.Natl.Acad.Sci.USA90:10145-9,1993。在第二种方法中，通过显微注射突变的mRNA和化学氨基酰化的抑制性tRNA在非洲爪蟾中进行翻译(Turcatti等人,J.Biol.Chem.271:19991-8,1996)。在第三种方法中，大肠杆菌细胞在不存在需要被置换的天然氨基酸(例如苯丙氨酸)的情况下，并且在存在所需的非天然存在的氨基酸(例如2- 氮杂苯丙氨酸，3-氮杂苯丙氨酸，4-氮杂苯丙氨酸或4-氟苯丙氨酸) 的情况下培养。将非天然存在的氨基酸并入多肽中代替其天然对应物。参见Koide等人,Biochem.33:7470-6,1994。天然存在的氨基酸残基可以通过体外化学修饰转化为非天然存在的物质。化学修饰可以与定点诱变组合以进一步扩大取代范围(Wynn and Richards,Protein Sci. 2:395-403,1993)。
编码本发明的本发明的VAR2CSA多肽和其它多肽的核酸构建体可以适当地为基因组或cDNA来源，例如通过制备基因组或cDNA文库并通过使用合成寡核苷酸探针根据标准技术杂交来筛选编码全部或部分多肽的DNA序列(参见Sambrook等人,Molecular Cloning:ALaboratory Manual,2nd.Ed.Cold Spring Harbor Labora-tory,Cold Spring Harbor,New York,1989)。
编码VAR2CSA多肽的核酸构建体也可以通过建立的标准方法合成制备，例如由Beaucage and Caruthers,Tetrahedron Letters 22 (1981),1859-1869描述的亚磷酰胺方法，由Matthes等人,EMBO Journal 3(1984),801-805描述的方法。根据亚磷酰胺方法，寡核苷酸例如在自动DNA合成仪中合成，纯化，退火，连接并克隆在合适的载体中。编码恶性疟原虫VAR2CSA多肽和本发明的其它多肽的 DNA序列也可以通过使用特异性引物的聚合酶链反应来制备，例如 US 4,683,202，Saiki等人，Science 239(1988)，487-491，或Sambrook等人，同上。
此外，核酸构建体可以是混合的合成和基因组、混合的合成和 cDNA或混合的基因组和cDNA来源，其按照标准技术通过连接合成、基因组或cDNA来源的片段(视情况而定)、对应于整个核酸构建体各个部分的片段来制备。
核酸构建体优选为DNA构建体。用于生产根据本发明的 VAR2CSA多肽和其它多肽的DNA序列通常将在VAR2CSA的氨基末端编码前原多肽，以获得适当的翻译后加工和从宿主细胞分泌。
根据本发明的编码恶性疟原虫VAR2CSA多肽和其它多肽的 DNA序列通常插入可以是任何载体的重组载体，其可以方便地经历重组DNA程序，并且载体的选择通常将取决于要引入的宿主细胞。因此，载体可以是自主复制载体，即一种载体，其作为染色体外实体存在，其复制独立于染色体复制，例如质粒。或者，载体可以是当被引入宿主细胞时被整合到宿主细胞基因组中并与其已被整合到其中的染色体一起复制的载体。
载体优选是表达载体，其中编码恶性疟原虫VAR2CSA多肽和根据本发明的其它多肽的DNA序列可操作地连接到DNA转录所需的其它片段。通常，表达载体衍生自质粒或病毒DNA，或可以含有这两者的元件。术语“可操作地连接”表示这些片段被布置成使得它们配合作用于其预期目的，例如，转录起始于启动子，并通过编码多肽的DNA 序列向前进行。
用于表达根据本发明的VAR2CSA多肽和其它多肽的表达载体将包含能够引导克隆的基因或cDNA的转录的启动子。启动子可以是任何DNA序列，其显示在所选宿主细胞中的转录活性，并且可以衍生自编码与宿主细胞同源或异源的蛋白质的基因。
在哺乳动物细胞中用于引导编码恶性疟原虫VAR2CSA多肽的DNA的转录的合适的启动子的实例是SV40启动子(Subramani等人, Mol.Cell Biol.1(1981),854-864)，MT-1(金属硫蛋白基因)启动子(Palmiter等人,Science 222(1983),809-814)，CMV启动子(Boshart 等人,Cell 41:521-530,1985)或腺病毒2主要晚期启动子(Kaufman and Sharp,Mol.Cell.Biol,2:1304-1319,1982)。
用于昆虫细胞的合适启动子的实例是多角体蛋白启动子(US 4,745,051；Vasuvedan等人,FEBS Lett.311,(1992)7-11)，P10启动子(J.M.Vlak等人,J.Gen.Virology69,1988,pp.765-776)，苜蓿银纹夜蛾核型多角体病毒碱性蛋白启动子(EP 397 485)，杆状病毒立即早期基因1启动子(US 5,155,037；US 5,162,222)或杆状病毒39K 延迟早期基因启动子(US 5,155,037；US 5,162,222)。
适用于酵母宿主细胞的启动子的实例包括来自酵母糖酵解基因的启动子(Hitzeman等人,J.Biol.Chem.255(1980),12073-12080； Alber and Kawasaki,J.Mol.Appl.Gen.1(1982),419-434)或醇脱氢酶基因(Young等人,in GeneticEngineering of Microorganisms for Chemicals(Hollaender等人,eds.),PlenumPress,New York,1982)或 TPI1(US 4,599,311)或ADH2-4c(Russell等人,Nature 304(1983),652 -654)启动子。
用于丝状真菌宿主细胞的合适的启动子的实例是例如ADH3启动子(McKnight等人,The EMBO J.4(1985),2093-2099)或tpiA启动子。其它有用的启动子的实例是衍生自编码米曲霉(A.oryzae)TAKA 淀粉酶，米赫根毛霉(Rhizomucor miehei)天冬氨酸蛋白酶，黑曲霉 (A.niger)中性α-淀粉酶，黑曲霉酸稳定性α-淀粉酶，黑曲霉或泡盛曲霉(A.awamori)葡糖淀粉酶(gluA)，米赫根毛霉脂肪酶，米曲霉碱性蛋白酶，米曲霉丙糖磷酸异构酶或构巢曲霉乙酰胺酶的基因的那些。优选的是TAKA-淀粉酶和gluA启动子。合适的启动子在例如EP238 023和EP 383 779中被提及。
根据需要，编码恶性疟原虫VAR2CSA多肽和其它多肽的DNA 序列也可以可操作地连接到合适的终止子，例如人生长激素终止子(Palmiter等人,Science 222,1983,pp.809-814)或TPI1(Alber and Kawasaki,J.Mol.Appl.Gen.1,1982,pp.419-434)或ADH3(McKnight等人,The EMBO J.4,1985,pp.2093-2099)终止子。表达载体还可以含有位于启动子下游和VAR2CSA序列本身的插入位点的上游的一组RNA剪接位点。可以从腺病毒和/或免疫球蛋白基因获得优选的RNA剪接位点。表达载体中还包含位于插入位点下游的聚腺苷酸化信号。特别优选的聚腺苷酸化信号包括来自SV40(Kaufman 和Sharp，同上)的早期或晚期多腺苷酸化信号，来自腺病毒5Eb区域的多腺苷酸化信号，人生长激素基因终止子(DeNoto等人Nucl. Acids Res.9:3719-3730,1981)或来自恶性疟原虫、人或牛基因的多腺苷酸化信号。表达载体还可以包括位于启动子和RNA剪接位点之间的非编码病毒前导序列，例如腺病毒2三联前导序列；和增强子序列，例如SV40增强子。
为了将本发明的恶性疟原虫VAR2CSA多肽和其它多肽引导到宿主细胞的分泌途径中，可以在重组载体中提供分泌信号序列(也称为前导序列，前原序列或前序列)。分泌信号序列在正确的阅读框架中连接至编码根据本发明的恶性疟原虫VAR2CSA多肽和其它多肽的DNA序列。分泌信号序列通常位于编码肽的DNA序列5'。分泌信号序列可以通常与蛋白质相关联或可以来自编码另一种分泌蛋白质的基因。
为了从酵母细胞分泌，分泌信号序列可以编码任何信号肽，其确保表达的根据本发明的恶性疟原虫VAR2CSA多肽和其它多肽的有效导向进入细胞的分泌途径。信号肽可以是天然存在的信号肽或其功能部分，或其也可以是合成肽。已经发现合适的信号肽是α因子信号肽 (参见US 4,870,008)，小鼠唾液淀粉酶的信号肽(参见O. Hagenbuchle等人,Nature 289,1981,pp.643-646)，修饰的羧肽酶信号肽(参见L.A.Valls等人,Cell 48,1987,pp.887-897)，酵母BAR1 信号肽(参见WO 87/02670)或酵母天冬氨酸蛋白酶3(YAP3)信号肽(参见M.Egel-Mitani等人,Yeast 6,1990,pp.127-137)。
为了在酵母中有效分泌，编码前导肽的序列也可以插入信号序列的下游和编码本发明的恶性疟原虫VAR2CSA多肽和其它多肽的 DNA序列的上游。前导肽的功能是允许表达的肽从内质网引导到高尔基体，并进一步分泌到分泌囊泡中，以分泌到培养基中(即根据本发明的恶性疟原虫VAR2CSA多肽和其它多肽跨细胞壁或至少通过细胞膜进入酵母细胞的周质空间)。前导肽可以是酵母α-因子前导序列(其用途描述于例如US 4,546,082，US 4,870,008，EP 16 201，EP 123 294， EP 123 544和EP 163 529中)。或者，前导肽可以是合成的前导肽，其是在自然界中未发现的前导肽。合成前导肽可以例如如WO 89/02463或WO 92/11378中所述构建。
为了在丝状真菌中使用，信号肽可以方便地来自编码曲霉属淀粉酶或葡糖淀粉酶的基因，编码米赫根毛霉脂肪酶或蛋白酶或柔毛腐质霉(Humicola lanuginosa)脂肪酶的基因。信号肽优选衍生自编码米曲霉TAKA淀粉酶、黑曲霉中性α-淀粉酶、黑曲霉酸稳定淀粉酶或黑曲霉葡糖淀粉酶的基因。合适的信号肽公开于，例如，EP 238 023 和EP 215 594。
为了在昆虫细胞中使用，信号肽可以方便地衍生自昆虫基因(参见WO 90/05783)，例如鳞翅目烟草天蛾脂肪动力激素前体信号肽(参见US 5,023,328)。
用于分别连接编码根据本发明的恶性疟原虫VAR2CSA多肽和其它多肽的DNA序列、启动子和任选地终止子和/或分泌信号序列并将其插入含有复制所需信息的合适载体中的程序是本领域技术人员公知的(参见例如Sambrook等人,Molecular Cloning:ALaboratory Manual,Cold Spring Harbor,New York,1989)。
转染哺乳动物细胞和表达引入细胞的DNA序列的方法描述于例如Kaufman andSharp,J.Mol.Biol.159(1982),601-621；Southern and Berg,J.Mol.Appl.Genet.1(1982),327-341；Loyter等人,Proc. Natl.Acad.Sci.USA 79(1982),422-426；Wigler等人,Cell 14(1978), 725；Corsaro and Pearson,Somatic Cell Genetics 7(1981),603,Graham and van der Eb,Virology 52(1973),456；和Neumann等人, EMBO J.1(1982),841–845中。
将克隆的DNA序列通过例如磷酸钙介导的转染(Wigler等人,Cell 14:725-732,1978；Corsaro and Pearson,Somatic Cell Genetics 7:603-616,1981；Graham and Vander Eb,Virology 52d:456-467,1973) 或电穿孔(Neumann等人,EMBO J.1:841-845,1982)引入培养的哺乳动物细胞中。为了鉴定和选择表达外源DNA的细胞，赋予选择性表型(选择性标记)的基因通常与感兴趣的基因或cDNA一起引入细胞。优选的选择性标记包括赋予对药物例如新霉素、潮霉素和甲氨蝶呤的抗性的基因。可选择标记可以是可扩增的选择性标记。优选的可扩增的选择性标记是二氢叶酸还原酶(DHFR)序列。Thilly(Mammalian CellTechnology,Butterworth Publishers,Stoneham,MA, incorporated herein byreference，其通过引用并入本文)综述了选择性标记。本领域技术人员将容易地选择合适的选择性标记。
选择性标记可以与感兴趣的基因在分开的质粒上同时引入细胞，或者它们可以被引入到相同的质粒上。如果在相同的质粒上，选择性标记和感兴趣的基因可能在不同的启动子或相同的启动子的控制下，后一种布置产生双顺反子信息。这种类型的构建体在本领域中是已知的(例如，Levinson和Simonsen，U.S.4,713,339)。将额外的DNA (称为“载体DNA”)添加到引入细胞的混合物中也可能是有利的。
细胞吸收DNA后，将它们生长在合适的生长培养基中，通常1-2 天，以开始表达感兴趣的基因。如本文所用，术语“合适的生长培养基”是指含有细胞生长和感兴趣的恶性疟原虫VAR2CSA多肽的表达所需的营养物质和其它成分的培养基。培养基通常包括碳源，氮源，必需氨基酸，必需的糖，维生素，盐，磷脂，蛋白质和生长因子。然后应用药物选择以针对稳定地表达选择性标记的细胞的生长进行选择。对于已经用可扩增的选择性标记转染的细胞，可以增加药物浓度以针对增加的克隆序列拷贝数进行选择，从而增加表达水平。然后对稳定转染的细胞的克隆针对表达感兴趣的恶性疟原虫VAR2CSA多肽进行筛选。
其中引入编码根据本发明的恶性疟原虫VAR2CSA多肽和其它多肽的DNA序列的宿主细胞可以是能够产生翻译后修饰的多肽的任何细胞并且包括酵母，真菌和更高等的真核细胞。
用于本发明的哺乳动物细胞系的实例是COS-1(ATCC CRL1650)，幼仓鼠肾(BHK)和293(ATCC CRL 1573；Graham 等人,J.Gen.Virol.36:59-72,1977)细胞系。优选的BHK细胞系是 tk-ts13BHK细胞系(Waechter and Baserga,Proc.Natl.Acad.Sci. USA 79:1106-1110,1982，通过引用并入本文)，以下称为BHK 570 细胞。BHK 570细胞系已经保藏在美国典型培养物保藏中心，12301 Parklawn Dr.,Rockville,Md.20852，ATCC保藏号为CRL10314。tk- ts13BHK细胞系也可从ATCC以保藏号CRL 1632获得。此外，本发明中可以使用许多其它细胞系，包括Rat Hep I(大鼠肝癌；ATCC CRL1600)，大鼠Hep II(大鼠肝癌；ATCCCRL1548)，TCMK(ATCC CCL 139)，人肺(ATCC HB 8065)，NCTC1469(ATCC CCL 9.1)， CHO(ATCC CCL 61)和DUKX细胞(Urlaub and Chasin,Proc.Natl. Acad.Sci.USA 77:4216-4220,1980)。
合适的酵母细胞的实例包括酵母属(Saccharomyces spp.)和裂殖酵母属(Schizosac-charomyces spp.)的细胞，特别是酿酒酵母 (Saccharomyces cerevisiae)或克氏酵母(Saccharomyces kluyveri) 的菌株。用异源DNA转化酵母细胞并从中产生异源多肽的方法描述于例如，US 4,599,311，US 4,931,373，US 4,870,008，5,037,743和US 4,845,075中，所有这些都通过引用并入本文。通过由选择性标记确定的表型选择转化的细胞，通常是耐药性或在不存在特定营养物例如亮氨酸的情况下生长的能力。用于酵母的优选载体是US 4,931,373中公开的POT1载体。根据本发明的编码恶性疟原虫VAR2CSA多肽和其它多肽的DNA序列之前可以有信号序列和任选地前导序列，例如如上文所述的。合适的酵母细胞的其它实例是克鲁维酵母属例如乳酸克鲁维酵母(K.lactis)、汉逊酵母属(Hansenula)例如多形汉逊酵母(H.polymorpha)或毕赤酵母属例如巴斯德毕赤酵母(P.pastoris) 的菌株(参见Gleeson等人,J.Gen.Microbiol.132,1986,pp. 3459-3465；US4,882,279)。
其它真菌细胞的实例是丝状真菌的细胞，例如。曲霉属 (Aspergillus spp.)，脉孢菌属(Neurospora spp.)，镰刀菌属(Fusarium spp.)或木霉属(Trichoderma spp.)，特别是米曲霉、构巢曲霉或黑曲霉的菌株。使用曲霉菌属用于蛋白质的表达描述于例如，EP272 277，EP 238 023，EP 184 438中。例如，可以如Malardier等人,1989, Gene 78:147-156所述进行尖孢镰孢(F.oxysporum)的转化。木霉属的转化可以例如如EP 244 234中所述执行。
当丝状真菌用作宿主细胞时，可以用本发明的DNA构建体进行转化，方便地通过将DNA构建体整合到宿主染色体中以获得重组宿主细胞。通常认为这种整合是有利的，因为DNA序列更可能在细胞中得到稳定的维持。可以根据常规方法例如，通过同源或异源重组将DNA构建体整合到宿主染色体中。
昆虫细胞的转化和其中的异源多肽的产生可以如US 4,745,051； US 4,879,236；US 5,155,037；5,162,222；EP 397,485中所述，其全部内容通过引用并入本文。用作宿主的昆虫细胞系可以适当地是鳞翅目细胞系，例如草地夜蛾(Spodoptera frugiperda)细胞或粉纹夜蛾 (Trichoplusia ni)细胞(参见US 5,077,214)。培养条件可以适当地如WO 89/01029或WO 89/01028或任何上述参考文献所述。
然后在允许恶性疟原虫VAR2CSA多肽表达的条件下，在合适的营养培养基中培养上述转化或转染的宿主细胞，之后从培养物中回收所有或部分所得肽。用于培养细胞的培养基可以是适于生长宿主细胞的任何常规培养基，例如含有适当补充物的最小或复杂培养基。合适的培养基可从商业供应商获得，或者可以根据公开的配方制备(例如在美国典型培养物保藏中心的目录中)。然后可以通过常规方法从培养基中回收由细胞产生的恶性疟原虫VAR2CSA多肽，包括通过离心或过滤从培养基中分离宿主细胞，通过盐例如硫酸铵沉淀上清液或滤液的蛋白水性组分，通过各种色谱法纯化，例如离子交换层析，凝胶层析，亲和层析等，这取决于所涉及的多肽的类型。
转基因动物技术可用于产生本发明的VAR2CSA多肽和其它多肽。优选在宿主雌性哺乳动物的乳腺内产生蛋白质。乳腺中的表达以及随后将感兴趣的蛋白分泌到乳中克服了从其它来源分离蛋白质所遇到的许多困难。乳容易收集，可获得的量大，并且生物化学表征良好。此外，主要的乳蛋白以高浓度(通常为约1至15g/l)存在于乳中。
从商业角度来看，显然优选使用具有大产乳量的物种作为宿主。虽然可以使用较小的动物如小鼠和大鼠(并且在原理证明阶段是优选的)，但是优选使用家畜哺乳动物，包括但不限于猪，山羊，绵羊和牛。绵羊是特别优选的，这是因为以下因素：在这种物种中转基因的先前历史，乳产率，成本和用于收集绵羊乳的设备的可用性(对于影响宿主物种的选择的因素的比较参见例如WO 88/00239)。通常期望选择一些已经饲养用于产乳的宿主动物，例如East Friesland羊，或者在后期通过育种转基因品系来引入奶畜。无论如何，应该使用已知健康状况良好的动物。
为了在乳腺中获得表达，使用来自乳蛋白基因的转录启动子。乳蛋白基因包括编码酪蛋白(参见美国专利5,304,489)，β乳球蛋白，乳清蛋白和乳清酸性蛋白的那些基因。β乳球蛋白(BLG)启动子是优选的。在羊β乳球蛋白基因的情况下，通常使用基因的5'侧翼序列的至少近端406bp的区域，尽管5'侧翼序列的大部分最高约5kbp是优选的，例如包含5'侧翼启动子和β乳球蛋白基因的非编码部分的 DNA片段(参见Whitelaw等，Biochem.J.286：31 39(1992))。来自其它物种的启动子DNA的类似片段也是合适的。
β乳球蛋白基因的其它区域也可以掺入构建体中，如待表达基因的基因组区域。在本领域中普遍接受的是，缺乏内含子的构建体例如与含有这些DNA序列的构建体相比表达差(参见Brinster等人,Proc. Natl.Acad.Sci.USA 85:836 840(1988)；Palmiter等人,Proc.Natl. Acad.Sci.USA 88:478 482(1991)；Whitelaw等人,Transgenic Res.1: 3 13(1991)；WO 89/01343；和WO 91/02318，各自其通过引用并入本文)。在这方面，在可能的情况下通常优选使用含有编码感兴趣的蛋白质或多肽的基因的全部或部分天然内含子的基因组序列，因此进一步包含来自例如β乳球蛋白基因的至少一些内含子是优选的。一个这样的区域是提供来自羊β乳球蛋白基因的3'非编码区的内含子剪接和 RNA多腺苷酸化的DNA区段。当代替基因的天然3'非编码序列时，该羊β乳球蛋白片段可以增强和稳定感兴趣的蛋白质或多肽的表达水平。在其它实施方案中，围绕VAR2CSA序列的起始ATG的区域被来自乳特异性蛋白质基因的相应序列替换。这种替代提供了推定的组织特异性起始环境以增强表达。替代整个VAR2CSA前原和5'非编码序列为例如BLG基因的那些序列是方便的，尽管较小的区域可以被替换。
为了在转基因动物中表达本发明的VAR2CSA多肽和其它多肽，编码VAR2CSA的DNA片段与其表达所需的其它DNA区段可操作地连接以产生表达单元。这样的其它区段包括上文所述的启动子，以及提供转录终止和mRNA的多聚腺苷酸化的序列。表达单元还将包括与编码修饰的VAR2CSA的区段可操作地连接的编码分泌信号序列的 DNA区段。分泌信号序列可以是天然分泌信号序列，或者可以是另一种蛋白质，例如乳蛋白质的分泌信号序列(参见例如von Heijne,Nucl. Acids Res.14:4683 4690(1986)；和Meade等人,U.S.4,873,316，其通过引用并入本文)。
用于转基因动物的表达单元的构建通过将VAR2CSA序列插入含有其它DNA区段的质粒或噬菌体载体中进行，尽管表达单元可以通过基本上任何连接序列构建。提供含有编码乳蛋白质的DNA片段的载体并且替换乳蛋白的编码序列为VAR2CSA变体的编码序列是特别方便的；从而产生包含乳蛋白基因的表达调控序列的基因融合体。无论如何，在质粒或其它载体中克隆表达单元有助于VAR2CSA序列的扩增。扩增方便地在细菌(例如大肠杆菌)宿主细胞中进行，因此载体通常包括复制起点和在细菌宿主细胞中有功能的选择标记。然后将表达单元引入所选宿主物种的受精卵(包括早期胚胎)中。引入异源 DNA可以通过几种途径之一完成，包括显微注射(例如美国专利号 4,873,191)，逆转录病毒感染(Jaenisch,Science 240:1468 1474(1988)) 或使用胚胎干(ES)细胞的位点定向整合(由Bradley等人, Bio/Technology 10:534 539(1992)综述)。然后将卵子植入假妊娠雌性的输卵管或子宫，并允许发育成型。在其生殖系中携带引入的DNA 的后代可以以正常的孟德尔方式将DNA转移到其后代，从而允许转基因群的发育。生产转基因动物的一般方法是本领域已知的(参见例如Hogan等人,Manipulating the Mouse Embryo:A Laboratory Manual,Cold SpringHarbor Laboratory,1986；Simons等人, Bio/Technology 6:179 183(1988)；Wall等人,Biol.Reprod.32:645 651(1985)；Buhler等人,Bio/Technology 8:140 143(1990)；Ebert等人, Bio/Technology 9:835 838(1991)；Krimpenfort等人,Bio/Technology 9:844 847(1991)；Wall等人,J.Cell.Biochem.49:113 120(1992)；U.S. 4,873,191；U.S.4,873,316；WO 88/00239,WO 90/05188,WO 92/11757；和GB 87/00458)。将外源DNA序列引入哺乳动物及其生殖细胞的技术最初在小鼠中开发(参见例如，Gordon等人,Proc.Natl.Acad.Sci.USA 77:7380 7384(1980)；Gordon and Ruddle,Science 214:1244 1246(1981)；Palmiterand Brinster,Cell 41:343 345(1985)；Brinster 等人,Proc.Natl.Acad.Sci.USA 82:4438 4442(1985)；和Hogan等(同上))。这些技术随后适用于大型动物，包括家畜物种(参见例如WO 88/00239，WO 90/05188和WO 92/11757；和Simons等人, Bio/Technology 6:179183(1988))。总而言之，在目前用于生产转基因小鼠或家畜的最有效的途径中，根据已建立的技术将感兴趣的DNA 的数百个线性分子注射入受精卵的原核之一。也可以使用将DNA注射到受精卵的细胞质中。
也可以使用转基因植物的生产。表达可以被泛化或定向到特定的器官，例如块茎(参见，Hiatt,Nature 344:469 479(1990)；Edelbaum 等人,J.Interferon Res.12:449453(1992)；Sijmons等人, Bio/Technology 8:217 221(1990)；和EP 0 255 378)。
施用和药物组合物
组合治疗
本说明书中定义的优选包含VAR2CSA的多肽、衍生物或缀合物可以与一种或多种其它癌症试剂同时或相继施用，和/或与其它已知疗法组合使用。这些因素可以以单剂型提供，其中单剂型包含两种化合物，或以包含VAR2CSA多肽的制剂作为第一单位剂型以及一种或更多其它化合物的制剂作为第二单位剂型的组分试剂盒的形式提供。当在本说明书中提及第一或第二或第三等单位剂型时，这并不表示优选的施用顺序，而是为了方便起见而进行。
可以与VAR2CSA多肽组合使用的合适的其它癌症药物或疗法包括已经在市场上或在开发中的抗体，包括维莫拉本(Hoffmann-La Roche)，针对MCSP的人单克隆抗体，Therapeutical(Micromet Inc) 抗MCSP，其使用BiTE抗体平台技术，以及对MCSP具有特异性的细胞毒性T细胞的继代转移。
“同时”给药VAR2CSA多肽的制剂和一种或多种其它化合物的制剂是指以单剂型施用化合物，或施用第一药剂，然后施用第二药剂，其时间间隔不超过15分钟，优选10分钟，更优选5分钟，更优选2 分钟。任何一个因素都可以先施用。
“相继”给药是指施用第一药剂，然后施用第二药剂，其时间间隔超过15分钟。可以首先施用两个单位剂型中的任一种。优选地，通过相同的静脉内通路注射两种产品。
本发明的另一个目的是提供包含根据本发明的多肽的药物制剂，优选含有以0mg/ml至1mg/ml的血清/血浆浓度存在的VAR2CSA，并且其中制剂具有2.0到10.0的pH。制剂还可以包含缓冲体系，防腐剂，张度剂，螯合剂，稳定剂和表面活性剂。在本发明的一些实施方案中，药物制剂是水性制剂，即包含水的制剂。这种制剂通常是溶液或悬浮液。在本发明的另一个实施方案中，药物制剂是水溶液。术语“水性制剂”定义为包含至少50％w/w水的制剂。同样，术语“水溶液”定义为包含至少50％w/w水的溶液，术语“水性悬浮液”定义为包含至少50％w/w水的悬浮液。
在其它实施方案中，药物制剂是冷冻干燥的制剂，医师或患者在使用前向其加入溶剂和/或稀释剂。
在其它实施方案中，药物制剂是可在没有任何先前溶解的情况下使用的即用干燥制剂(例如冷冻干燥或喷雾干燥的)。
在另一方面，本发明涉及包含含有VAR2CSA的多肽的水溶液和缓冲液的药物制剂，其中含有VAR2CSA的多肽以0-1mg/ml或更高的血清/血浆浓度存在，其中制剂具有约2.0至约10.0的pH。
在本发明的其它实施方案中，制剂的pH选自2.0,2.1,2.2,2.3,2.4, 2.5,2.6,2.7,2.8,2.9,3.0,3.1,3.2,3.3,3.4,3.5,3.6,3.7,3.8,3.9,4.0,4.1, 4.2,4.3,4.4,4.5,4.6,4.7,4.8,4.9,5.0,5.1,5.2,5.3,5.4,5.5,5.6,5.7,5.8, 5.9,6.0,6.1,6.2,6.3,6.4,6.5,6.6,6.7,6.8,6.9,7.0,7.1,7.2,7.3,7.4,7.5, 7.6,7.7,7.8,7.9,8.0,8.1,8.2,8.3,8.4,8.5,8.6,8.7,8.8,8.9,9.0,9.1,9.2, 9.3,9.4,9.5,9.6,9.7,9.8,9.9和10.0。
在本发明的另一个实施方案中，缓冲液选自乙酸钠，碳酸钠，柠檬酸盐，甘氨酰甘氨酸，组氨酸，甘氨酸，赖氨酸，精氨酸，磷酸二氢钠，磷酸氢二钠，磷酸钠和三(羟甲基)氨基甲烷，二甘氨酸，三甲基甘氨酸，苹果酸，琥珀酸，马来酸，富马酸，酒石酸，天冬氨酸或其混合物。这些特定缓冲液中的每一个构成本发明的替代实施方案。
在本发明的另一个实施方案中，制剂还包含药学上可接受的防腐剂。在本发明的另一个实施方案中，防腐剂选自苯酚，邻甲酚，间甲酚，对甲酚，对羟基苯甲酸甲酯，对羟基苯甲酸丙酯，2-苯氧基乙醇，对羟基苯甲酸丁酯，2-苯乙醇，苯甲醇，氯丁醇和硫代吗啉，布洛芬醇，苯甲酸，亚胺脲，氯己定，脱氢乙酸钠，氯甲酚，对羟基苯甲酸乙酯，苄索氯铵，氯苯啶(3p-氯苯氧基丙烷-1,2-二醇)或其混合物。在本发明的另一个实施方案中，防腐剂以0.1mg/ml至20mg/ml的浓度存在。在本发明的另一个实施方案中，防腐剂以0.1mg/ml至5mg/ml 的浓度存在。在本发明的另一个实施方案中，防腐剂以5mg/ml至10mg/ml的浓度存在。在本发明的另一个实施方案中，防腐剂以 10mg/ml至20mg/ml的浓度存在。这些特定防腐剂中的每一种构成本发明的替代实施方案。在药物组合物中使用防腐剂是本领域技术人员所熟知的。为方便起见，参考Remington:The Science and Practice of Pharmacy,19thedition,1995。
在本发明的另一个实施方案中，制剂还包含等渗剂。在本发明的另一个实施方案中，等渗剂选自盐(例如氯化钠)，糖或糖醇，氨基酸(例如L-甘氨酸，L-组氨酸，精氨酸，赖氨酸，异亮氨酸，天冬氨酸，色氨酸，苏氨酸)，糖醇(例如甘油(丙三醇)，1,2-丙二醇(丙二醇)，1,3-丙二醇，1,3-丁二醇)聚乙二醇(例如PEG400)或其混合物。可以使用任何糖如单糖，二糖或多糖或水溶性葡聚糖，包括例如果糖，葡萄糖，甘露糖，山梨糖，木糖，麦芽糖，乳糖，蔗糖，海藻糖，葡聚糖，支链淀粉，糊精，环糊精，可溶性淀粉，羟乙基淀粉和羧甲基纤维素钠。在一些实施方案中，糖添加剂是蔗糖。糖醇被定义为具有至少一个-OH基团的C4-C8烃，并且包括例如甘露糖醇，山梨糖醇，肌醇，半乳糖醇，二糖醇，木糖醇和阿拉伯糖醇。在一些实施方案中，糖醇添加剂是甘露糖醇。上述糖或糖醇可以单独使用或组合使用。只要糖或糖醇在液体制剂中溶解，并且不会对使用本发明的方法达到的稳定效果产生不利影响，则所用量没有固定的限制。在一些实施方案中，糖或糖醇浓度为约1mg/ml至约150mg/ml。在本发明的另一个实施方案中，等渗剂以1mg/ml至50mg/ml的浓度存在。在本发明的另一个实施方案中，等渗剂以1mg/ml至7mg/ml的浓度存在。在本发明的另一个实施方案中，等渗剂以8mg/ml至24mg/ml的浓度存在。在本发明的另一个实施方案中，等渗剂以25mg/ml至 50mg/ml的浓度存在。这些特定等渗剂中的每一种构成本发明的替代实施方案。在药物组合物中使用等渗剂是本领域技术人员所熟知的。为方便起见，参考Remington:The Science andPractice of Pharmacy, 19th edition,1995。
在本发明的另一个实施方案中，制剂还包含螯合剂。在本发明的另一个实施方案中，螯合剂选自乙二胺四乙酸(EDTA)，柠檬酸和天冬氨酸的盐，以及它们的混合物。在本发明的另一个实施方案中，螯合剂以0.1mg/ml至5mg/ml的浓度存在。在本发明的另一个实施方案中，螯合剂的浓度为0.1mg/ml至2mg/ml。在本发明的另一个实施方案中，螯合剂以2mg/ml至5mg/ml的浓度存在。这些特定螯合剂中的每一种构成本发明的替代实施方案。在药物组合物中使用螯合剂是本领域技术人员所熟知的。为方便起见，参考Remington:TheScience and Practice of Pharmacy,19th edition,1995。
在本发明的另一个实施方案中，制剂还包含稳定剂。在药物组合物中使用稳定剂是本领域技术人员所熟知的。为方便起见，参考 Remington:The Science and Practiceof Pharmacy,19th edition,1995。
更具体地，本发明的组合物是稳定的液体药物组合物，其治疗活性成分包括在液体药物制剂储存期间可能显示聚集体形成的多肽。“聚集体形成”是指多肽分子之间的物理相互作用，其导致形成可能保持可溶性的寡聚体或从溶液沉淀的大的可见聚集体。“储存期间”是指液体药物组合物或制剂制备后，不立即施用于受试者。相反，在制备之后，将其包装用于以液体形式、冷冻状态或干燥形式储存，以便稍后重建成适合施用于受试者的液体形式或其它形式。“干燥形式”是指液体药物组合物或制剂通过冷冻干燥(即，冻干；例如参见Williams and Polli (1984)J.Parenteral Sci.Technol.38:48-59)，喷雾干燥(参见Masters (1991)in Spray-Drying Handbook(5th ed；Longman Scientific andTechnical,Essez,U.K.),pp.491-676；Broadhead等人(1992)Drug Devel.Ind.Pharm.18:1169-1206；和Mumenthaler等人(1994) Pharm.Res.11:12-20)或空气干燥(Carpenter andCrowe(1988) Cryobiology 25:459-470；and Roser(1991)Biopharm.4:47-53)进行干燥。在储存液体药物组合物期间多肽的聚集体形成可能不利地影响该多肽的生物活性，导致药物组合物的治疗功效丧失。此外，当使用输注系统施用含多肽的药物组合物时，聚集体形成可能引起其它问题，例如管道、膜或泵的堵塞。
本发明的药物组合物可以进一步包含一定量的氨基酸碱，其足以在储存组合物期间降低多肽的聚集体形成。“氨基酸碱”是指氨基酸或氨基酸的组合，其中任何给定的氨基酸以其游离碱形式或其盐形式存在。当使用氨基酸的组合时，所有氨基酸可以以其游离碱形式存在，所有氨基酸可以以其盐形式存在，或一些氨基酸可以以其游离碱形式存在，而其它氨基酸以其盐形式存在。在一些实施方案中，用于制备本发明组合物的氨基酸是携带带电荷侧链的那些，例如精氨酸，赖氨酸，天冬氨酸和谷氨酸。特定氨基酸(例如甘氨酸，甲硫氨酸，组氨酸，咪唑，精氨酸，赖氨酸，异亮氨酸，天冬氨酸，色氨酸，苏氨酸及其混合物)的任何立体异构体(即L，D或DL异构体)或这些立体异构体的组合可以存在于本发明的药物组合物中，只要特定氨基酸以其游离碱形式或其盐形式存在即可。在一些实施方案中，使用L-立体异构体。本发明的组合物也可以用这些氨基酸的类似物配制。“氨基酸类似物”是指天然存在的氨基酸的衍生物，其在存储本发明的液体药物组合物期间产生减少聚集体形成的期望效果。合适的精氨酸类似物包括例如氨基胍，鸟氨酸和N-单乙基L-精氨酸，合适的甲硫氨酸类似物包括乙硫氨酸和丁硫氨酸，合适的半胱氨酸类似物包括S-甲基-L 半胱氨酸。与其它氨基酸一样，氨基酸类似物以其游离碱形式或其盐形式并入组合物中。在本发明的另一个实施方案中，氨基酸或氨基酸类似物以足以防止或延迟蛋白质聚集的浓度使用。
在本发明的另一个实施方案中，可以加入甲硫氨酸(或其它含硫氨基酸或氨基酸类似物)以抑制甲硫氨酸残基氧化成甲硫氨酸亚砜，当作为治疗剂的多肽是包含至少一个易受这样的氧化的甲硫氨酸残基的多肽时。“抑制”是指甲硫氨酸氧化物质随时间的最小积累。抑制甲硫氨酸氧化导致多肽保留在其适当的分子形式。可以使用任何甲硫氨酸的立体异构体(L，D或DL异构体)或其组合。添加量应该足以抑制甲硫氨酸残基的氧化，使得甲硫氨酸亚砜的量可以被管理机构接受。通常，这意味着该组合物含有不超过约10％至约30％的甲硫氨酸亚砜。通常，这可以通过加入甲硫氨酸来实现，使得加入的甲硫氨酸与甲硫氨酸残基的比例范围为约1:1至约1000:1，例如10:1至约100:1。
在本发明的另一个实施方案中，制剂还包含选自高分子量聚合物或低分子化合物的稳定剂。在本发明的另一个实施方案中，稳定剂选自聚乙二醇(例如PEG 3350)，聚乙烯醇(PVA)，聚乙烯吡咯烷酮，羧基-/羟基纤维素或其衍生物(例如HPC，HPC-SL，HPC-L和 HPMC)，环糊精，含硫物质如一硫代甘油，巯基乙酸和2-甲硫基乙醇，以及不同的盐(如氯化钠)。这些特定稳定剂中的每一种构成本发明的替代实施方案。
药物组合物还可以包含另外的稳定剂，其进一步增强其中的治疗活性多肽的稳定性。本发明特别感兴趣的稳定剂包括但不限于保护多肽免受甲硫氨酸氧化的甲硫氨酸和EDTA，以及保护多肽免受与冻融或机械剪切有关的聚集的非离子表面活性剂。
在本发明的另一个实施方案中，制剂还包含表面活性剂。在本发明的另一个实施方案中，表面活性剂选自去垢剂，乙氧基化蓖麻油，聚乙二醇化甘油酯，乙酰化单甘油酯，脱水山梨糖醇脂肪酸酯，聚氧丙烯-聚氧乙烯嵌段聚合物(例如泊洛沙姆，如F68，泊洛沙姆188和407，Triton X-100)，聚氧乙烯脱水山梨糖醇脂肪酸酯，聚氧乙烯和聚乙烯衍生物如烷基化和烷氧基化衍生物(twenens，例如 Tween-20，Tween-40，Tween-80和Brij-35)，单酸甘油酯或其乙氧基化衍生物，甘油二酸酯或其聚氧乙烯衍生物，醇，甘油，凝集素和磷脂(例如磷脂酰丝氨酸，磷脂酰胆碱，磷脂酰乙醇胺，磷脂酰肌醇，二磷脂酰甘油和鞘磷脂)，磷脂衍生物(例如二棕榈酰磷脂酸)和溶血磷脂(例如棕榈酰溶血磷脂酰L-丝氨酸和乙酰胺，胆碱，丝氨酸或苏氨酸的1-酰基-sn-甘油基-3-磷酸酯)和溶血磷脂酰基和磷脂酰胆碱的烷基，烷氧基(烷基酯)，烷氧基(烷基醚)-衍生物，例如溶血磷脂酰胆碱的月桂酰和肉豆蔻酰衍生物，二棕榈酰磷脂酰胆碱和极性头基的修饰，即胆碱，乙醇胺，磷脂酸，丝氨酸，苏氨酸，甘油，肌醇和带正电荷的DODAC，DOTMA，DCP，BISHOP，溶血磷脂酰丝氨酸和溶血磷脂酰苏氨酸，和甘油磷脂(例如头孢氨酸)，甘油糖脂(例如半乳糖吡喃糖苷)，鞘糖脂(例如神经酰胺，神经节苷脂)，十二烷基磷酸胆碱，鸡蛋溶血卵磷脂，夫西地酸衍生物(例如牛磺-二氢福西丁钠等)，长链脂肪酸和其盐C6-C12(例如油酸和辛酸)，酰基肉碱和衍生物，赖氨酸、精氨酸或组氨酸的Nα-酰化衍生物或赖氨酸或精氨酸的侧链酰化衍生物，二肽的Nα-酰化衍生物，其包含赖氨酸，精氨酸或组氨酸和中性或酸性氨基酸的任何组合，三肽的Nα酰化衍生物，其包含中性氨基酸和两个带电荷的氨基酸的组合，DSS(多库酯钠，CAS登记号[577-11-7])，多库酯钙，CAS登记号[128-49-4])，多库酯钾，CAS登记号[7491-09-0])，SDS(十二烷基硫酸钠或十二烷基硫酸钠)，辛酸钠，胆酸或其衍生物，胆汁酸及其盐和甘氨酸或牛磺酸缀合物，熊去氧胆酸，胆酸钠，脱氧胆酸钠，牛磺胆酸钠，甘胆酸钠，N-十六烷基-N,N-二甲基-3-氨基-1-丙磺酸盐，阴离子(烷基- 芳基磺酸盐)单价表面活性剂，两性离子表面活性剂(例如N-烷基-N,N- 二甲基氨基-1-丙磺酸盐，3-胆酰胺-1-丙基二甲基铵-1-丙磺酸盐，阳离子表面活性剂(季铵碱)(例如十六烷基三甲基溴化铵，氯化十六烷基吡啶)，非离子表面活性剂(例如十二烷基β-D-吡喃葡萄糖苷)，泊洛沙胺(例如Tetronic's)，它们是依次添加环氧丙烷和环氧乙烷至乙二胺衍生的四官能嵌段共聚物，或表面活性剂可以选自咪唑啉衍生物或其混合物。这些特定表面活性剂中的每一种都构成本发明的替代实施方案。
在药物组合物中使用表面活性剂是本领域技术人员所熟知的。为方便起见，参考Remington:The Science and Practice of Pharmacy,19th edition,1995。
其它成分可能存在于本发明的肽药物制剂中。这些其它成分可以包括润湿剂，乳化剂，抗氧化剂，填充剂，张力调节剂，螯合剂，金属离子，油性载体，蛋白质(例如，人血清白蛋白，明胶或蛋白质) 和两性离子(例如氨基酸如甜菜碱，牛磺酸，精氨酸，甘氨酸，赖氨酸和组氨酸)。这些其它成分当然不应该不利地影响本发明的药物制剂的整体稳定性。
含有根据本发明的多肽的药物组合物可以在需要这样治疗的患者的几个部位施用，例如在局部部位，例如皮肤和粘膜部位，旁路吸收部位，例如在动脉、静脉、心脏中的施用，以及涉及吸收的部位，例如在皮肤、皮肤下、肌肉或腹部中的施用。
局部施用可能在治疗与局部炎症有关的病况上具有特别的优点，例如治疗与烧伤有关的炎症或与皮肤相关的其它病况。因此，在一些实施方案中，通过局部给药施用。
在一些具体实施方案中，眼滴剂可用于与眼睛相关的病症，例如角膜炎，例如弥漫性层状角膜炎(DLK)。
根据本发明的药物组合物的施用可以通过几种施用途径，例如舌，舌下，颊，口腔，口服，胃和肠中，鼻，肺，例如通过细支气管和肺泡或其组合，表皮，皮肤，透皮，阴道，直肠，眼睛，例如通过结膜，尿道和肠胃外施用给需要这种治疗的患者。
本发明的组合物可以以几种剂型施用，例如作为溶液，悬浮液，乳剂，微乳液，多重乳液，泡沫，药膏，糊剂，膏药，软膏剂，片剂，包衣片剂，漂洗剂，胶囊剂，例如硬明胶胶囊和软明胶胶囊，栓剂，直肠胶囊，滴剂，凝胶，喷雾剂，粉剂，气溶胶，吸入剂，滴眼剂，眼用软膏，眼用漂洗剂，阴道栓，阴道环，阴道软膏，注射溶液，原位转化溶液，例如原位胶凝，原位固化，原位沉淀，原位结晶，输液和植入物。
本发明的组合物可以例如通过共价、疏水和静电相互作用进一步配合或附接至药物载体、药物递送系统和先进药物递送系统，以进一步增强多肽的稳定性，增加生物利用度，增加溶解度，减少不良反应，实现本领域技术人员熟知的慢性治疗，并增加患者依从性或其任何组合。载体、药物递送系统和先进药物递送系统的实例包括但不限于聚合物，例如纤维素和衍生物，多糖，例如葡聚糖和衍生物，淀粉和衍生物，聚(乙烯醇)，丙烯酸酯和甲基丙烯酸酯聚合物，聚乳酸和聚乙醇酸及其嵌段共聚物，聚乙二醇，载体蛋白，例如白蛋白，凝胶，例如热凝胶体系，例如本领域技术人员熟知的嵌段共聚体系，胶束，脂质体，微球，纳米颗粒，病毒样颗粒，细菌样颗粒，液晶及其分散体，L2相及其分散体，它们是本领域技术人员所熟知的脂质-水体系中的相行为，聚合胶束，多重乳液，自乳化，自微乳化，环糊精及其衍生物，以及树枝状大分子。
本发明的组合物可用于使用例如计量吸入器，干粉吸入器和雾化器来制备含有VAR2CSA的多肽的固体，半固体，粉末和溶液用于肺部施用，所有这些都是本领域技术人员所熟知的装置。
本发明的组合物特别可用于配制受控的，持续的，延长的，迟缓的和缓释的药物递送系统。更具体地，但不限于，组合物可用于制备肠胃外受控释放和持续释放系统(两种系统导致施用次数减少多次)，其是本领域技术人员众所周知的。甚至更优选的是皮下施用的受控释放和持续释放系统。不限制本发明的范围，有用的受控释放系统和组合物的实例是水凝胶，油性凝胶，液晶，聚合胶束，微球，纳米颗粒。
用于制备可用于本发明组合物的受控释放系统的方法包括但不限于结晶，缩合，共结晶，沉淀，共沉淀，乳化，分散，高压均化，包封，喷雾干燥，微囊化，凝聚，相分离，溶剂蒸发以产生微球，挤出和超临界流体过程。一般参考Handbook of PharmaceuticalControlled Release(Wise,D.L.,ed.Marcel Dekker,New York,2000)and Drug and thePharmaceutical Sciences vol.99:Protein Formulation and Delivery(MacNally,E.J.,ed.Marcel Dekker,New York,2000)。
肠胃外施用可以通过注射器，任选的笔状注射器通过皮下，肌内，腹膜内或静脉内注射进行。或者，肠胃外施用可以通过输液泵进行。另外的选择是组合物，其可以是用于以鼻或肺部喷雾形式施用根据本发明的优选含有VAR2CSA的多肽的溶液或悬浮液。作为另外的选择，含有本发明多肽的药物组合物也可适用于透皮给药，例如，通过无针注射或来自贴剂，任选地离子电渗贴剂或经粘膜(例如，颊)施用。
术语“稳定的制剂”是指具有增加的物理稳定性，增加的化学稳定性或增加的物理和化学稳定性的制剂。
本文使用的蛋白质制剂的术语“物理稳定性”是指蛋白质形成蛋白质的生物学无活性和/或不溶性聚集体的倾向，这是由于将蛋白质暴露于热机械应力和/或与不稳定的界面和表面例如疏水性表面和界面的相互作用导致的。通过目视检查和/或浊度测量，将填充在合适的容器(例如药筒或小瓶)中的制剂在不同温度下暴露于机械/物理应力(例如搅拌)持续不同时间段来评价含水蛋白质制剂的物理稳定性。制剂的目视检查在具有深色背景的尖锐聚焦光下进行。制剂的浊度的特征在于将浊度的程度排列在例如0至3的刻度上的视觉评分(没有浊度的制剂对应于视觉评分0，并且在日光下显示视觉浊度的制剂对应于视觉评分3)。当在日光下显示视觉浊度时，制剂被分类为关于蛋白质聚集的物理不稳定性。或者，可以通过本领域技术人员熟知的简单浊度测量来评估制剂的浊度。水性蛋白质制剂的物理稳定性也可以通过使用蛋白质的构象状态的光谱试剂或探针来评价。探针优选是优先结合蛋白质的非天然构象异构体的小分子。蛋白质结构的小分子光谱探针的一个实例是硫磺素T。硫磺素T是荧光染料，已广泛用于检测淀粉样蛋白原纤维。在原纤维的存在下，也许还存在其它蛋白质构型，当与原纤维蛋白形式结合时，硫磺素T在约450nm下产生新的激发最大值，并且在约482nm处产生增强的发射。未结合的硫磺素T在这些波长上基本上是非荧光的。
其它小分子可用作蛋白质结构从天然状态到非天然状态的变化的探针。例如优先结合蛋白质的暴露的疏水斑块的“疏水patch”探针。疏水斑块在天然状态下通常被埋蛋白质的三级结构内，但随着蛋白质开始展开或变性而被暴露出来。这些小分子光谱探针的实例是芳香族，疏水性染料，如安瑟沙，吖啶，菲咯啉等。其它光谱探针是金属-氨基酸络合物，例如疏水氨基酸例如苯丙氨酸，亮氨酸，异亮氨酸，甲硫氨酸和缬氨酸等的钴金属络合物。
本文所用的蛋白质制剂的术语“化学稳定性”是指蛋白质结构中的化学共价变化，导致与天然蛋白质结构相比具有潜在的较低生物效力和/或潜在增加的免疫原性的化学降解产物的形成。根据天然蛋白质的类型和性质以及蛋白质暴露的环境可以形成各种化学降解产物。如本领域技术人员所熟知的，在蛋白质制剂的储存和使用过程中，通常可能不能完全避免消除化学降解，并且常常会观察到越来越多的化学降解产物。大多数蛋白质易于脱酰胺，这是其中谷氨酰胺酰基或天冬酰胺酰基残基中的侧链酰胺基团被水解形成游离羧酸的过程。其它降解途径涉及形成高分子量转化产物，其中两个或更多个蛋白质分子通过酰胺化和/或二硫相互作用共价结合，导致形成共价结合的二聚体，低聚物和聚合物降解产物(Stability of Protein Pharmaceuticals,Ahern. T.J.&Manning M.C.,Plenum Press,New York 1992)。可以提及氧化 (例如甲硫氨酸残基)作为化学降解的另一个变型。蛋白质制剂的化学稳定性可以通过在暴露于不同环境条件之后的各个时间点测量化学降解产物的量来评估(降解产物的形成通常可以通过例如升高的温度来加速)。每个单独降解产物的量通常通过使用各种色谱技术(例如 SEC-HPLC和/或RP-HPLC)根据分子大小和/或电荷分离降解产物来确定。
因此，如上所述，“稳定的制剂”是指具有增加的物理稳定性，增加的化学稳定性或增加的物理和化学稳定性的制剂。一般来说，制剂在使用和储存期间必须是稳定的(符合推荐的使用和储存条件)，直到失效期为止。
在本发明的一些实施方案中，包含VAR2CSA多肽的药物制剂在使用超过6周以及储存超过3年时是稳定的。在本发明的其它实施方案中，包含VAR2CSA多肽的药物制剂在使用超过4周以及储存超过 3年时是稳定的。在本发明的另一个实施方案中，包含VAR2CSA多肽的药物制剂在使用超过4周和储存超过两年时是稳定的。在本发明的另一个实施方案中，包含VAR2CSA多肽的药物制剂在使用超过2 周和超过两年时是稳定的。
使用根据本发明的多肽的适应症
根据本发明的优选含有VAR2CSA的多肽可以用于与CSA的表达例如不适当表达相关的广泛适应症，例如在各种癌症中，例如转移性癌症包括黑素瘤，如C32黑素瘤，肉瘤，肺癌，少突胶质细胞瘤，人脑肿瘤，包括神经胶质瘤，白血病，例如淋巴细胞白血病和急性骨髓性白血病，和癌，例如鳞状细胞癌和乳腺癌，肾细胞癌，软骨肉瘤和胰腺细胞癌。多肽也可以用于癌症干细胞，因此在发展成癌症前靶向细胞。与CSA的表达例如不适当表达相关的其它病况是软骨的病况和/或瘢痕组织的发展。
本发明的具体实施方案
如本文所述，本发明涉及多肽，其包含i)第一多肽结构域，其是嵌合抗原受体(CAR)的跨膜结构域和胞内结构域，以及ii)选自 VAR2CSA多肽或分割蛋白结合系统的肽部分的第二胞外多肽结构域，以及多肽其包含i)第一多肽结构域，其是VAR2CSA多肽或其它肿瘤靶向剂，例如单链scFv，和ii)选自分割蛋白结合系统的肽部分、单结构域抗体例如scFv或纳米抗体中的一个或多个的第二多肽结构域。
在一些实施方案中，包含所述CAR和分割蛋白结合系统的肽部分的多肽可以在体外或体内与分割蛋白标记的肿瘤靶向剂例如抗体组合。
在一些实施方案中，分割蛋白结合系统的一个肽部分选自K-Tag (SEQ ID NO：132)，SpyCatcher(SEQ ID NO：129)，SpyCatcher -ΔN(SEQ ID NO：136)，SpyTag(SEQ IDNO：130)，最小Spytag 序列(SEQ ID NO：131)，split-Spy0128(SEQ ID NO：135)，异肽Spy0128(SEQ ID NO：134)或其任何反向序列或其具有至少约 80％，例如至少约82,84,86,88,90,92,94,96,98或99％的序列同一性的变体。
在一些实施方案中，多肽还包含一个或多个接头或铰链序列，例如GC接头。
在一些实施方案中，VAR2CSA多肽由SEQ ID NO：55或SEQ ID NO：56或其具有结合硫酸软骨素A(CSA)的能力的片段或变体组成，所述硫酸软骨素A(CSA)可以是被呈现在蛋白聚糖(CSPG) 上的。
在一些实施方案中，VAR2CSA多肽是VAR2CSA的片段，其由以下的连续氨基酸序列组成：
a)ID1，和
b)DBL2Xb，以及任选地
c)ID2a。
在一些实施方案中，VAR2CSA多肽以通过小于100nM，例如小于80nM，例如小于70nM，例如小于60nM，例如小于50nM，例如小于40nM，例如小于30nM，例如小于26nM，例如小于24nM，小于22nM，例如小于20nM，例如小于18nM，例如小于16nM，例如小于14nM，例如小于12nM，例如小于10nM，例如小于9nM，例如小于8nM，例如小于7nM，例如小于6nM，或小于4nM的KD测量的亲和力结合蛋白聚糖(CSPG)上的硫酸软骨素A(CSA)。
在一些实施方案中，VAR2CSA多肽包含与SEQ ID NO：1的 1-577，SEQ ID NO：3的1-592，SEQ ID NO：4的1-579，SEQ ID NO： 5的1-576，SEQ ID NO：10的1-586，SEQ ID NO：11的1-579，SEQ ID NO：29的1-565，SEQ ID NO：34的1-584，SEQ ID NO：36的 1-569，SEQ ID NO：37的1-575，SEQ ID NO：38的1-592，SEQ ID NO：41的1-603，SEQ ID NO：43的1-588，SEQ IDNO：44的1-565， SEQ ID NO：45的1-589，SEQ ID NO：48的1-573，SEQ ID NO： 53的1-583，SEQ ID NO：54的1-569或SEQ ID NO：128的1-640 的任何一个氨基酸序列具有至少70％序列同一性的氨基酸序列。
在一些实施方案中，VAR2CSA多肽包含与SEQ ID NO：1的 578-640，SEQ ID NO：3的593-656，SEQ ID NO：4的580-643，SEQ ID NO：5的577-640，SEQ ID NO：10的587-650，SEQID NO：11 的580-643，SEQ ID NO：29的566-628，SEQ ID NO：34的585-647， SEQ ID NO：36的570-632，SEQ ID NO：37的576-639，SEQ ID NO： 38的593-655，SEQ ID NO：41的604-667，SEQ ID NO：43的589-652， SEQ ID NO：44的566-628，SEQ ID NO：45的590-653，SEQ ID NO：48的574-637，SEQ ID NO：53的584-646或SEQ ID NO：54的570-632 的氨基酸序列具有至少70％序列同一性的氨基酸序列。
在一些实施方案中，VAR2CSA多肽包含与SEQ ID NO：2,6,8,9, 12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,30,31,32, 33,35,39,40,42,46,47,49,50,51,52,或128的氨基酸序列具有至少 70％序列同一性的氨基酸序列。
在一些实施方案中，VAR2CSA多肽由与SEQ ID NO：1的1-577，SEQ ID NO：3的1-592，SEQ ID NO：4的1-579，SEQ ID NO：5 的1-576，SEQ ID NO：10的1-586，SEQ ID NO：11的1-579，SEQ ID NO：29的1-565，SEQ ID NO：34的1-584，SEQ ID NO：36的 1-569，SEQ ID NO：37的1-575，SEQ ID NO：38的1-592，SEQ ID NO：41的1-603，SEQ ID NO：43的1-588，SEQ IDNO：44的1-565， SEQ ID NO：45的1-589，SEQ ID NO：48的1-573，SEQ ID NO： 53的1-583，SEQ ID NO：54的1-569或SEQ ID NO：128的1-640 的任何一个氨基酸序列具有至少70％序列同一性的氨基酸序列组成。
在一些实施方案中，VAR2CSA多肽由选自SEQ ID NO：1,3-5,10, 11,29,34,36-38,41,43-45,48,53,54和128的氨基酸序列组成。
在一些实施方案中，VAR2CSA多肽由长度小于700个氨基酸，例如小于690个氨基酸，例如小于680个氨基酸，例如小于670个氨基酸，例如小于660个氨基酸，例如小于650个氨基酸，例如小于640 个氨基酸，例如小于630个氨基酸，例如小于620个氨基酸，例如小于610个氨基酸，例如小于600个氨基酸，例如小于590个氨基酸，例如小于580个氨基酸，例如小于570个氨基酸的氨基酸序列组成。
在一些实施方案中，VAR2CSA多肽具有在非还原条件下在 SDS-PAGE上的小于约100KDa的分子量。
在一些实施方案中，VAR2CSA多肽是非糖基化的。
在一些实施方案中，嵌合抗原受体(CAR)的所述跨膜结构域和 /或胞内结构域来源于第1代、第2代或第3代CAR。
在一些实施方案中，包含嵌合抗原受体(CAR)的跨膜结构域和胞内结构域作为第一多肽结构域的所述多肽还包含其它调节元件或自杀基因系统，例如诱导型半胱天冬酶-9(iCASP9)或睡美人(SB)转座子系统。
在一些实施方案中，嵌合抗原受体(CAR)的所述跨膜结构域和 /或胞内结构域包含CD28和/或CD3ζ信号转导结构域。
在一些实施方案中，嵌合抗原受体(CAR)的所述跨膜结构域和 /或胞内结构域是如WO2014127261，WO13126729，WO13126733， WO14055442，WO14055771，WO2014039523，WO14179759和 WO14138704中任一个所描述的。
序列，包括VAR2CSA多肽的序列：
>fcr3 745个氨基酸|640 aa；下划线序列对应于FCR3的ID1结构域，粗体字序列对应于FCR3的DBL2Xb结构域。剩余序列是ID2a(SEQ ID NO：1)
NYIKGDPYFAEYATKLSFILNPSDANNPSGETANHNDEACNCNESGISSVGQAQTSGPSSNKTCITHSSIKTNKKKE CKDVKLGVRENDKDLKICVIEDTSLSGVDNCCCQDLLGILQENCSDNKRGSSSNDSCDNKNQDECQKKLEKVFASLT NGYKCDKCKSG TSRSKKKWIWKKSSGNEEGLQEEYANTIGLPPRTQSLYLGNLPKLENVCEDVKDINFDTKEKFLAGCLIVSFHEGKNLKKRYPQNKNSGNKENLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQNNFGKLFGKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWTAMKHGAEMNITTCNADGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVENFCEQRQAKVKDVITNCKSCKESGNKCKTECKTKCKDECEKYKKFIEACGTAGGGIGTAGSPWSKRWDQIYKRYSKHIEDAKRNRKAGTKNCGTSSTTNAAASTDENKCVQSDIDSFFKHLIDIGLTTPSSYLSNVLDDNICGADKAPWTTYTTYTTTEKCNKERDKSKSQSSDTLVVVNVPSPLGNTPYRYKYACQCKIPTNEETCDDRKEYMNQWSCGSARTMKRGYKNDNYELCKYNGVDVKPTTVRSNSSKLD
>gi|254952610|gb|ACT97135.1|VAR2CSA[恶性疟原虫]|341aa(SEQ ID NO:2)
KCDKCKSGTSRSRKIWTWRKSSGNKEGLQEEYANTIGLSPRTQLLYLGNLRKLENVCEDVTDINFDTKEKFLAGCLIAAFHEGKNLKKRYLEKKKGDNNSKLCKDLKYSFADYGDLIKGTSIWDNDFTKDLELNLQQIFGKLFRKYIKKKNISTEQDTSYSSLDELRESWWNTNKKYIWLAMKHGAGMNSTTCSCSGDSSSGENQTNSCDDIPTIDLIPQYLRFLQEWVEHFCEQRQAKVKDVITNCNSCKESGGTCNSDCEKKCKNKCDAYKTFIEDCKGVGGTGTAGSSWVKRWYQIYMRYSKYIEDAKRNRKAGTKSCGT SSTTNVSVSTDENKCVQS-
>M24 745个氨基酸|656aa(SEQ ID NO:3)
DYIKGDPYFAEYATKLSFILNSSDANNPSGETANHNDEVCNPNESEISSVGQAQTSDPSSNKTCNTHSSIKANKKKVCKHVKLGINNNDKVLRVCVIEDTSLSGVENCCFKDLLGILQENCSDNKSGSSSNGSCNNKNQEACEKNLEKVLASLTNCYKCDKCKSGTSTVNKNWIWKKSSGNKEGLQKEYANTIGLPPRTHSLYLGNLPKLENVCEDVKDINFDTKEKFLAGCLIAAFHEGKNLKKRYPQNKNDDNNSKLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQQIFGKLFRKYIKKNISTEQDTLYSSLDELRESWWNTNKKYIWLAMKHGAGMNITTCCGDGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVEHFCKQRQEKVKDVINSCNSCKNTSSKTKLGDTCNSDCEKKCKIECEKYKKFIEECRTAVGGTAGSSWSKRWDQIYKMYSKHIEDAKRNRKAGTKNCGITTGTISGESSGANSGVTTTENKCVQSDIDSFFKHLIDIGLTTPSSYLSIVLDDNICGDDKAPWTTYTTYTTYTTTEKCNKERDKSKSQQSNTSVVVNVPSPLGNTPHGYKYACQCKIPTNEETCDDRKEYMNQWISDTSKNPKGSGSTNNDYELYTYNGVKETKLPKKLNSP KLD
>KMWII 745个氨基酸|643aa(SEQ ID NO:4)
DYIKDDPYSKEYTTKLSFILNSSDANTSSGETANHNDEACNCNESEISSVGQAQTSGPSSNKTCITHSFIKANKKKVCKDVKLGVRENDKVLRVCVIEDTSLSGVDNCCCQDLLGILQENCSDNKRGSSSNGSCNNKNQDECQKKLEKVFVSLTNGYKCDKCKSGTSTVNKKWIWKKSSGNEKGLQKEYANTIGLPPRTQSLYLGNLPKLGNVCEDVTDINFDTKEKFLAGCLIAAFHEGKNLKISHEKKKGDNGKKLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQKAFGKLFGKYIKKNIASDENTSYSSLDELRESWWNTNKKYIWTAMKHGAEMNSTMCNADGSVTGSGSSCDDIPTTDFIPQYLRFLQEWVEHFCKQRQEKVNAVIENCNSCKNTSGERKIGGTCNGDCKTECKNKCEAYKNFIEDCKGGDGTAGSSWVKRWDQIYKRYSKHIEDAKRNRKAGTKSCGPSSITNASVSTDENKCVQSDIDSFFKHLIDIGLTTPSSYLSIVLDENNCGEDNAPWTTYTTYTTTEKCNKDKKKSKSQSCNTAVVVNVPSPLGNTPHEYKYACQCKIPTTEETCDDRKEYMNQWISDTSKKQKGSGSTNNDYELYTYTGVKETKLPKKLNSPKLD
>1248 745个氨基酸|640aa(SEQ ID NO:5)
SYVKNDPYSKEYVTKLSFILNPSDANNPSGETANHNDEACNPNESEIASVGQAQTSDRLSQKACITHSFIGANKKIVCKDVKLGVREKDKDLKICVIEDDSLRGVENCCFKDLLGILQENCSDNKSGSSSNGSCNNKNQDECQKKLDEALASLHNGYKCDKCKSGTSRSKKIWTWRKFPGNGEGLQKEYANTIGLPPRTQSLYLGNLRKLENVCKGVTDINFDTKEKFLAGCLIAAFHEGKNLKISNKKKNDDNGKKLCKDLKYSFADYGDLIKGTSIWDNEYTKDLELNLQKIFGKLFRKYIKKNIASDENTLYSSLDELRESWWNTNKKYIWLAMKHGTTCSSGSGDNGDGSVTGSGSSCDDMSTIDLIPQYLRFLQEWVEHFCKQRQEKVKDVIENCKSCKNTSGERIIGGTCGSDCKTKCKGECDAYKNFIEECKRGDGTAGSPWSKRWDQIYMRYSKYIEDAKRNRKAGTKNCGTSSTTNAAENKCVQSDIDSFFKHLIDIGLTTPSSYLSIVLDENICGDDKAPWTTYTTYTTTEKCNKETDKSKSQSCNTAVVVNVPSPLGNTPHGYKYACECKIPTTEETCDDRKEYMNQWISDTSKKPKGGRSTNNDYELYTYNGVKETKLPKKSSSSKLD
>gi|254952618|gb|ACT97139.1|VAR2CSA[恶性疟原虫]|358aa(SEQ ID NO:6)
KCEKCKSEQSKKNNNIWIWRKFPGNGEGLQKEYANTIGLPPRTHSLYLGNLPKLENVCKDVKDINFDTKEKFLAGCLIAAFHEGKNLKTTYPQNKNADNNSKLCKDLKYSFADYGDLIKGTSIWDNDFTKDLELNLQKIFGKLFRKYIKKNIASDENTLYSSLDELRESWWNTNKKYIWLAMKHGAEMNSTMCNGDGSVTGSSDSGSTTCSGDNGSISCDDIPTIDLIPQYLRFLQEWVEHFCKQRQEKVKPVIENCKSCKNTSGERIIGGTCGSDCEKKCKGECDAYKKFIEECKGGGGGTGTAGSPWSKRWDQIYKRYSKYIEDAKRNRKAGTKSCGPSSTTNAAASTTESKCVQS
>gi|254952592|gb|ACT97126.1|VAR2CSA[恶性疟原虫]|333aa(SEQ ID NO:7)
KCDKCKSEQSKKNNKNWIWKQFPGNGEGLQKEYANTIGLPPRTHSLYLGNLPKLENVCKGVTDINFDTKEKFLAGCLIAAFHEGKNLKTSHEKKKGDNGKKLCKDLKYSFADYGDLIKGTSIWDNDFTKDLELNLQQIFGKLFRKYIKKNISAEQDTSYSSLDELRESWWNTNKKYIWLAMKHGTTCSSGSGDNGDGSVTGSGSSCDDMPTTDFIPQYLRFLQEWVEHFCKQRQEKVNAVITNCKSCKESGGTCNSDCEKKCKDECEKYKKFIEECRTAADGTAGSSWSKRWDQIYKMYSKHIEDAKRNRKAGTKNCGTSS TTNAAENKCVQS
>gi|90193467|gb|ABD92329.1|红细胞膜蛋白1[恶性疟原虫]|269aa(SEQ ID NO:8)
DYIKDDPYSKEYTTKLSFILNSSDANTSSGETANHNDEACNCNESEIASVEQASISDRSSQKAYITHSSIKTNKKKVCKYVKLGINNNDKVLRVCVIEDTSLSGVENCCFKDLLGILQENCSDNKRGSSFNDSCNNNNEEACQKKLEKVLASLTNGYKCEKCKSGTSRSKKKWIWKKSSGKEGGLQKEYANTIGLPPRTQSLYLGNLPKLENVCKGVTDINFDTKEKFLAGCLIAAFHEGKNLKPSHQNKNDDNNSKLCKDLKYSFADY
>gi|254952616|gb|ACT97138.1|VAR2CSA[恶性疟原虫]|333aa(SEQ ID NO:9)
KCDKCKSGTSRSKKKWTWRKSSGNKEGLQKEYANTIGLPPRTHSLYLGNLRKLENVCEDVTDINFDTKEKFLAGCLIAAFHEGKNLKTTYPQNKNDDNNSKLCKALKYSFADYGDLIKGTSIWDNDFTKDLELNLQKIFGKLFRKYIKKNISTEQHTSYSSLDELRESWWNTNKKYIWLAMKHGAEMNGTTCSCSGDSSDDIPTIDLIPQYLRFLQEWVEHFCKQRQAKVNAVINSCNSCKNTSGERKLGGTCGSECKTECKNKCDAYKEFIDGTGSGGGTGTAGSSWVKRWDQIYKRYSKYIEDAKRNRKAGSKNCGTSSTTN AAESKCVQS
>hb31 745个氨基酸|650aa(SEQ ID NO:10)
SYVKNNPYSAEYVTKLSFILNSSDANTSSETPSKYYDEVCNCNESEISSVGQAQTSGPSSNKTCITHSSIKTNKKKVCKDVKLGINNNDKVLRVCVIEDTSLSGVDNCCCQDLLGILQENCSDKNQSGSSSNGSCNNKNQDECQKKLEKVFASLTNGYKCDKCKSGTSRSKKKWIWRKSSGNEEGLQKEYANTIGLPPRTQSLYLGNLRKLENVCKGVTDINFDTKEKFLAGCLIAAFHEGKNLKTTYPQNKKKLCKDLKYSFADYGDLIKGTSIWDNEYTKDLELNLQKAFGKLFRKYIKKNISTEQHTLYSSLDELRESWWNTNKKYIWLAMKHGAGMNSTTCCGDGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVEHFCKQRQEKVNAVIENCNSCKECGDTCNGECKTECEKKCKIECEKYKTFIEECVTAVGGTSGSPWSKRWDQIYKRYSKYIEDAKRNRKAGTKNCGITTGTISGESSGANSGVTTTENKCVQSDIDSFFKHLIDIGLTTPSSYLSIVLDDNICGADNAPWTTYTTYTTYTTTKNCDIKKKTPKSQPINTSVVVNVPSPLGNTPHGYKYACQCKIPTTEESCDDRKEYMNQWIIDTSKKQKGSGSTNNDYELYTYNGVKETKLPKKSSSSKLD
>hb32 745个氨基酸|643aa(SEQ ID NO:11)
SYVKDDPYSAEYVTKLSFILNSSDANTSSETPSKYYDEVCNCNESEISSVGQAQTSGPSSNKTCITHSSIKTNKKKVCKDVKLGINNNDKVLRVCVIEDTSLSGVDNCCCQDLLGILQENCSDKNQSGSSSNGSCNNKNQDECQKKLEKVFASLTNGYKCDKCKSGTSRSKKKWIWRKSSGNEEGLQKEYANTIGLPPRTQSLYLGNLPKLENVCKGVTDIIYDTKEKFLSGCLIAAFHEGKNLKTSHEKKNDDNGKKLCKALEYSFADYGDLIKGTSIWDNDFTKDLELNLQKIFGKLFRKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWTAMKHGAGMNSTTCSGDGSVTGSGSSCDDMPTIDLIPQYLRFLQEWVEHFCKQRQEKVKDVITNCNSCKECGDTCNGECKTECKTKCKGECEKYKNFIEECNGTADGGTSGSSWSKRWDQIYKRYSKYIEDAKRNRKAGTKNCGTSSTTNAAASTTENKCVQSDIDSFFKHLIDIGLTTPSSYLSNVLDDNICGEDKAPWTTYTTYTTKNCDIQKKTPKPQSCDTLVVVNVPSPLGNTPHGYKYVCECKIPTTEETCDDRKEYMNQWIIDTSKKQKGSGSTNNDYELYTYNGVQIKQAAGTLKNSKLD
>gi|90193475|gb|ABD92333.1|红细胞膜蛋白1[恶性疟原虫]|269aa(SEQ ID NO:12)
NYIKGDPYSAEYATKLSFILNSSDTENASEKIQKNNDEVCNCNESEIASVEQAPISDRSSQKACITHSSIKANKKKVCKHVKLGVRENDKDLKICVIEDTSLSGVDNCCCQDLLGILQENCSDNKSGSSSNGSCNNNNEEICQKKLEKVLASLTNGYKCDKCKSGTSTVNKNWIWKKYSGKEGGLQEEYANTIGLPPRTQSLYLGNLPKLENVCEDVKDINFDTKEKFLAGCLIAAFHEGKNLKTSNKKKNDDNNSKLCKALKYSFADY
>gi|254952600|gb|ACT97130.1|VAR2CSA[恶性疟原虫]|344aa(SEQ ID NO:13)
KCDKCKSGTSTVNKKWIWKKYSGTEGGLQEEYANTIALPPRTQSLYLGNLPKLENVCKDVTDINFDTKEKFLAGCLIAAFHEGKNLKTTYLEKKKGDNGKKNDDNNSKLCKALKYSFADYGDLIKGTSIWDNDFTKDLELNLQQIFGKLFRKYIKKNIASDENTLYSSLDELRESWWNTNKKYIWLAMKHGAGMNSTMCNADGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVEHFCKQRQAKVKDVITNCNSCKECGGTCNGECKTECEKKCKGECDAYKKFIEECKGKADEGTSGSSWSKRWDQIYKRYSKYIEDAKRNRKAGTKNCGPSSTTSTAESKCVQS
>gi|254952598|gb|ACT97129.1|VAR2CSA[恶性疟原虫]|334aa(SEQ ID NO:14)
KCDKCKSEQSKKNNNIWIWKKSSGTEGGLQKEYANTIALPPRTQSLYLGNLRKLENVCEDVKDINFDTKEKFLAGCLIAAFHEGKNLKKRYLEKKNGDNNSKLCKALKYSFADYGDLIKGTSIWDNEYTKDLELNLQKIFGKLFRKYIKKNNTAEQHTSYSSLDELRESWWNTNKKYIWLAMKHGTTCSSGSGDNGSISCDDIPTIDLIPQYLRFLQEWVEHFCEQRQGKVNAVIENCNSCKNTSSKTKLGGTCNGECKTECKGECDAYKEFIEKCKGTAAEGTSGSSWVKRWYQIYMRYSKYIEDAKRNRKAGTKNCGTSSTT STAESKCVQS
>gi|254952596|gb|ACT97128.1|VAR2CSA[恶性疟原虫]|332aa(SEQ ID NO:15)
KCDKCKSEQSKKNNNIWIWKKSSGTEGGLQKEYANTIALPPRTQSLYLGNLRKLENVCEDVKDINFDTKEKFLAGCLIAAFHEGKNLKKRYLEKKNGDNNSKLCKALKYSFADYGDLIKGTSIWDNEYTKDLELNLQKIFGKLFRKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWTAMKHGTTCSSGSGDNGSISCDDIPTIDLIPQYLRFLQEWVEHFCEQRQEKVKDVIKNCNSCKECGGTCNGECKTECKNKCKDECDAYKKFIEECEGKAAEGTSGSSWSKRWDQIYKRYSKYIEDAKRNRKAGTKNCGTSSTTS TAENKCVQS
>gi|90193465|gb|ABD92328.1|红细胞膜蛋白1[恶性疟原虫]|267aa(SEQ ID NO:16)
NYIKDDPYSAEYTTKLSFILNSSDTENASEKIQKNNDEVCNPNESGIACVELAQTSGSSSNKTCNTHSFIKANKKKVCKDVKLGINKKDKDLKICVIEDDSLRGVDNCCCQDLLGILQENCSDKNQSGSSSNGSCNNKNQEACQKKLENVFASLTNGYKCEKCKSEQSKKNNKNWIWKKYSVKEEGLQKEYANTIALPPRTQSLYLGNLPKLGNVCKGVTDINFDTKEKFLAGCLIAAFHEGKNLKTTYLQNKKKLCKALKYSFADY
>gi|90193477|gb|ABD92334.1|红细胞膜蛋白1[恶性疟原虫]|263aa(SEQ ID NO:17)
DYIKGDPYFAEYATKLSFILNSSDANTSSGETANHNDEACNPNESEIASVEQASISDRSSQKACNTHSSIKANKKKECKHVKLGVRENDKDLKICVIEDTSLSGVDNCCCQDLLGILQENCSDNKRGSSSNGSCDKNSEEICQKKLDEALASLHNGYKNQKCKSEQSKKNKNKWIWKKSSGNEKGLQKEYANTIGLPPRTQSLYLGNLPKLENVCEDVTDINFDTKEKFLAGCLIAAFHEGKNLKTTYPQNKNDDNGKKLCKD
>gi|254952594|gb|ACT97127.1|VAR2CSA[恶性疟原虫]|338aa(SEQ ID NO:18)
KCDKCKSEQSKKNNNIWIWKKSSGNKKGLQKEYANTIGLPPRTQSLYLGNLPKLENVCKDVTDINFDTKEKFLAGCLIAAFHEGKNLKISNEKKNDDNGKKLCKDLKYSFADYGDLIKGTSIWDNEYTKDLELNLQNNFGKLFRKYIKKNNTAEQHTLYSSLDELRESWWNTNKKYIWLAMKHGTTCSSGSGDNGDGSVTGSGSSCDDMSTIDLIPQYLRFLQEWVEHFCKQRQEKVNAVIENCNSCKNTSSKTKLGGTCNGECKTECEKKCKDECEKYKEFIEECKRGDGTAGSPWVKRWDQIYMRYSKYIEDAKRNRKA GTKSCGTSAAENKCVQS
>gi|254952602|gb|ACT97131.1|VAR2CSA[恶性疟原虫]|341aa(SEQ ID NO:19)
KCDKCKSEQSKKNNNIWIWKKSSGDEKGLQKEYANTIALPPRTQSLYLGNLPKLENVCKDVTDINFDTKEKFLAGCLIAAFHEGKNLKTSHQNKNADNGKKNDDNGKKLCKALKYSFADYGDLIKGTSIWDNEYTKDLELNLQQIFGKLFRKYIKRNNTAEQHTLYSSLDELRESWWNTNKKYIWLAMKHGTTCSSGSGDNGDGSVTGSGSSCDDMSTIDLIPQYLRFLQEWVEHFCKQRQEKVKDVITNCNSCKECGGTCGSDCKTKCEAYKKFIEECNGTADGGTSGSSWSKRWDQIYKRYSKYIEDAKRNRKAGTKNC GPSSGANSGVTTTENKCVQS
>gi|254952660|gb|ACT97160.1|VAR2CSA[恶性疟原虫]|352aa(SEQ ID NO:20)
KCEKCESEQSKKNNKYWIWKKSSGNGEGLQEEYANTIALPPRTHSLCLVCLHEKEGKKTQELKNIRTNSELLKERIIAAFHEGKNLKTSPQNKNDNGKKLCKDLKYSFADYGDLIKGTSIWDNEYTKDLELNLQKIFGKLFRKYIKKNNTAEQHTLYSSLDELRESWWNTNKKYIWLAMKHGAGMNSTMCNADGSVTGSSDSGSTTCCGDNGSISCDDMPTIDLIPQYLRFLQEWVEHFCEQRQEKVNAVITNCKSCKECGGTCNSDCEKKCKAYKEFIEKCKGGGTEGTSGSSWSKRWDQIYKRHSKHIEDAKRNRKAGTKNCGITTGTISGESSGANSGVTTTENKCVQS
>gi|254952652|gb|ACT97156.1|VAR2CSA[恶性疟原虫]|344aa(SEQ ID NO:21)
KCDKCKSGTSRSRKIWTWRKFRGNGEGLQKEYANTIGLSPRTQLLYLVCLHEKGKKTQELKNISTNSELLKEWIIAAFHEGKNLKTTYPQKKNDDNGKKLCKALKYSFADYGDLIKGTSIWDNDFTKDLELNLQKIFGKLFRKYIKKNIASDENTSYSSLDELRESWWNTNKKYIWTAMKHGAGMNGTTCCGDGSVTGSSDSGSTTCCGDGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVEHFCEQRQEKVKDVITNCKSCKESEKKCKNKCDAYKEFIDGTGSGGGTGTAGSSWSKRWDQIYMRYSKYIEDAKRNRKAGTKN CGTSSGANSGVTTTENKCVQS
>gi|254952622|gb|ACT97141.1|VAR2CSA[恶性疟原虫]|350aa(SEQ ID NO:22)
KCEKCKSEQSKKNNKIWTWRKFPGNGEGLQKEYANTIGLSPRTQLLYLVCLHEKGKKTQHKTISTNSELLKEWIIAAFHEGKNLKKRYLEKKKGDNNSKLCKDLKYSFADYGDLIKGTSIWDNDFTKDLELNLQQIFGKLFRKYIKKNIASDENTSYSSLDELRESWWNTNKKYIWTAMKHGAGMNSTMCNGDGSVTGSSDSGSTTCSGDNGSISCDDIPTIDLIPQYLRFLQEWVEHFCEQRQEKVKDVIKNCNSCKECGGTCNGECKTECKNKCKDECEKYKNFIEVCTGGDGTAGSPWSKRWYQIYMRYSKYIEDAKRNRKAGTKSCGTSSGANSGVTTTESKCVQS
>gi|254952626|gb|ACT97143.1|VAR2CSA[恶性疟原虫]|359aa(SEQ ID NO:23)
KCEKCKSEQSKKNNKNWIWRKFPGNGEGLQKEYANTIGLPPRTHSLYLVCLHEKGKKTQELKNIRTNSELLKEWIIAAFHEGKNLKKRYHQNNNSGNKKKLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQQIFGKLFRKYIKKNISTEQDTLYSSLDELRESWWNTNKKYIWLAMKHGAGMNSTTCCGDGSVTGSSDSGSTTCSGDNGSISCDDMPTIDLIPQYLRFLQEWVEHFCEQRQEKVKDVIENCKSCKNTSGERIIGGTCNGECKTECEKKCKAACEAYKTFIEECEGKAAEGTSGSSWSKRWYQIYMRYSKYIEDAKRNRKAGTKNCGKSSGANSGVTTTENKCVQS
>gi|90193469|gb|ABD92330.1|红细胞膜蛋白1[恶性疟原虫]|270aa(SEQ ID NO:24)
NYIKDDPYSKEYVTKLSFIPNSSDANNPSGETANHNDEVCNPNESEISSVEHAQTSVLLSQKAYITHSSIKANKKKVCKYVKLGVRENDKDLKICVIEDDSLRGVENCCFKDFLRILQENCSDNKRESSSNGSCNNNNEEACEKNLDEALASLTNCYKNQKCKSGTSTVNNNKWIWKKSSGKEGGLQKEYANTIGLPPRTQSLCLVVCLDEKEGKTQELKNIRTNSELLKEWIIAAFHEGKNLKKRYHQNKNDDNNSKLCKALKYSFADY
>gi|254952644|gb|ACT97152.1|VAR2CSA[恶性疟原虫]|334aa(SEQ ID NO:25)
KCDKCKSEQSKKNNKYWIWKKYSVKEGGLQKEYANTIALPPRTQSLCLVVCLDEKEGKTQELKNIRTNSELLKERIIAAFHEGKNLKTYHEKKKGDDGKKLCKDLKYSFADYGDLIKGTSIWDNDFTKDLELNLQKIFGKLFRKYIKKNNTAEQHTSYSSLDELRESWWNTNKKYIWTAMKHGAEMNGTTCSCSGDSSNDIPTIDLIPQYLRFLQEWVEHFCEQRQAKVNAVIKNCKSCKECGGTCNGECKTECKTKCKGECEKYKEFIEKCEGQAAEGTSGSSWSKRWYQIYMRYSKYIEDAKRNRKAGTKNCGTSSGA NSGVTTTENKCVQS
>gi|254952642|gb|ACT97151.1|VAR2CSA[恶性疟原虫]|351aa(SEQ ID NO:26)
KCDKCKSEQSKKNNKNWIWKKYSGTEGGLQKEYANTIALPPRTQSLYLVCLHEKEEKTQELKNISTNSELLKEWIIAAFHEGKNLKISPQNKNDNGKNLCKDLKYSFADYGDLIKGTSIWDNDFTKDLELNLQQIFGKLFRKYIKKNNTAEQDTLYSSLDELRESWWNTNKKYIWTAMKHGAGMNGTTCCGDGSVTGSSDSGSTTCCGDGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVEHFCEQRQAKVKDVIKNCNSCKECGGTCNGECKTECEKKCKGECEAYKKFIEKCNGGGGEGTSGSSWSKRWDQIYMRYSKYIEDAKRNRKAGTKNCGTSSTTNAAENKCVQS
>gi|254952658|gb|ACT97159.1|VAR2CSA[恶性疟原虫]|353aa(SEQ ID NO:27)
KCDKCKSGTSTVNKKWIWKKFPGKEGGLQEEYANTIALPPRTQSLCLVVCLDEKEGKTQHKTISTNSELLKEWIIAAFHEGKNLKISNKKKNDENNSKLCKDLKYSFADYGDLIKGTSIWDNDFTKDLELNLQKIFGKLFRKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWLAMKHGTTCSSGSGDNGDGSVTGSSDSGSTTCCGDGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVEHFCKQRQAKVKDVIENCKSCKNTSSKTKLGDTCNSDCKTKCKVACEKYKEFIEKCVSAAGGTSGSSWVKRWDQIYMRYSKYIEDAKRNRKAGTKNCGPSSTTSTAESKCVQS
>gi|254952640|gb|ACT97150.1|VAR2CSA[恶性疟原虫]|327aa(SEQ ID NO:28)
KCDKCKSGTSTVNKKWIWKKYSGKEGGLQKEYANTIGLPPRTQSLCLVCLHEKEGKTQELKNISTNSELLKEWIIAAFHEGKNLKISNKKKNDDNGKKLCKDLKYSFADYGDLIKGTSIWDNDFTKDLELNLQKIFGKLFRKYIKKNNTAEQDTLYSSLDELRESWWNTNKKYIWTAMKHGAGMNSTTCSCSGDSSNDIPTIDLIPQYLRFLQEWVEHFCKQRQEKVNAVITNCKSCKESGGTCNSDCEKKCKIECEKYKNFIEKCVTAAGGTSGSSWSKRWDQIYKMYSKYIEDAKRNRKAGTKNCGPSSTTNAAASTDENK CVQS
>dd2full 745个氨基酸|628aa(SEQ ID NO:29)
NYIKGDPYFAEYATKLSFILNSSDTENASETPSKYYDEACNCNESEIASVGQAQTSGPSSNKTCITHSSIKTNKKKECKDVKLGINNNDKVLRVCVIEDTSLSGVDNCCCQDLLGILQENCSDNKRGSSSNGSCDKNSEEICQKKLEKVFASLTNGYKCDKCKSGTSRSKKKWIWKKSSGNEEGLQKEYANTIGLPPRTQSLCLVCLHEKEGKTQHKTISTNSELLKEWIIAAFHEGKNLKTSHEKKNDDNGKKLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQKIFGKLFRKYIKKNNTAEQHTSYSSLDELRESWWNTNKKYIWTAMKHGAGMNGTTCSCSGDSSNDMPTIDLIPQYLRFLQEWVEHFCKQRQEKVNAVIENCNSCKESGGTCNSDCKTECKNKCEAYKEFIEDCKGGGTGTAGSPWSKRWDQIYKRYSKHIEDAKRNRKAGTKNCGTSSTTNAAASTDENKCVQSDVDSFFKHLIDIGLTTPSSYLSNVLDDNICGADKAPWTTYTTYTTTKNCDIQKKTPKSQSCDTLVVVNVPSPLGNTPHEYKYACECKIPTTEETCDDRKEYMNQWSCGSAQTVRGRSGKDDYELYTYNGVKETKPLGTLKNSKLD
>gi|254952636|gb|ACT97148.1|VAR2CSA[恶性疟原虫]|350aa(SEQ ID NO:30)
KCEKCKSEQSKKNNKNWIWRKFRGTEGGLQEEYANTIGLPPRTQSLCLVVCLDEKGKKTQELKNIRTNSELLKEWIIAAFHEGKNLKPSHQNKNSGNKENLCKALKYSFADYGDLIKGTSIWDNDFTKDLELNLQKIFGKLFRKYIKKNNTAEQHTSYSSLDELRESWWNTNKKYIWTAMKHGAEMNGTTCNADGSVTGSSDSGSTTCSGDNGSISCDDIPTIDLIPQYLRFLQEWVEHFCKQRQEKVNAVINSCNSCKNTSSKTKLGDTCNSDCKTKCKIECEKYKTFIEKCVTAAGGTSGSPWSKRWDQIYKRYSKYIEDAKRNRKAGTKNCGPSSTTSTAESKCVQS
>gi|254952638|gb|ACT97149.1|VAR2CSA[恶性疟原虫]|330aa(SEQ ID NO:31)
KCDKCKSEQSKKNNKNWIWRKYSGNGEGLQKEYANTIGLPPRTHSLYLVCLHEKEGKTQELKNIRTNSELLKEWIIAAFHEGKNLKTTYLENKNDENKKKLCKALKYSFADYGDLIKGTSIWDNDFTKDLELNLQKIFGKLFRKYIKKNIASDENTLYSSLDELRESWWNTNKKYIWTAMKHGAEMNGTTCSSGSGDNGSISCDDIPTIDLIPQYLRFLQEWVGHFCKQRQEKVNAVITNCNSCKESGGTCNSDCEKKCKIECEKYKKFIEECRTAAGGTSGSPWSKRWDQIYKMYSKYIEDAKRNRKAGTKNCGPSSTTSTAE SKCVQS
>gi|254952628|gb|ACT97144.1|VAR2CSA[恶性疟原虫]|334aa(SEQ ID NO:32)
KCDKCKSEQSKKNNKNWIWRKYSGNGEGLQKEYANTIGLPPRTHSLYLVCLHEKEGKTQHKTISTNSELLKEWIIAAFHEGKNLKKRYPQNNNSGNKKKLCKDLKYSFADYGDLIKGTSIWDNEYTKDLELNLQKAFGKLFRKYIKKNIASDENTLYSSLDELRESWWNTNKKYIWLAMKHGAEMNGTMCNADGSVTGSGSSCDDMSTIDLIPQYLRFLQEWVEHFCEQRQAKVKDVINSCKSCKESGDTCNSDCEKKCKNKCDAYKTFIEEFCTADGGTAGSPWSKRWDQIYKRYSKYIEDAKRNRKAGTKNCGTSSGAN SGVTTTENKCVQS
>gi|254952630|gb|ACT97145.1|VAR2CSA[恶性疟原虫]|350aa(SEQ ID NO:33)
KCDKCKSGTSTVNKNWIWKKYSGKEEGLQKEYANTIALPPRTHSLYLVCLHEKGKKTQELKNIRTNSELLKEWIIAAFHEGKNLKTSPQNNNSGNKKKLCKALKYSFADYGDLIKGTSIWDNDFTKDLELNLQKIFGKLFRKYIKKNNTAEQHTSYSSLDELRESWWNTNKKYIWLAMKHGAEMNGTTCCGDGSVTGSSDSGSTTCSGDNGSISCDDMPTTDFIPQYLRFLQEWVEHFCKQRQEKVKHVMESCKSCKECGDTCNGECKTECEKKCKNKCEAYKTFIEKCVSADGGTSGSSWSKRWDQIYMRYSKYIEDAKRNRKAGTKNCGTSSTTNAAASTAENKCVQS
>P13 745个氨基酸|647aa(SEQ ID NO:34)
DYIKDDPYSAEYATKLSFILNPSDANTSSGETANHNDEVCNCNESEIASVELAPISDSSSNKTCITHSFIGANKKKECKDVKLGVREKDKDLKICVIEDDSLRGVENCCCQDLLGILQENCSDNKSGSSSNGSCDKNSEDECQKKLENVFASLKNGYKCDKCKSGTSTVNKKWIWRKYSGNGEGLQKEYANTIGLPPRTHSLYLVCLHEKEGKTQHKTISTNSELLKEWIIAAFHEGKNLKTSHQNNNSGNKKKLCKALKYSFADYGDLIKGTSIWDNDFTKDLELNLQKIFGKLFRKYIKKNIASDENTSYSSLDELRESWWNTNKKYIWLAMKHGAEMNSTMCNGDGSVTGSSDSGSTTCSGDNGSISCDDIPTIDLIPQYLRFLQEWVEHFCKQRQEKVKDVITNCKSCKESGDTCNSDCEKKCKNKCEAYKKFIEERRTAAQGTAESSWVKRWDQIYMRYSKYIEDAKRNRKAGTKSCGPSSTTNAAASTAENKCVQSDIDSFFKHLIDIGLTTPSSYLSIVLDDNICGADNAPWTTYTTYTTTKNCDIKKKTPKPQSCDTLVVVNVPSPLGNTPHEYKYACQCRTPNKQESCDDRKEYMNQWSSGSAQTVRGRSTNNDYELYTYNGVKETKPLGTLKNSKLD
>gi|254952608|gb|ACT97134.1|VAR2CSA[恶性疟原虫]|341aa(SEQ ID NO:35)
KCDKCKSGTSTVNKKWIWRKSSGNKEGLQKEYANTIGLPPRTQSLYLGNLPKLENVCEDVKDINFDTKEKFLAGCLIVSFHEGKNLKTSHEKKNDDNGKKLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQKIFGKLFRKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWTAMKHGAGMNITTCCGDGSSGENQTNSCDDIPTIDLIPQYLRFLQEWVEHFCKQRQEKVNAVVTNCKSCKESGGTCNGECKTKCKNKCEVYKTFIDNVGDGTAGSPWVKRWDQIYKRYSKHIEDAKRNRKAGTKNCGITTGTISG ESSGATSGVTTTENKCVQS
>7g8 745个氨基酸|632aa(SEQ ID NO:36)
NYIKDDPYSKEYVTKLSFIPNSSDANTSSEKIQKNNDEVCNPNESGISSVEQAQTSGPSSNKTCITHSSIKANKKKECKDVKLGVRENDKDLKICVIEDTSLSGVDNCCCQDLLGILQENCSDNKRGSSSNDSCDNKNQDECQKKLDEALESLHNGYKNQKCKSGTSTVNKKWIWKKSSGNKEGLQKEYANTIGLPPRTQSLYLGNLPKLENVSKGVTDIIYDTKEKFLAGCLIVSFHEGKNLKTSHEKKNDDNGKKLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQKAFGKLFRKYIKKNISAEQDTSYSSLDELRESWWNTNKKYIWIAMKHGAGMNGTTCCGDGSSGENQTNSCDDIPTIDLIPQYLRFLQEWVEHFCEQRQAKVKDVITNCKSCKNTSGERKIGGTCNGECKTKCKNKCEAYKTFIEHCKGGDGTAGSSWVKRWDQIYKRYSKHIEDAKRNRKAGTKSCGTSTAENKCVQSDIDSFFKHLIDIGLTTPSSYLSIVLDENNCGEDKAPWTTYTTTKNCDIQKDKSKSQSSDTLVVVNVPSPLGNTPHGYKYACQCKIPTTEETCDDRKEYMNQWSCGSARTMKRGYKNDNYELCKYNGVDVKPTTVRSSSTKLD
>Indo 745个氨基酸|639aa(SEQ ID NO:37)
DYIKGDPYSAEYVTKLSFIPNSSDANNPSEKIQKNNDEVCNCNESEISSVGQASISDPSSNKTCNTHSSIKANKKKVCKDVKLGVRENDKVLKICVIEHTSLRGVDNCCFKDLLGILQEPRIDKNQSGSSSNGSCDKNSEEACEKNLEKVLASLTNGYKCDKCKSGTSRSKKKWIWKKYSGKEGGLQEEYANTIGLPPRTQSLCLVVCLDEKEGKTQELKNISTNSELLKEWIIAAFPEGKNLKPSPEKKKGDNGKKLCKDLKYSFADYGDLIKGTSIWDNEYTKDLELNLQKIFGKLFRKYIKKNIASDENTLYSSLDELRESWWNTNKKYIWLAMKHGAGMNSTMCNADGSVTGSGSSCDDMPTIDLIPQYLRFLQEWVEHFCKQRQEKVKPVIENCNSCKNTSSERKIGGTCNSDCKTECKNKCEVYKKFIEDCKGGDGTAGSSWSKRWDQIYKRYSKYIEDAKRNRKAGTKNCGPSSTTNAAENKCVQSDIDSFFKHLIDIGLTTPSSYLSTVLDDNICGEDNAPWTTYTTYTTTKNCDKDKKKSKSQSCDTLVVVNVPSPLGNTPHEYKYACECRTPNKQESCDDRKEYMNQWISDNTKNPKGSGSGKDYYELYTYNGVDVKPTTVRSSSTKLD
>MC 745个氨基酸|655aa(SEQ ID NO:38)
DYIKGDPYFAEYATKLSFILNSSDANTSSGETANHNDEACNCNESEISSVEHASISDPSSNKTCNTHSSIKANKKKVCKHVKLGVRENDKDLRVCVIEHTSLSGVENCCFKDFLRILQENCSDNKSGSSSNGSCDKNNEEACEKNLEKVFASLTNCYKCEKCKSEQSKKNNKKWTWRKSSGNKGGLQEEYANTIGLPPRTQSLCLVVCLDEKEGKKTQELKNIRTNSELLKEWIIAAFHEGKNLKPSHEKKNDDNGKKNDDNNSKLCKDLKYSFADYGDLIKGTSIWDNEYTKDLELNLQKIFGKLFRKYIKKNIASDENTLYSSLDELRESWWNTNKKYIWLAMKHGAEMNGTTCNADGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVEHFCKQRQAKVKDVIENCKSCKESGNKCKTECKNKCEAYKKFIENCKGGDGTAGSSWVKRWDQIYMRYSKYIEDAKRNRKAGTKNCGPSSITNVSASTDENKCVQSDIDSFFKHLIDIGLTTPSSYLSIVLDDNICGDDKAPWTTYTTYTTYTTYTTYTTYTTYTTTKNCDKERDKSKSQSCNTAVVVNVPSPLGNTPHEYKYACECRTPSNKELCDDRKEYMNQWSSGSAQTVRDRSGKDYYELYTYNGVKETKLPKKLNS SKLD
>gi|254952650|gb|ACT97155.1|VAR2CSA[恶性疟原虫]|347aa(SEQ ID NO:39)
KCDKCKSEQSKKNNKYWIWKKSSVKEEGLQKEYANTIALPPRTHSLCLVVCLDEKGKKTQELKNISTNSELLKERIIAAFHEGKNLKTTYLEKKNADNNSKLCKALKYSFADYGDLIKGTSIWDNEYTKDLELNLQQIFGKLFRKYIKKNNTAEQHTLYSSLDELRESWWNTNKKYIWLAMKHGAGMNGTTCCGDGSVTGSSDSGSTTCSGDNGSISCDDMPTTDFIPQYLRFLQEWVEHFCKQRQEKVKDVIENCNSCKNNLGKTEINEKCKTECKNKCEAYKNFIEKFCTADGGTSGSPWSKRWDQIYKRYSKYIEDAKRNRKAGTKNCGTSSTTSTAENKCVQS
>gi|254952648|gb|ACT97154.1|VAR2CSA[恶性疟原虫]|335aa(SEQ ID NO:40)
KCEKCKSGTSTVNKYWIWRKSSGNKEGLQKEYANTIALPPRTHSLCLVVCLDEKEGKTQELKNISTNSELLKERIIAAFHEGENLKTSHEKKKGDDGKKNADNNSKLCKALKYSFADYGDLIKGTSIWDNEYTKDLELNLQKIFGKLFRKYIKKNIASDENTSYSSLDELRESWWNTNKKYIWLAMKHGAGMNGTTCSCSGDSSDDMPTTDFIPQYLRFLQEWVEHFCKQRQENVNAVIENCNSCKECGGTCNSDCEKKCKTECKNKCEAYKNFIEKFCTADGGTSGYSWSKRWDQIYKRYSKYIEDAKRNRKAGTKSCGTSS TTSTAESKCVQS
>ghana2 745个氨基酸|667aa(SEQ ID NO:41)
SYVKNNPYSKEYVTKLSFILNPSDANNPSETPSKYYDEVCNCNESGIACVGQAQTSGPSSNKTCITHSFIGANKKKVCKDVKLGVREKDKDLKICVIEDTYLSGVDNCCFKDFLGMLQENCSDNKSGSSSNGSCNNKNQDECEKNLDEALASLTNGYKCEKCKSGTSTVNKYWIWRKSSGNKEGLQKEYANTIALPPRTHSLCLVVCLDEKEGKTQHKTISTNSELLKEWIIAAFHEGKNLKTSHEKKKGDDGKKNADNNSKLCKALKYSFADYGDLIKGTSIWDNDFTKDLELNLQKIFGKLFRKYIKKNIASDENTSYSSLDELRESWWNTNKKYIWLAMKHGAGMNSTTCCGDGSVTGSSDSGSTTCCGDGSVTGSGSSCDDMPTTDFIPQYLRFLQEWVEHFCKQRQENVNAVIENCNSCKECGGTCNSDCEKKCKTECKGECDAYKEFIEKCNGGAAEGTSGSSWSKRWDQIYKRYSKYIEDAKRNRKAGTKNCGTSSTTSTAESKCVQSDIDSFFKHLIDIGLTTPSSYLSIVLDENICGADNAPWTTYTTYTTYTTYTTTEKCNKETDKSKLQQCNTSVVVNVPSPLGNTPHGYKYVCECRTPNKQETCDDRKEYMNQWISDNTKNPKGSRSTNNDYELYTY NGVQIKPTTVRSNSTKLD
>gi|254952634|gb|ACT97147.1|VAR2CSA[恶性疟原虫]|348aa(SEQ ID NO:42)
KCDKCKSEQSKKNNKNWIWKKSSGNEKGLQKEYANTIGLPPRTQSLCLVVCLDEKEGKTQELKNIRTNSELLKEWIIAAFHEGKNLKTSHEKKKGDNNSKLCKDLKYSFADYGDLIKGTSIWDNEYTKDLELNLQNNFGKLFRKYIKKNIASDENTSYSSLDELRESWWNTNKKYIWLAMKHGAGMNSTTCSSGSGSTTCSSGSGSTTCSSGSGDSCDDMPTIDLIPQYLRFLQEWVEHFCKQRQEKVNAVIKNCNSCKESGGTCNGECKTECKNKCEAYKTFIEEFCTADGGTSGSPWSKRWDQIYKMYSKHIEDAKRNRKAGTKNCGPSSTTNVSVSTDENKCVQS
>ghana1 745个氨基酸|652aa(SEQ ID NO:43)
DYIKDDPYFAEYVTKLSFILNSSDANNPSGETANHNDEVCNPNESGIASVEQAQTSDPSSNKTCNTHSSIKANKKKVCKHVKLGVRENDKDLKICVIEHTSLSGVENCCCQDFLRILQENCSDNKSGSSSNGSCNNKNQEACEKNLEKVLASLTNCYKCDKCKSEQSKKNNKNWIWKKSSGNEKGLQKEYANTIGLPPRTQSLCLVVCLDEKEGKTQELKNIRTNSELLKEWIIAAFHEGKNLKKRYPQNKNDDNNSKLCKDLKYSFADYGDLIKGTSIWDNEYTKDLELNLQNNFGKLFRKYIKKNISTEQDTLYSSLDELRESWWNTNKKYIWLAMKHGAGMNSTTCSSGSGSTTCSSGSGSTTCSSGSGDSCDDMPTTDFIPQYLRFLQEWVEHFCKQRQEKVNAVIKNCNSCKESGGTCNGECKTECKNKCEAYKTFIEEFCTADGGTSGSPWSKRWDQIYKMYSKHIEDAKRNRKAGTKNCGPSSTTNVSVSTDENKCVQSDIDSFFKHLIDIGLTTPSSYLSIVLDDNICGEDKAPWTTYTTYTTTKKCNKETDKSKSQSCNTAVVVNVPSPLGNTPHGYKYACECKIPTTEETCDDRKEYMNQWIIDTSKKQKGSGSGKDDYELYTYNGVDVKPTTVRSNSTK LD
>V1S1 745个氨基酸|628aa(SEQ ID NO:44)
DYIKDDPYSAQYTTKLSFILNPSDANTSSEKIQKNNDEACNCNESGISSVGQAQTSGPSSNKTCITHSSIKANKKKVCKDVKLGINNNDKVLRVCVIEDTSLSGVDNCCCQDLLGILQENCSDNKRGSSSNGSCNNNNEEACEKNLDEAPASLHNGYKNQKCKSGTSRSKKKWIWKKSSGNEKGLQEEYANTIGLPPRTQSLCLVCLHEKEGKTQHKTISTNSELLKEWIIAAFHEGKNLKTSHEKKNDDNGKKLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQKAFGKLFRKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWIAMKHGAGMNGTTCSCSGDSSNDMPTIDLIPQYLRFLQEWVEHFCEQRQAKVKDVITNCKSCKESGNKCKTECKTKCKDECEKYKTFIEDCNGGGTGTAGSSWVKRWDQIYKRYSKHIEDAKRNRKAGTKNCGPSSITNAAASTDENKCVQSDIDSFFKHLIDIGLTTPSSYLSNVLDENSCGDDKAPWTTYTTYTTTKNCDIQKDKSKSQPINTSVVVNVPSPLGNTPYRYKYACECKIPTTEESCDDRKEYMNQWSCGSARTMKRGYKNDNYELCKYNGVDVKPTTVRSNSSKLD
>raj116_var25 745个氨基酸|653aa(SEQ ID NO:45)
DYIKGDPYFAEYATKLSFILNPSDTENASETPSKYYDEACNPNESEIASVEQAQTSGPSSNKTCITHSSIKTNKKKECKDVKLGVRENDKDLKICVIEDTSLSGVDNCCFKDLLGILQENCSDNKRGSSSNDSCNNNNEEACEKNLDEALASLTNGYKCDKCKSGTSTVNKKWTWRKSSGNEEGLQKEYANTIGLPPRTQSLCLVCLHEKEGKTKHKTISTNSELLKEWIIAAFHEGKNLKTSHEKKNDDNGKKLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQKAFGKLFRKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWTAMKHGAEMNGTTCSSGSGDNGDSSITGSSDSGSTTCSGDNGSISCDDIPTTDFIPQYLRFLQEWVEHFCEQRQAKVKDVINSCNSCNESGGTCNGECKTKCKDECEKYKKFIEDCNGGDGTAGSSWVKRWDQIYKRYSKHIEDAKRNRKAGTKNCGPSSITNAAASTDENKCVQSDVDSFFKHLIDIGLTTPSSYLSIVLDENSCGDDKAPWTTYTTYTTTEKCNKERDKSKSQSSDTLVVVNVPSPLGNTPHEYKYACECKIPTNEETCDDRKDYMNQWISDTSKKQKGSGSGKDYYELYTYNGVQIKQAAGRSSSTKL D
>gi|31323048|gb|AAP37940.1|var2csa[恶性疟原虫]|490aa(SEQ ID NO:46)
KCDKCKSEQSKKNNNKWIWKKYSGNGEGLQKEYANTIGLPPRTQSLCLVCLHEKEGKTQHKTISTNSELLKEWIIAAFHEGKNLKKRYPQNKNDDNNSKLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQKAFGKLFRKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWTAMKHGAEMNGTTCSSGSGDNGDSSCDDIPTIDLIPQYLRFLQEWVEHFCKQRQAKVKDVINSCNSCKNTSGERKIGGTCNSDCEKKCKVACDAYKTFIEECRTAVGGTAGSSWVKRWDQIYKRYSKHIEDAKRNRKAGTKNCGPSSTTNAAENKCVQSDIDSFFKHLIDIGLTTPSSYLSNVLDENSCGADKAPWTTYTTYTTYTTYTTYTTTEKCNKERDKSKSQQSNTSVVVNVPSPLGNTPHEYKYACECKIPTTEETCDDRKEYMNQWIIDNTKNPKGSGSTDNDYELYTYNGVQIKQAAGRSS STKLD
>gi|254952620|gb|ACT97140.1|VAR2CSA[恶性疟原虫]|335aa(SEQ ID NO:47)
KCEKCKSGTSTVNNKWIWRKSSGKEGGLQKEYANTIGLPPRTQSLYLGNLPKLENVCKGVTDIIYDTKEKFLSGCLIAAFHEGKNLKTTYLEKKNDDNGKKLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQKIFGKLFRKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWIAMKHGAGMNGTTCSSGSGDSSNDIPTTDFIPQYLRFLQEWVENFCEQRQAKVKPVIENCNSCKESGGTCNGECKTKCKVACDAYKKFIDGTGSGGGSRPTGIAGSSWSKRWDQIYKRYSKHIEDAKRNRKAGTKNCGPSSITNVS VSTDENKCVQS
>T2C6 745个氨基酸|637aa(SEQ ID NO:48)
NYIKDDPYSKEYVTKLSFIPNSSDANTSSEKIQKNNDEVCNPNESGISSVEQAQTSDPSSNKTCITHSSIKANKKKECKDVKLGVRENDKDLKICVIEHTSLSGVDNCCFKDFLRMLQEPRIDKNQRGSSSNGSCDKNSEEACEKNLDEALASLTNGYKCDKCKSEQSKKNNNKWIWKKFPGKEGGLQEEYANTIGLPPRTQYLCLVVCLDEKEGKTQELKNIRTNSELLKEWIIAAFHEGKNLKTTYPQKKNDDNGKKLCKDLKYSFADYGDLIKGTSIWDNEYTKNVELNLQNNFGKLFRKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWLAMKHGAEMNSTTCCGDGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVEHFCKQRQAKVKDVITNCNSCKESGNKCKTECKNKCKDECEKYKKFIEACGTAVGGTGTAGSPWSKRWDQIYKRYSKHIEDAKRNRKAGTKNCGPSSTTNAAENKCVQSDIDSFFKHLIDIGLTTPSSYLSIVLDDNICGADKAPWTTYTTYTTENCDIQKKTPKSQSCDTLVVVNVPSPLGNTPHGYKYACQCRTPNKQESCDDRKEYMNQWIIDNTKNPKGSGSGKDYYELCKYNGVKETKPLGTLKNSKLD
>gi|254952632|gb|ACT97146.1|VAR2CSA[恶性疟原虫]|330aa(SEQ ID NO:49)
KCDKCKSEQSKKNNNKWIWRKFPGKEGGLQKEYANTIGLPPRTQSLCLVCLHEKEGKTQHKTISTNSELLKEWIIAAFHEGKNLKTTYLEKKNAENKKKLCKALKYSFADYGDLIKGTSIWDNEYTKDLELNLQKIFGKLFRKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWTAMKHGAGMNGTMCNADGSVTGSGSSCDDMPTTDFIPQYLRFLQEWVEHFCKQRQAKVKDVIENCKSCKESGNKCKTECKNKCDAYKTFIEECGTAVGGTAGSSWVKRWDQIYKRYSKHIEDAKRNRKAGTKNCGTSSTTNAAA STAENKCVQS
>gi|90193487|gb|ABD92339.1|红细胞膜蛋白1[恶性疟原虫]|269aa(SEQ ID NO:50)
NYIKDDPYSKEYVTKLSFILNSSDAENASETPSKYYDEACNCNESGISSVEQASISDRSSQKACNTHSFIGANKKKVCKHVKLGVRENDKDLKICVIEDDSLRGVENCCFKDFLRMLQEPRIDKNQRGSSSNDSCNNNNEEACEKNLDEALASLHNGYKNQKCKSEQSKKNNNKWIWKKSSGKEGGLQKEYANTIGLPPRTQSLCLVCLHEKEGKTQHKTISTNSELLKEWIIDAFHEGKNLKTTYLEKKKGDNGKKLCKALKYSFADY
>gi|254952646|gb|ACT97153.1|VAR2CSA[恶性疟原虫]|347aa(SEQ ID NO:51)
KCDKCKSEQSKKNNKNWIWKKSSGKEGGLQKEYANTIALPPRTQSLCLVVCLHEKEGKTQHKTISTNSELLKEWIIDAFHEGKNLKTTYLEKQNADNGKKNADNNSKLCKDLKYSFADYGDLIKGTSIWDNEYTKDLELNLQQIFGKLFRKYIKKNIASDENTLYSSLDELRESWWNTNKKYIWTAMKHGAEMNGTTCSSGSGDSSSGENQTNSCDDIPTIDLIPQYLRFLQEWVEHFCEQRQAKVKDVITNCKSCKESGGTCNSDCKTKCKGECEKYKKFIEKCKGGGTEGTSGSSWVKRWYQIYMRYSKYIEDAKRNRKAGTKSCGTSSGANSGVTTTESKCVQS
>gi|90193485|gb|ABD92338.1|红细胞膜蛋白1[恶性疟原虫]|269aa(SEQ ID NO:52)
DYIKDDPYSKEYTTKLSFILNSSDANTSSEKIQKNNDEVCNPNESEISSVEQAQTSRPSSNKTCITHSSIKANKKKVCKDVKLGVRENDKVLRVCVIEHTSLSGVENCCCQDLLGILQENCSDNKRGSSSNGSCDKNSEEACEKNLDEALASLTNCYKNQKCKSEQSKKNNNKWIWKKSSGNEKGLQKEYANTIGLPPRTQSLCLVCLHEKEGKTQELKNISTNSELLKEWIIAAFHEGKNLKTTYPQNKNDDNGKKLFKDLKYSFADY
>MTS1 745个氨基酸|646aa(SEQ ID NO:53)
DYIKDDPYSKEYTTKLSFILNSSDANTSSEKIQKNNDEVCNPNESEISSVEQAQTSRPSSNKTCITHSSIKANKKKVCKDVKLGVRENDKVLRVCVIEHTSLSGVENCCCQDLLGILQENCSDNKRGSSSNGSCDKNSEEACEKNLDEALASLTNCYKNQKCKSEQSKKNNNKWIWKKSSGKEGGLQKEYANTIGLPPRTQSLYLGNLPKLENVCKGVTDINFDTKEKFLAGCLIAAFHEGKNLKTTYLEKKNDDNGKKLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQKAFGKLFRKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWTAMKHGAGMNGTTCSSGSGDSSNDIPTTDFIPQYLRFLQEWVENFCEQRQAKVKDVIENCNSCKNTSGERKIGDTCNSDCEKKCKDECEKYKKFIEDCKGGDGTAGSSWVKRWDQIYKRYSKHIEDAKRNRKAGTKNCGITTGTISGESSGATSGVTTTENKCVQSDIDSFFKHLIDIGLTTPSSYLSNVLDDNICGEDNAPWTTYTTYTTEKCNKETDKSKSQQSNTAVVVNVPSPLGNTPHGYKYACECKIPTTEETCDDRKEYMNQWSCGSAQTVRDRSGKDDYELCKYNGVQIKQAAGTLKNSKLD
>Q8I639(Q8I639_PLAF7)恶性疟原虫(分离株3D7),632aa胞外部分(SEQ ID NO:54)
NYIKGDPYFAEYATKLSFILNSSDANNPSEKIQKNNDEVCNCNESGIASVEQEQISDPSSNKTCITHSSIKANKKKVCKHVKLGVRENDKDLRVCVIEHTSLSGVENCCCQDFLRILQENCSDNKSGSSSNGSCNNKNQEACEKNLEKVLASLTNCYKCDKCKSEQSKKNNKNWIWKKSSGKEGGLQKEYANTIGLPPRTQSLCLVVCLDEKGKKTQELKNIRTNSELLKEWIIAAFHEGKNLKPSHEKKNDDNGKKLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQKIFGKLFRKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWLAMKHGAGMNSTTCCGDGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVEHFCKQRQEKVKPVIENCKSCKESGGTCNGECKTECKNKCEVYKKFIEDCKGGDGTAGSSWVKRWDQIYKRYSKYIEDAKRNRKAGTKNCGPSSTTNAAENKCVQSDIDSFFKHLIDIGLTTPSSYLSIVLDDNICGADKAPWTTYTTYTTTEKCNKETDKSKLQQCNTAVVVNVPSPLGNTPHGYKYACQCKIPTNEETCDDRKEYMNQWSCGSARTMKRGYKNDNYELCKYNGVDVKPTTVRSNSSKLD
>Q8I639(Q8I639_PLAF7)恶性疟原虫(分离株3D7),完整的2730aa胞外部分 (SEQID NO:55)
MDKSSIANKIEAYLGAKSDDSKIDQSLKADPSEVQYYGSGGDGYYLRKNICKITVNHSDSGTNDPCDRIPPPYGDNDQWKCAIILSKVSEKPENVFVPPRRQRMCINNLEKLNVDKIRDKHAFLADVLLTARNEGERIVQNHPDTNSSNVCNALERSFADIADIIRGTDLWKGTNSNLEQNLKQMFAKIRENDKVLQDKYPKDQNYRKLREDWWNANRQKVWEVITCGARSNDLLIKRGWRTSGKSNGDNKLELCRKCGHYEEKVPTKLDYVPQFLRWLTEWIEDFYREKQNLIDDMERHREECTSEDHKSKEGTSYCSTCKDKCKKYCECVKKWKSEWENQKNKYTELYQQNKNETSQKNTSRYDDYVKDFFKKLEANYSSLENYIKGDPYFAEYATKLSFILNSSDANNPSEKIQKNNDEVCNCNESGIASVEQEQISDPSSNKTCITHSSIKANKKKVCKHVKLGVRENDKDLRVCVIEHTSLSGVENCCCQDFLRILQENCSDNKSGSSSNGSCNNKNQEACEKNLEKVLASLTNCYKCDKCKSEQSKKNNKNWIWKKSSGKEGGLQKEYANTIGLPPRTQSLCLVVCLDEKGKKTQELKNIRTNSELLKEWIIAAFHEGKNLKPSHEKKNDDNGKKLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQKIFGKLFRKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWLAMKHGAGMNSTTCCGDGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVEHFCKQRQEKVKPVIENCKSCKESGGTCNGECKTECKNKCEVYKKFIEDCKGGDGTAGSSWVKRWDQIYKRYSKYIEDAKRNRKAGTKNCGPSSTTNAAENKCVQSDIDSFFKHLIDIGLTTPSSYLSIVLDDNICGADKAPWTTYTTYTTTEKCNKETDKSKLQQCNTAVVVNVPSPLGNTPHGYKYACQCKIPTNEETCDDRKEYMNQWSCGSARTMKRGYKNDNYELCKYNGVDVKPTTVRSNSSKLDDKDVTFFNLFEQWNKEIQYQIEQYMTNTKISCNNEKNVLSRVSDEAAQPKFSDNERDRNSITHEDKNCKEKCKCYSLWIEKINDQWDKQKDNYNKFQRKQIYDANKGSQNKKVVSLSNFLFFSCWEEYIQKYFNGDWSKIKNIGSDTFEFLIKKCGNDSGDGETIFSEKLNNAEKKCKENESTNNKMKSSETSCDCSEPIYIRGCQPKIYDGKIFPGKGGEKQWICKDTIIHGDTNGACIPPRTQNLCVGELWDKRYGGRSNIKNDTKESLKQKIKNAIQKETELLYEYHDKGTAIISRNPMKGQKEKEEKNNDSNGLPKGFCHAVQRSFIDYKNMILGTSVNIYEYIGKLQEDIKKIIEKGTTKQNGKTVGSGAENVNAWWKGIEGEMWDAVRCAITKINKKQKKNGTFSIDECGIFPPTGNDEDQSVSWFKEWSEQFCIERLQYEKNIRDACTNNGQGDKIQGDCKRKCEEYKKYISEKKQEWDKQKTKYENKYVGKSASDLLKENYPECISANFDFIFNDNIEYKTYYPYGDYSSICSCEQVKYYEYNNAEKKNNKSLCHEKGNDRTWSKKYIKKLENGRTLEGVYVPPRRQQLCLYELFPIIIKNKNDITNAKKELLETLQIVAEREAYYLWKQYHAHNDTTYLAHKKACCAIRGSFYDLEDIIKGNDLVHDEYTKYIDSKLNEIFDSSNKNDIETKRARTDWWENEAIAVPNITGANKSDPKTIRQLVWDAMQSGVRKAIDEEKEKKKPNENFPPCMGVQHIGIAKPQFIRWLEEWTNEFCEKYTKYFEDMKSNCNLRKGADDCDDNSNIECKKACANYTNWLNPKRIEWNGMSNYYNKIYRKSNKESEDGKDYSMIMEPTVIDYLNKRCNGEINGNYICCSCKNIGENSTSGTVNKKLQKKETQCEDNKGPLDLMNKVLNKMDPKYSEHKMKCTEVYLEHVEEQLKEIDNAIKDYKLYPLDRCFDDKSKMKVCDLIGDAIGCKHKTKLDELDEWNDVDMRDPYNKYKGVLIPPRRRQLCFSRIVRGPANLRNLKEFKEEILKGAQSEGKFLGNYYNEDKDKEKALEAMKNSFYDYEYIIKGSDMLTNIQFKDIKRKLDRLLEKETNNTEKVDDWWETNKKSIWNAMLCGYKKSGNKIIDPSWCTIPTTETPPQFLRWIKEWGTNVCIQKEEHKEYVKSKCSNVTNLGAQESESKNCTSEIKKYQEWSRKRSIQWEAISEGYKKYKGMDEFKNTFKNIKEPDANEPNANEYLKKHCSKCPCGFNDMQEITKYTNIGNEAFKQIKEQVDIPAELEDVIYRLKHHEYDKGNDYICNKYKNINVNMKKNNDDTWTDLVKNSSDINKGVLLPPRRKNLFLKIDESDICKYKRDPKLFKDFIYSSAISEVERLKKVYGEAKTKVVHAMKYSFADIGSIIKGDDMMENNSSDKIGKILGDGVGQNEKRKKWWDMNKYHIWESMLCGYKHAYGNISENDRKMLDIPNNDDEHQFLRWFQEWTENFCTKRNELYENMVTACNSAKCNTSNGSVDKKECTEACKNYSNFILIKKKEYQSLNSQYDMNYKETKAEKKESPEYFKDKCNGECSCLSEYFKDETRWKNPYETLDDTEVKNNCMCKPPPPASNNTSDILQKTIPFGIALALGSIAFLFMKKKPKTPVDLLRVLDIPKGDYGIPTPKSSNRYIPYASDRYKGKTYIYMEGDTSGDDDKYIWDL
>FCR3(SEQ ID NO:56)完整的2734aa胞外部分(577aa高亮的对应ID1-DBL2b)
MDSTSTIANKIEEYLGAKSDDSKIDELLKADPSEVEYYRSGGDGDYLKNNICKITVNHSDSGKYDPCEKKLPPYDDNDQWKCQQNSSDGSGKPENICVPPRRERLCTYNLENLKFDKIRDNNAFLADVLLTARNEGEKIVQNHPDTNSSNVCNALERSFADLADIIRGTDQWKGTNSNLEKNLKQMFAKIRENDKVLQDKYPKDQKYTKLREAWWNANRQKVWEVITCGARSNDLLIKRGWRTSGKSDRKKNFELCRKCGHYEKEVPTKLDYVPQFLRWLTEWIEDFYREKQNLIDDMERHREECTREDHKSKEGTSYCSTCKDKCKKYCECVKKWKTEWENQENKYKDLYEQNKNKTSQKNTSRYDDYVKDFFEKLEANYSSLENYIKGDPYFAEYATKLSFILNPSDANNPSGETANHNDEACNCNESGISSVGQAQTSGPSSNKTCITHSSIKTNKKKECKDVKLGVRENDKDLKICVIEDTSLSGVDNCCCQDLLGILQENCSDNKRGSSSNDSCDNKNQDECQKKLEKVFASLTNGYKCDKCKSGTSRSKKKWIWKKSSGNEEGLQEEYANTIGLPPRTQSLYLGNLPKLENVCEDVKDINFDTKEKFLAGCLIVSFHEGKNLKKRYPQNKNSGNKENLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQNNFGKLFGKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWTAMKHGAEMNITTCNADGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVENFCEQRQAKVKDVITNCKSCKESGNKCKTECKTKCKDECEKYKKFIEACGTAGGGIGTAGSPWSKRWDQIYKRYSKHIEDAKRNRKAGTKNCGTSSTTNAAASTDENKCVQSDIDSFFKHLIDIGLTTPSSYLSNVLDDNICGADKAPWTTYTTYTTTEKCNKERDKSKSQSSDTLVVVNVPSPLGNTPYRYKYACQCKIPTNEETCDDRKEYMNQWSCGSARTMKRGYKNDNYELCKYNGVDVKPTTVRSNSSKLDGNDVTFFNLFEQWNKEIQYQIEQYMTNANISCIDEKEVLDSVSDEGTPKVRGGYEDGRNNNTDQGTNCKEKCKCYKLWIEKINDQWGKQKDNYNKFRSKQIYDANKGSQNKKVVSLSNFLFFSCWEEYIQKYFNGDWSKIKNIGSDTFEFLIKKCGNNSAHGEEIFNEKLKNAEKKCKENESTDTNINKSETSCDLNATNYIRGCQSKTYDGKIFPGKGGEKQWICKDTIIHGDTNGACIPPRTQNLCVGELWDKSYGGRSNIKNDTKELLKEKIKNAIHKETELLYEYHDTGTAIISKNDKKGQKGKNDPNGLPKGFCHAVQRSFIDYKNMILGTSVNIYEHIGKLQEDIKKIIEKGTPQQKDKIGGVGSSTENVNAWWKGIEREMWDAVRCAITKINKKNNNSIFNGDECGVSPPTGNDEDQSVSWFKEWGEQFCIERLRYEQNIREACTINGKNEKKCINSKSGQGDKIQGACKRKCEKYKKYISEKKQEWDKQKTKYENKYVGKSASDLLKENYPECISANFDFIFNDNIEYKTYYPYGDYSSICSCEQVKYYKYNNAEKKNNKSLCYEKDNDMTWSKKYIKKLENGRSLEGVYVPPRRQQLCLYELFPIIIKNEEGMEKAKEELLETLQIVAEREAYYLWKQYNPTGKGIDDANKKACCAIRGSFYDLEDIIKGNDLVHDEYTKYIDSKLNEIFGSSDTNDIDTKRARTDWWENETITNGTDRKTIRQLVWDAMQSGVRYAVEEKNENFPLCMGVEHIGIAKPQFIRWLEEWTNEFCEKYTKYFEDMKSKCDPPKRADTCGDNSNIECKKACANYTNWLNPKRIEWNGMSNYYNKIYRKSNKESEGGKDYSMIMAPTVIDYLNKRCHGEINGNYICCSCKNIGAYNTTSGTVNKKLQKKETECEEEKGPLDLMNEVLNKMDKKYSAHKMKCTEVYLEHVEEQLNEIDNAIKDYKLYPLDRCFDDQTKMKVCDLIADAIGCKDKTKLDELDEWNDMDLRGTYNKHKGVLIPPRRRQLCFSRIVRGPANLRSLNEFKEEILKGAQSEGKFLGNYYKEHKDKEKALEAMKNSFYDYEDIIKGTDMLTNIEFKDIKIKLDRLLEKETNNTKKAEDWWKTNKKSIWNAMLCGYKKSGNKIIDPSWCTIPTTETPPQFLRWIKEWGTNVCIQKQEHKEYVKSKCSNVTNLGAQASESNNCTSEIKKYQEWSRKRSIRWETISKRYKKYKRMDILKDVKEPDANTYLREHCSKCPCGFNDMEEMNNNEDNEKEAFKQIKEQVKIPAELEDVIYRIKHHEYDKGNDYICNKYKNIHDRMKKNNGNFVTDNFVKKSWEISNGVLIPPRRKNLFLYIDPSKICEYKKDPKLFKDFIYWSAFTEVERLKKAYGGARAKVVHAMKYSFTDIGSIIKGDDMMEKNSSDKIGKILGDTDGQNEKRKKWWDMNKYHIWESMLCGYREAEGDTETNENCRFPDIESVPQFLRWFQEWSENFCDRRQKLYDKLNSECISAECTNGSVDNSKCTHACVNYKNYILTKKTEYEIQTNKYDNEFKNKNSNDKDAPDYLKEKCNDNKCECLNKHIDDKNKTWKNPYETLEDTFKSKCDCPKPLPSPIKPDDLPPQADEPFDPTILQTTIPFGIALALGSIAFLFMKVIYIYIYVCCICMYVCMYVCMYVCMYVCMYVCMHVCMLCVYVIYVFKICIYIEKEKRKK
>BPTI,蛋白酶抑制剂(SEQ ID NO:57)
RPDFCLEPPYTGPCKARIIRYFYNAKAGLCQTFVYGGCRAKRNNFKSAEDCMRT CGGA >PE38,假单胞菌外毒素A(SEQ ID NO：58)(下划线的KDEL表示对根据本发明的构建体可任选的信号序列)
RHRQPRGWEQLEQCGYPVQRLVALYLAARLSWNQVDQVIRNALASPGSGGDLGEAIREQPEQARLALTLAAAESERFVRQGTGNDEAGAANGPADSGDALLERNYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLEERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIRNGALLRVYVPRSSLPGFYRTSLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLETILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYASQPGKPPRKDEL
>PE38LR,PE38的变体(SEQ ID NO:59)(下划线的KDEL表示对根据本发明的构建体可任选的信号序列)
RHRQPRGWEQLYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLEERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIRNGALLRVYVPRSSLPGFYRTSLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLETILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYAS QPGKPPRKDEL
与假单胞菌外毒素A(PE38)的截短片段融合的VAR2CSA多肽的序列融合的VAR2CSA-PE38蛋白可以具有修饰，例如N末端的蛋白酶抑制剂(BPTI)和 /或免疫原性较低的优化的PE38序列(PE38LR)
>BPTI-ID1-ID2aFCR3-PE38LR(SEQ ID NO：60)，下划线的序列对应于FCR3的 ID1结构域，粗体字序列对应于FCR3的DBL2Xb结构域，下划线和粗体序列为ID2aRPDFCLEPPYTGPCKARIIRYFYNAKAGLCQTFVYGGCRAKRNNFKSAEDCMRTCGGANYIKGDPYFAEYATKLSFILNPSDANNPSGETANHNDEACNCNESGISSVGQAQTSGPSSNKTCITHSSIKTN KKKECKDVKLGVRENDKDLKICVIEDTSLSGVDNCCCQDLLGILQENCSDNKRGSSSNDSCDNKNQDECQKKLEKVF ASLTNGYKCD KCKSGTSRSKKKWIWKKSSGNEEGLQEEYANTIGLPPRTQSLYLGNLPKLENVCEDVKDINFDTKEKFLAGCLIVSFHEGKNLKKRYPQNKNSGNKENLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQNNFGKLFGKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWTAMKHGAEMNITTCNADGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVENFCEQRQAKVKDVITNCKSCKESGNKCKTECKTKCKDECEKYKKFIEACGTAGGGIGTAGSPWSKRWDQIYKRYSKHIEDAKRNRKAGTKNCGTSSTTNAAASTDENKCVQSDIDSFFKHLIDIGLTTPSSYLSNVLDDNICGADKAPWTTYTTYTTTEKCNKERDKSKSQSSDTLVVVNVPSPLGNTPYRYKYACQCKIPTNEETCDDRKEYMNQWSCGSARTMKRGYKND NYELCKYNGVDVKPTTVRSNSSKLDRHRQPRGWEQLYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLEERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIRNGALLRVYVPRSSLPGFYRTSLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLETILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYASQP GKPPRKDEL
>BPTI-ID1-ID2aFCR3-PE38(SEQ ID NO:61)
RPDFCLEPPYTGPCKARIIRYFYNAKAGLCQTFVYGGCRAKRNNFKSAEDCMRTCGGANYIKGDPYFAEYATKLSFILNPSDANNPSGETANHNDEACNCNESGISSVGQAQTSGPSSNKTCITHSSIKTNKKKECKDVKLGVRENDKDLKICVIEDTSLSGVDNCCCQDLLGILQENCSDNKRGSSSNDSCDNKNQDECQKKLEKVFASLTNGYKCDKCKSGTSRSKKKWIWKKSSGNEEGLQEEYANTIGLPPRTQSLYLGNLPKLENVCEDVKDINFDTKEKFLAGCLIVSFHEGKNLKKRYPQNKNSGNKENLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQNNFGKLFGKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWTAMKHGAEMNITTCNADGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVENFCEQRQAKVKDVITNCKSCKESGNKCKTECKTKCKDECEKYKKFIEACGTAGGGIGTAGSPWSKRWDQIYKRYSKHIEDAKRNRKAGTKNCGTSSTTNAAASTDENKCVQSDIDSFFKHLIDIGLTTPSSYLSNVLDDNICGADKAPWTTYTTYTTTEKCNKERDKSKSQSSDTLVVVNVPSPLGNTPYRYKYACQCKIPTNEETCDDRKEYMNQWSCGSARTMKRGYKNDNYELCKYNGVDVKPTTVRSNSSKLDPEGGSLAALTAHQACHLPLETFTRHRQPRGWEQLEQCGYPVQRLVALYLAARLSWNQVDQVIRNALASPGSGGDLGEAIREQPEQARLALTLAAAESERFVRQGTGNDEAGAANGPADSGDALLERNYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLEERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIRNGALLRVYVPRSSLPGFYRTSLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLETILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISAL PDYASQPGKPPRKDEL
>ID1-ID2aFCR3-PE38(SEQ ID NO:62)
NYIKGDPYFAEYATKLSFILNPSDANNPSGETANHNDEACNCNESGISSVGQAQTSGPSSNKTCITHSSIKTNKKKECKDVKLGVRENDKDLKICVIEDTSLSGVDNCCCQDLLGILQENCSDNKRGSSSNDSCDNKNQDECQKKLEKVFASLTNGYKCDKCKSGTSRSKKKWIWKKSSGNEEGLQEEYANTIGLPPRTQSLYLGNLPKLENVCEDVKDINFDTKEKFLAGCLIVSFHEGKNLKKRYPQNKNSGNKENLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQNNFGKLFGKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWTAMKHGAEMNITTCNADGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVENFCEQRQAKVKDVITNCKSCKESGNKCKTECKTKCKDECEKYKKFIEACGTAGGGIGTAGSPWSKRWDQIYKRYSKHIEDAKRNRKAGTKNCGTSSTTNAAASTDENKCVQSDIDSFFKHLIDIGLTTPSSYLSNVLDDNICGADKAPWTTYTTYTTTEKCNKERDKSKSQSSDTLVVVNVPSPLGNTPYRYKYACQCKIPTNEETCDDRKEYMNQWSCGSARTMKRGYKNDNYELCKYNGVDVKPTTVRSNSSKLDPEGGSLAALTAHQACHLPLETFTRHRQPRGWEQLEQCGYPVQRLVALYLAARLSWNQVDQVIRNALASPGSGGDLGEAIREQPEQARLALTLAAAESERFVRQGTGNDEAGAANGPADSGDALLERNYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLEERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIRNGALLRVYVPRSSLPGFYRTSLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLETILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYAS QPGKPPRKDEL
>ID1-ID2aFCR3-PE38LR(SEQ ID NO:63)
NYIKGDPYFAEYATKLSFILNPSDANNPSGETANHNDEACNCNESGISSVGQAQTSGPSSNKTCITHSSIKTNKKKECKDVKLGVRENDKDLKICVIEDTSLSGVDNCCCQDLLGILQENCSDNKRGSSSNDSCDNKNQDECQKKLEKVFASLTNGYKCDKCKSGTSRSKKKWIWKKSSGNEEGLQEEYANTIGLPPRTQSLYLGNLPKLENVCEDVKDINFDTKEKFLAGCLIVSFHEGKNLKKRYPQNKNSGNKENLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQNNFGKLFGKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWTAMKHGAEMNITTCNADGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVENFCEQRQAKVKDVITNCKSCKESGNKCKTECKTKCKDECEKYKKFIEACGTAGGGIGTAGSPWSKRWDQIYKRYSKHIEDAKRNRKAGTKNCGTSSTTNAAASTDENKCVQSDIDSFFKHLIDIGLTTPSSYLSNVLDDNICGADKAPWTTYTTYTTTEKCNKERDKSKSQSSDTLVVVNVPSPLGNTPYRYKYACQCKIPTNEETCDDRKEYMNQWSCGSARTMKRGYKNDNYELCKYNGVDVKPTTVRSNSSKLDRHRQPRGWEQLYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLEERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIRNGALLRVYVPRSSLPGFYRTSLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLETILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYASQPGKPPRK DEL
>BPTI-DBL1-ID2aFCR3-PE38LR(SEQ ID NO:64)
RPDFCLEPPYTGPCKARIIRYFYNAKAGLCQTFVYGGCRAKRNNFKSAEDCMRTCGGANHSDSGKYDPCEKKLPPYDDNDQWKCQQNSSDGSGKPENICVPPRRERLCTYNLENLKFDKIRDNNAFLADVLLTARNEGEKIVQNHPDTNSSNVCNALERSFADLADIIRGTDQWKGTNSNLEKNLKQMFAKIRENDKVLQDKYPKDQKYTKLREAWWNANRQKVWEVITCGARSNDLLIKRGWRTSGKSDRKKNFELCRKCGHYEKEVPTKLDYVPQFLRWLTEWIEDFYREKQNLIDDMERHREECTREDHKSKEGTSYCSTCKDKCKKYCECVKKWKTEWENQENKYKDLYEQNKNKTSQKNTSRYDDYVKDFFEKLEANYSSLENYIKGDPYFAEYATKLSFILNPSDANNPSGETANHNDEACNCNESGISSVGQAQTSGPSSNKTCITHSSIKTNKKKECKDVKLGVRENDKDLKICVIEDTSLSGVDNCCCQDLLGILQENCSDNKRGSSSNDSCDNKNQDECQKKLEKVFASLTNGYKCDKCKSGTSRSKKKWIWKKSSGNEEGLQEEYANTIGLPPRTQSLYLGNLPKLENVCEDVKDINFDTKEKFLAGCLIVSFHEGKNLKKRYPQNKNSGNKENLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQNNFGKLFGKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWTAMKHGAEMNITTCNADGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVENFCEQRQAKVKDVITNCKSCKESGNKCKTECKTKCKDECEKYKKFIEACGTAGGGIGTAGSPWSKRWDQIYKRYSKHIEDAKRNRKAGTKNCGTSSTTNAAASTDENKCVQSDIDSFFKHLIDIGLTTPSSYLSNVLDDNICGADKAPWTTYTTYTTTEKCNKERDKSKSQSSDTLVVVNVPSPLGNTPYRYKYACQCKIPTNEETCDDRKEYMNQWSCGSARTMKRGYKNDNYELCKYNGVDVKPTTVRSNSSKLDRHRQPRGWEQLYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLEERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIRNGALLRVYVPRSSLPGFYRTSLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLETILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAIS ALPDYASQPGKPPRKDEL
>BPTI-DBL1-ID2aFCR3-PE38(SEQ ID NO:65)
RPDFCLEPPYTGPCKARIIRYFYNAKAGLCQTFVYGGCRAKRNNFKSAEDCMRTCGGANHSDSGKYDPCEKKLPPYDDNDQWKCQQNSSDGSGKPENICVPPRRERLCTYNLENLKFDKIRDNNAFLADVLLTARNEGEKIVQNHPDTNSSNVCNALERSFADLADIIRGTDQWKGTNSNLEKNLKQMFAKIRENDKVLQDKYPKDQKYTKLREAWWNANRQKVWEVITCGARSNDLLIKRGWRTSGKSDRKKNFELCRKCGHYEKEVPTKLDYVPQFLRWLTEWIEDFYREKQNLIDDMERHREECTREDHKSKEGTSYCSTCKDKCKKYCECVKKWKTEWENQENKYKDLYEQNKNKTSQKNTSRYDDYVKDFFEKLEANYSSLENYIKGDPYFAEYATKLSFILNPSDANNPSGETANHNDEACNCNESGISSVGQAQTSGPSSNKTCITHSSIKTNKKKECKDVKLGVRENDKDLKICVIEDTSLSGVDNCCCQDLLGILQENCSDNKRGSSSNDSCDNKNQDECQKKLEKVFASLTNGYKCDKCKSGTSRSKKKWIWKKSSGNEEGLQEEYANTIGLPPRTQSLYLGNLPKLENVCEDVKDINFDTKEKFLAGCLIVSFHEGKNLKKRYPQNKNSGNKENLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQNNFGKLFGKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWTAMKHGAEMNITTCNADGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVENFCEQRQAKVKDVITNCKSCKESGNKCKTECKTKCKDECEKYKKFIEACGTAGGGIGTAGSPWSKRWDQIYKRYSKHIEDAKRNRKAGTKNCGTSSTTNAAASTDENKCVQSDIDSFFKHLIDIGLTTPSSYLSNVLDDNICGADKAPWTTYTTYTTTEKCNKERDKSKSQSSDTLVVVNVPSPLGNTPYRYKYACQCKIPTNEETCDDRKEYMNQWSCGSARTMKRGYKNDNYELCKYNGVDVKPTTVRSNSSKLDPEGGSLAALTAHQACHLPLETFTRHRQPRGWEQLEQCGYPVQRLVALYLAARLSWNQVDQVIRNALASPGSGGDLGEAIREQPEQARLALTLAAAESERFVRQGTGNDEAGAAnGPADSGDALLERNYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLEERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIRNGALLRVYVPRSSLPGFYRTsLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLETILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYASQPGKPPRKDEL
>DBL1-ID2aFCR3-PE38LR(SEQ ID NO:66)
NHSDSGKYDPCEKKLPPYDDNDQWKCQQNSSDGSGKPENICVPPRRERLCTYNLENLKFDKIRDNNAFLADVLLTARNEGEKIVQNHPDTNSSNVCNALERSFADLADIIRGTDQWKGTNSNLEKNLKQMFAKIRENDKVLQDKYPKDQKYTKLREAWWNANRQKVWEVITCGARSNDLLIKRGWRTSGKSDRKKNFELCRKCGHYEKEVPTKLDYVPQFLRWLTEWIEDFYREKQNLIDDMERHREECTREDHKSKEGTSYCSTCKDKCKKYCECVKKWKTEWENQENKYKDLYEQNKNKTSQKNTSRYDDYVKDFFEKLEANYSSLENYIKGDPYFAEYATKLSFILNPSDANNPSGETANHNDEACNCNESGISSVGQAQTSGPSSNKTCITHSSIKTNKKKECKDVKLGVRENDKDLKICVIEDTSLSGVDNCCCQDLLGILQENCSDNKRGSSSNDSCDNKNQDECQKKLEKVFASLTNGYKCDKCKSGTSRSKKKWIWKKSSGNEEGLQEEYANTIGLPPRTQSLYLGNLPKLENVCEDVKDINFDTKEKFLAGCLIVSFHEGKNLKKRYPQNKNSGNKENLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQNNFGKLFGKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWTAMKHGAEMNITTCNADGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVENFCEQRQAKVKDVITNCKSCKESGNKCKTECKTKCKDECEKYKKFIEACGTAGGGIGTAGSPWSKRWDQIYKRYSKHIEDAKRNRKAGTKNCGTSSTTNAAASTDENKCVQSDIDSFFKHLIDIGLTTPSSYLSNVLDDNICGADKAPWTTYTTYTTTEKCNKERDKSKSQSSDTLVVVNVPSPLGNTPYRYKYACQCKIPTNEETCDDRKEYMNQWSCGSARTMKRGYKNDNYELCKYNGVDVKPTTVRSNSSKLDRHRQPRGWEQLYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLEERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIRNGALLRVYVPRSSLPGFYRTSLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLETILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYASQ PGKPPRKDEL
>DBL1-ID2aFCR3-PE38(SEQ ID NO:67)
NHSDSGKYDPCEKKLPPYDDNDQWKCQQNSSDGSGKPENICVPPRRERLCTYNLENLKFDKIRDNNAFLADVLLTARNEGEKIVQNHPDTNSSNVCNALERSFADLADIIRGTDQWKGTNSNLEKNLKQMFAKIRENDKVLQDKYPKDQKYTKLREAWWNANRQKVWEVITCGARSNDLLIKRGWRTSGKSDRKKNFELCRKCGHYEKEVPTKLDYVPQFLRWLTEWIEDFYREKQNLIDDMERHREECTREDHKSKEGTSYCSTCKDKCKKYCECVKKWKTEWENQENKYKDLYEQNKNKTSQKNTSRYDDYVKDFFEKLEANYSSLENYIKGDPYFAEYATKLSFILNPSDANNPSGETANHNDEACNCNESGISSVGQAQTSGPSSNKTCITHSSIKTNKKKECKDVKLGVRENDKDLKICVIEDTSLSGVDNCCCQDLLGILQENCSDNKRGSSSNDSCDNKNQDECQKKLEKVFASLTNGYKCDKCKSGTSRSKKKWIWKKSSGNEEGLQEEYANTIGLPPRTQSLYLGNLPKLENVCEDVKDINFDTKEKFLAGCLIVSFHEGKNLKKRYPQNKNSGNKENLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQNNFGKLFGKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWTAMKHGAEMNITTCNADGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVENFCEQRQAKVKDVITNCKSCKESGNKCKTECKTKCKDECEKYKKFIEACGTAGGGIGTAGSPWSKRWDQIYKRYSKHIEDAKRNRKAGTKNCGTSSTTNAAASTDENKCVQSDIDSFFKHLIDIGLTTPSSYLSNVLDDNICGADKAPWTTYTTYTTTEKCNKERDKSKSQSSDTLVVVNVPSPLGNTPYRYKYACQCKIPTNEETCDDRKEYMNQWSCGSARTMKRGYKNDNYELCKYNGVDVKPTTVRSNSSKLDPEGGSLAALTAHQACHLPLETFTRHRQPRGWEQLEQCGYPVQRLVALYLAARLSWNQVDQVIRNALASPGSGGDLGEAIREQPEQARLALTLAAAESERFVRQGTGNDEAGAANGPADSGDALLERNYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLEERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIRNGALLRVYVPRSSLPGFYRTSLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLETILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYASQPGKPPRKDEL
>ID1-ID2a3D7-PE38(SEQ ID NO:68)
LSFILNSSDANNPSEKIQKNNDEVCNCNESGIASVEQEQISDPSSNKTCITHSSIKANKKKVCKHVKLGVRENDKDLRVCVIEHTSLSGVENCCCQDFLRILQENCSDNKSGSSSNGSCNNKNQEACEKNLEKVLASLTNCYKCDKCKSEQSKKNNKNWIWKKSSGKEGGLQKEYANTIGLPPRTQSLCLVVCLDEKGKKTQELKNIRTNSELLKEWIIAAFHEGKNLKPSHEKKNDDNGKKLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQKIFGKLFRKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWLAMKHGAGMNSTTCCGDGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVEHFCKQRQEKVKPVIENCKSCKESGGTCNGECKTECKNKCEVYKKFIEDCKGGDGTAGSSWVKRWDQIYKRYSKYIEDAKRNRKAGTKNCGPSSTTNAAENKCVQSDIDSFFKHLIDIGLTTPSSYLSIVLDDNICGADKAPWTTYTTYTTTEKCNKETDKSKLQQCNTAVVVNVPSPLGNTPHGYKYACQCKIPTNEETCDDRKEYMNQWSCGSARTMKRGYKNDNYELCKYNGVDVKPTTVRSNSSKLDPEGGSLAALTAHQACHLPLETFTRHRQPRGWEQLEQCGYPVQRLVALYLAARLSWNQVDQVIRNALASPGSGGDLGEAIREQPEQARLALTLAAAESERFVRQGTGNDEAGAANGPADSGDALLERNYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLEERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIRNGALLRVYVPRSSLPGFYRTSLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLETILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYASQPGKPPRKDEL
>ID1-ID2a3D7-PE38LR(SEQ ID NO:69)
LSFILNSSDANNPSEKIQKNNDEVCNCNESGIASVEQEQISDPSSNKTCITHSSIKANKKKVCKHVKLGVRENDKDLRVCVIEHTSLSGVENCCCQDFLRILQENCSDNKSGSSSNGSCNNKNQEACEKNLEKVLASLTNCYKCDKCKSEQSKKNNKNWIWKKSSGKEGGLQKEYANTIGLPPRTQSLCLVVCLDEKGKKTQELKNIRTNSELLKEWIIAAFHEGKNLKPSHEKKNDDNGKKLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQKIFGKLFRKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWLAMKHGAGMNSTTCCGDGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVEHFCKQRQEKVKPVIENCKSCKESGGTCNGECKTECKNKCEVYKKFIEDCKGGDGTAGSSWVKRWDQIYKRYSKYIEDAKRNRKAGTKNCGPSSTTNAAENKCVQSDIDSFFKHLIDIGLTTPSSYLSIVLDDNICGADKAPWTTYTTYTTTEKCNKETDKSKLQQCNTAVVVNVPSPLGNTPHGYKYACQCKIPTNEETCDDRKEYMNQWSCGSARTMKRGYKNDNYELCKYNGVDVKPTTVRSNSSKLDRHRQPRGWEQLYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLEERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIRNGALLRVYVPRSSLPGFYRTSLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLETILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYASQPGKPPRKDEL
>ID1-DBL2bFCR3-PE38LR(SEQ ID NO:70)
NYIKGDPYFAEYATKLSFILNPSDANNPSGETANHNDEACNCNESGISSVGQAQTSGPSSNKTCITHSSIKTNKKKECKDVKLGVRENDKDLKICVIEDTSLSGVDNCCCQDLLGILQENCSDNKRGSSSNDSCDNKNQDECQKKLEKVFASLTNGYKCDKCKSGTSRSKKKWIWKKSSGNEEGLQEEYANTIGLPPRTQSLYLGNLPKLENVCEDVKDINFDTKEKFLAGCLIVSFHEGKNLKKRYPQNKNSGNKENLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQNNFGKLFGKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWTAMKHGAEMNITTCNADGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVENFCEQRQAKVKDVITNCKSCKESGNKCKTECKTKCKDECEKYKKFIEACGTAGGGIGTAGSPWSKRWDQIYKRYSKHIEDAKRNRKAGTKNCGTSSTTNAAASTDENKCVQSDIDSFFKHLIDIGLTTPSSYLSNVLDDNICGADKAPWTTYTTYTTTEKCNKERDKSKSQSSDTLVVVNVPSPLGNTPYRYKYRHRQPRGWEQLYPTGAEFLGDGGDISFSTRGTQNWTVERLLQAHRQLEERGYVFVGYHGTFLEAAQSIVFGGVRARSQDLDAIWRGFYIAGDPALAYGYAQDQEPDARGRIRNGALLRVYVPRSSLPGFYRTSLTLAAPEAAGEVERLIGHPLPLRLDAITGPEEEGGRLETILGWPLAERTVVIPSAIPTDPRNVGGDLDPSSIPDKEQAISALPDYASQPGKPPRKDEL
>DT388,白喉毒素序列(SEQ ID NO:71)
MGADDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYDDDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKELGLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEINFETRGKRGQDAMYEYMAQACAGNRVRRSVGSSLSCINLDWDVIRDKTKTKIESLKEHGPIKNKMSESPNKTVSEEKAKQYLEEFHQTALEHPELSELKTVTGTNPVFAGANYAAWAVNVAQVIDSETADNLEKTTAALSILPGIGSVMGIADGAVHHNTEEIVAQSIALSSLMVAQAIPLVGELVDIGFAAYNFVESIINLFQVVHNSYNRPAY SPGHKTQPMHEF
与白喉毒素的截短片段融合的VAR2CSA多肽的序列
>DT388-DBL1-ID2a 3D7(SEQ ID NO:72)
MGADDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYDDDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKELGLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEINFETRGKRGQDAMYEYMAQACAGNRVRRSVGSSLSCINLDWDVIRDKTKTKIESLKEHGPIKNKMSESPNKTVSEEKAKQYLEEFHQTALEHPELSELKTVTGTNPVFAGANYAAWAVNVAQVIDSETADNLEKTTAALSILPGIGSVMGIADGAVHHNTEEIVAQSIALSSLMVAQAIPLVGELVDIGFAAYNFVESIINLFQVVHNSYNRPAYSPGHKTQPMHEFHSDSGTNDPCDRIPPPYGDNDQWKCAIILSKVSEKPENVFVPPRRQRMCINNLEKLNVDKIRDKHAFLADVLLTARNEGERIVQNHPDTNSSNVCNALERSFADIADIIRGTDLWKGTNSNLEQNLKQMFAKIRENDKVLQDKYPKDQNYRKLREDWWNANRQKVWEVITCGARSNDLLIKRGWRTSGKSNGDNKLELCRKCGHYEEKVPTKLDYVPQFLRWLTEWIEDFYREKQNLIDDMERHREECTSEDHKSKEGTSYCSTCKDKCKKYCECVKKWKSEWENQKNKYTELYQQNKNETSQKNTSRYDDYVKDFFKKLEANYSSLENYIKGDPYFAEYATKLSFILNSSDANNPSEKIQKNNDEVCNCNESGIASVEQEQISDPSSNKTCITHSSIKANKKKVCKHVKLGVRENDKDLRVCVIEHTSLSGVENCCCQDFLRILQENCSDNKSGSSSNGSCNNKNQEACEKNLEKVLASLTNCYKCDKCKSEQSKKNNKNWIWKKSSGKEGGLQKEYANTIGLPPRTQSLCLVVCLDEKGKKTQELKNIRTNSELLKEWIIAAFHEGKNLKPSHEKKNDDNGKKLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQKIFGKLFRKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWLAMKHGAGMNSTTCCGDGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVEHFCKQRQEKVKPVIENCKSCKESGGTCNGECKTECKNKCEVYKKFIEDCKGGDGTAGSSWVKRWDQIYKRYSKYIEDAKRNRKAGTKNCGPSSTTNAAENKCVQSDIDSFFKHLIDIGLTTPSSYLSIVLDDNICGADKAPWTTYTTYTTTEKCNKETDKSKLQQCNTAVVVNVPSPLGNTPHGYKYACQCKIPTNEETCDDRKEYMNQWSCGSARTMKRGYKNDNYELCKYNGVDVKPTTVRSNS SKLDSGR
>DT388-DBL1-ID2a FCR3(SEQ ID NO:73)
MGADDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYDDDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKELGLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEINFETRGKRGQDAMYEYMAQACAGNRVRRSVGSSLSCINLDWDVIRDKTKTKIESLKEHGPIKNKMSESPNKTVSEEKAKQYLEEFHQTALEHPELSELKTVTGTNPVFAGANYAAWAVNVAQVIDSETADNLEKTTAALSILPGIGSVMGIADGAVHHNTEEIVAQSIALSSLMVAQAIPLVGELVDIGFAAYNFVESIINLFQVVHNSYNRPAYSPGHKTQPMHEFHSDSGKYDPCEKKLPPYDDNDQWKCQQNSSDGSGKPENICVPPRRERLCTYNLENLKFDKIRDNNAFLADVLLTARNEGEKIVQNHPDTNSSNVCNALERSFADLADIIRGTDQWKGTNSNLEKNLKQMFAKIRENDKVLQDKYPKDQKYTKLREAWWNANRQKVWEVITCGARSNDLLIKRGWRTSGKSDRKKNFELCRKCGHYEKEVPTKLDYVPQFLRWLTEWIEDFYREKQNLIDDMERHREECTREDHKSKEGTSYCSTCKDKCKKYCECVKKWKTEWENQENKYKDLYEQNKNKTSQKNTSRYDDYVKDFFEKLEANYSSLENYIKGDPYFAEYATKLSFILNPSDANNPSGETANHNDEACNCNESGISSVGQAQTSGPSSNKTCITHSSIKTNKKKECKDVKLGVRENDKDLKICVIEDTSLSGVDNCCCQDLLGILQENCSDNKRGSSSNDSCDNKNQDECQKKLEKVFASLTNGYKCDKCKSGTSRSKKKWIWKKSSGNEEGLQEEYANTIGLPPRTQSLYLGNLPKLENVCEDVKDINFDTKEKFLAGCLIVSFHEGKNLKKRYPQNKNSGNKENLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQNNFGKLFGKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWTAMKHGAEMNITTCNADGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVENFCEQRQAKVKDVITNCKSCKESGNKCKTECKTKCKDECEKYKKFIEACGTAGGGIGTAGSPWSKRWDQIYKRYSKHIEDAKRNRKAGTKNCGTSSTTNAAASTDENKCVQSDIDSFFKHLIDIGLTTPSSYLSNVLDDNICGADKAPWTTYTTYTTTEKCNKERDKSKSQSSDTLVVVNVPSPLGNTPYRYKYACQCKIPTNEETCDDRKEYMNQWSCGSARTMKRGYKNDNYELCKYNGVDVKP TTVRSNSSKLDSGR
>DT388-ID1-ID2a 3D7(SEQ ID NO:74)
MGADDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYDDDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKELGLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEINFETRGKRGQDAMYEYMAQACAGNRVRRSVGSSLSCINLDWDVIRDKTKTKIESLKEHGPIKNKMSESPNKTVSEEKAKQYLEEFHQTALEHPELSELKTVTGTNPVFAGANYAAWAVNVAQVIDSETADNLEKTTAALSILPGIGSVMGIADGAVHHNTEEIVAQSIALSSLMVAQAIPLVGELVDIGFAAYNFVESIINLFQVVHNSYNRPAYSPGHKTQPMHEFLSFILNSSDANNPSEKIQKNNDEVCNCNESGIASVEQEQISDPSSNKTCITHSSIKANKKKVCKHVKLGVRENDKDLRVCVIEHTSLSGVENCCCQDFLRILQENCSDNKSGSSSNGSCNNKNQEACEKNLEKVLASLTNCYKCDKCKSEQSKKNNKNWIWKKSSGKEGGLQKEYANTIGLPPRTQSLCLVVCLDEKGKKTQELKNIRTNSELLKEWIIAAFHEGKNLKPSHEKKNDDNGKKLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQKIFGKLFRKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWLAMKHGAGMNSTTCCGDGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVEHFCKQRQEKVKPVIENCKSCKESGGTCNGECKTECKNKCEVYKKFIEDCKGGDGTAGSSWVKRWDQIYKRYSKYIEDAKRNRKAGTKNCGPSSTTNAAENKCVQSDIDSFFKHLIDIGLTTPSSYLSIVLDDNICGADKAPWTTYTTYTTTEKCNKETDKSKLQQCNTAVVVNVPSPLGNTPHGYKYACQCKIPTNEETCDDRKEYMNQWSCGSARTMKRGYKNDNYELCKYNGVDVKPTTVRSNSSKLDSGR
>DT388-ID1-ID2a FCR3(SEQ ID NO:75)
MGADDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYDDDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKELGLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEINFETRGKRGQDAMYEYMAQACAGNRVRRSVGSSLSCINLDWDVIRDKTKTKIESLKEHGPIKNKMSESPNKTVSEEKAKQYLEEFHQTALEHPELSELKTVTGTNPVFAGANYAAWAVNVAQVIDSETADNLEKTTAALSILPGIGSVMGIADGAVHHNTEEIVAQSIALSSLMVAQAIPLVGELVDIGFAAYNFVESIINLFQVVHNSYNRPAYSPGHKTQPMHEFNYIKGDPYFAEYATKLSFILNPSDANNPSGETANHNDEACNCNESGISSVGQAQTSGPSSNKTCITHSSIKTNKKKECKDVKLGVRENDKDLKICVIEDTSLSGVDNCCCQDLLGILQENCSDNKRGSSSNDSCDNKNQDECQKKLEKVFASLTNGYKCDKCKSGTSRSKKKWIWKKSSGNEEGLQEEYANTIGLPPRTQSLYLGNLPKLENVCEDVKDINFDTKEKFLAGCLIVSFHEGKNLKKRYPQNKNSGNKENLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQNNFGKLFGKYIKKNNTAEQDTSYSSLDELRESWWNTNKKYIWTAMKHGAEMNITTCNADGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVENFCEQRQAKVKDVITNCKSCKESGNKCKTECKTKCKDECEKYKKFIEACGTAGGGIGTAGSPWSKRWDQIYKRYSKHIEDAKRNRKAGTKNCGTSSTTNAAASTDENKCVQSDIDSFFKHLIDIGLTTPSSYLSNVLDDNICGADKAPWTTYTTYTTTEKCNKERDKSKSQSSDTLVVVNVPSPLGNTPYRYKYACQCKIPTNEETCDDRKEYMNQWSCGSARTMKRGYKNDNYELCKYNGVDVKPTTVRSNSS KLDSGR
>id1id2a FCR3 3x Nglyc突变体(asn/N to Gln/Q)(SEQ ID NO:128)
NYIKGDPYFAEYATKLSFILNPSDANNPSGETANHNDEACNCNESGISSVGQAQTSGPSSqKTCITHSSIKTNKKKECKDVKLGVRENDKDLKICVIEDTSLSGVDNCCCQDLLGILQENCSDNKRGSSSNDSCDNKNQDECQKKLEKVFASLTNGYKCDKCKSGTSRSKKKWIWKKSSGNEEGLQEEYANTIGLPPRTQSLYLGNLPKLENVCEDVKDINFDTKEKFLAGCLIVSFHEGKNLKKRYPQNKNSGNKENLCKALEYSFADYGDLIKGTSIWDNEYTKDLELNLQNNFGKLFGKYIKKqNTAEQDTSYSSLDELRESWWNTNKKYIWTAMKHGAEMqITTCNADGSVTGSGSSCDDIPTIDLIPQYLRFLQEWVENFCEQRQAKVKDVITNCKSCKESGNKCKTECKTKCKDECEKYKKFIEACGTAGGGIGTAGSPWSKRWDQIYKRYSKHIEDAKRNRKAGTKNCGTSSTTNAAASTDENKCVQSDIDSFFKHLIDIGLTTPSSYLSNVLDDNICGADKAPWTTYTTYTTTEKCNKERDKSKSQSSDTLVVVNVPSPLGNTPYRYKYACQCKIPTNEETCDDRKEYMNQWSCGSARTMKRGYKNDNYELCKYNGVDVKPTTVRSNSSKLD
>SpyCatcher(SEQ ID NO:129)
GAMVDTLSGLSSEQGQSGDMTIEEDSATHIKFSKRDEDGKELAGATMELRDSSGKTISTWISDGQVKDFYLYPGKYTFVETAAPDGYEVATAITFTVNEQGQVTVNGK ATKGDAHI
>Spytag(SEQ ID NO:130)
AHIVMVDAYKPTK
>最小Spytag序列(SEQ ID NO:131)
AHIVMVDA
>CnaB2的β-链(K-Tag)(SEQ ID NO:132)
ATHIKFSKRD
SpyLigase(SEQ ID NO:133)
HHHHHHDYDGQSGDGKELAGATMELRDSSGKTISTWISDGQVKDFYLYPGKYTFVETAAPDGYEVATAITFTVNEQGQVTVNGKATKGGSGGSGGSGEDSATHI
>异肽Spy0128(SEQ ID NO:134)
TDKDMTITFTNKKDAE
>split-Spy0128(SEQ ID NO:135)
ATTVHGETVVNGAKLTVTKNLDLVNSNALIPNTDFTFKIEPDTTVNEDGNKFKGVALNTPMTKVTYTNSDKGGSNTKTAEFDFSEVTFEKPGVYYYKVTEEKIDKVPGVSYDTTSYTVQVHVLWNEEQQKPVATYIVGYKEGSKVPIQFKNSLDSTTLTVKKKVSGTGGDRSKDFNFGLTLKANQYYKASEKVMIEKTTKGGQAPVQTEASIDQLYHFTLKDGESIKVTNLPVGVDYVVTEDDYKSEKYTTNVEVSPQDGAVKNIAGNS TEQETSTDKDMTI
>SpyCatcher-ΔN(SEQ ID NO:136)
EDSATHIKFSKRDEDGKELAGATMELRDSSGKTISTWISDGQVKDFYLYPGKYTFVETAAPDGYEVATAITFTVNEQGQVTVNGKATKGDAHI
>反向的Spytag序列(SEQ ID NO:137)
KTPKYADVMVIHA
实施例
方法
方法1-克隆和蛋白质表达
使用特异性引物(表2)，从密码子优化的FCR3(GenBank登录号GU249598)或3D7(GenBank登录号JQ247428)VAR2CSA基因扩增VAR2CSA序列片段。在一步PCR中扩增简单片段。在两步 PCR中制备氨基酸取代构建体。第一个PCR扩增了来自密码子优化的FCR3模板的两个片段，其含有重叠的互补末端。第二个PCR扩增了总构建体，使用两个重叠片段作为模板，使用特异于外部边界的引物。对所有片段进行测序以进行验证。将片段克隆到杆状病毒载体 pAcGP67-A(BD Biosciences)(被修饰成在C-末端含有V5和His 标签)或pEt大肠杆菌表达载体中。蛋白质在杆状病毒感染的昆虫细胞中表达作为分泌到细胞培养上清液的可溶性蛋白质或在大肠杆菌 Shuffle细胞中表达。使用用于基因设计的标准算法设计合成基因，考虑了二级DNA和RNA结构、重复区域和给定表达细胞系统的密码子选择。合成基因被制备用于Spy-tag和Spy-catcher融合以及CAR设计。
方法2-蛋白纯化和SDS-PAGE
使用交叉流(GE Healthcare)透析含有分泌的重组蛋白的经过滤的上清液。透析在10mM NaH2PO4(pH 7.4，Sigma-Aldrich) 和500mM NaCl中进行。将所得溶液过滤(0.2μm)，加入咪唑至终浓度为15mM。然后将蛋白质在1ml HisSelect柱(H8286， Sigma-Aldrich)上纯化。结合的蛋白质用10mM NaH2PO4(pH 7.4)， 500mM NaCl和500mM咪唑洗脱。通过使用20mM Tris(pH8)和 200mM NaCl中的HiLoad 16/60 Superdex 200柱(GEHealthcare) 的尺寸排阻色谱法进一步纯化石英晶体微量天平测量和SAXS所需的蛋白质以获得单体。通过SDS-PAGE验证蛋白质的纯度和结构完整性。
方法3-ELISA
将Falcon微量滴定板(351172，BD Biosciences)以3μg/ml的 CSPG(牛)(D8428，Sigma)或HSPG(H4777，Sigma)和100μg/ml 的CSA(C9819，Sigma)，CSC(400675，Seikagaku)和CSB(C3788， Sigma)在4℃孵育过夜。然后用振荡器在37℃下用TSM结合缓冲液 (20mMTris,150mM NaCl,2mM CaCl2,0.05％Tween-20,1％BSA, PH7.4，25℃)封闭板2小时。在TSM结合缓冲液中制备蛋白质的2 倍稀释系列(1.56mM-100mM)，并加入至板中，将其在37℃下在振荡器上孵育1小时。所有测量一式三份进行。将板在TSM洗涤缓冲液(20mM Tris,150mMNaCl,2mM CaCl2,0.05％Tween-20,PH7.4， 25℃)中洗涤三次。然后将板与1:3000的抗V5-HRP抗体(R96125， Invitrogen)一起在TSM结合缓冲液中于37℃在振荡器上孵育1小时。将板在TSM洗涤缓冲液中洗涤三次。最后用邻苯二胺底物(DAKO) 显影板15分钟。将反应用2.5M H2SO4淬灭。在490nm处测量吸光度。
方法4-石英晶体微量天平(Attana A100)
在Attana A100(Attana AB)上进行实验，使用镀金10MHz、 AT-切割石英晶体，聚苯乙烯芯片(3611-3103 Attana AB)。将所有缓冲液和试剂过滤至0.2μm。配体是以100μg/ml的浓度包被的CSPG (牛)(D8428，Sigma)或HSPG(H4777，Sigma)。通过加入配体溶液并在室温下静置孵育30分钟进行包被。随后用含有0.1％无Ig 的BSA(BSA-50，Rockland)的PBS封闭平板，在室温下进行30分钟。在每次实验之前，使用制造商预先定义的日常洗涤程序，将Attana A100用1％SDS洗涤。洗涤后，将运行缓冲液以25微升/分钟的流速在25℃下交换为PBS，并使机器稳定在0.5Hz/min的最大频率变化。经稳定后，PBS被注射多次以显示注射过程对基线影响最小。在注射样品前，将PBS作为空白注射。从最低浓度开始，以1:3稀释系列 (0.25μg/ml-60μg/ml)注射分析物。缔合时间设置为84秒，解离为5 分钟。由于结合的高亲和力，注射后不可能再生结合表面。收集的数据在Attester评估软件(Attana AB)中进行处理。曲线在简单的1： 1模型中拟合。通过曲线拟合确定kon和koff，并且基于KD＝koff/kon计算KD。
方法5-癌细胞结合测定
流式细胞术(FCM)用于检测VAR2CSA最小结合多肽对各种细胞系表面表达的CSPG的反应性。细胞在补充有10％胎牛血清(CHO 细胞，C32)，Hams F12(BeWo)的RPMI中培养，在37℃下保持在5％二氧化碳中，或从CPD缓冲液中的人血液样品(红细胞)中纯化。将细胞的等分试样(1×105)在+4℃在黑暗中平稳搅拌的情况下依次暴露于在FACS2(PBS+2％FCS)中稀释的VAR2CSA最小结合多肽(150,75或37nM)和α-V5-FITC(1:800)(Invitrogen)持续30分钟。作为阴性对照，使用截短形式的最小结合多肽和FACS2 缓冲液。基于正向和侧向散射信号门控完整细胞。使用FC500流式细胞仪(Beckman Coulter)从至少5000个细胞获得数据。在单个测定中处理和分析与特定细胞系相关的所有样品。
方法6-癌细胞结合测定
作为上述流式细胞术测定的替代方案，将细胞与在HBSS中稀释的VAR2CSA最小结合多肽和α-V5-FITC(1：500)(Invitrogen) 一起孵育。以与上述相同的浓度使用VAR2CSA多肽。在α-V5-FITC 染色后，将细胞在HBSS中洗涤3次，收集在无酶细胞分离缓冲液(Invitrogen)中，并在FACS Calibur(BD Biosciences)上进行FL-1 信号强度的分析。
缩写：CIDR，半胱氨酸丰富的域间区域；CSA，硫酸软骨素A； CSPG，硫酸软骨素蛋白聚糖；DBL，Duffy结合样结构域；FCM，流式细胞术；FV2，无N末端片段的VAR2CSA蛋白的全长胞外结构域；HSPG，硫酸乙酰肝素蛋白聚糖；ID，域间；IE，恶性疟原虫感染的红细胞；NTS，N末端区段；PM，胎盘疟疾；PfEMP1，恶性疟原虫红细胞膜蛋白1；PM，胎盘疟疾。
方法7-石蜡包埋的人体组织样品的染色
使用免疫组织化学(IHC)研究重组VAR2CSA与从人类患者获得的原发性癌症组织的结合。将在未进行抗原恢复的载玻片上点样的石蜡包埋组织在室温下与0.1-500nM V5-VAR2CSA变体或V5-对照蛋白(DBL4)孵育1小时，洗涤8分钟，用1:700小鼠抗-V5抗体孵育 30分钟，洗涤8分钟。随后使用Ventana Discovery XT平台利用 UltraMap抗小鼠HRP检测结合的抗V5。
实施例1截短的重组VAR2CSA蛋白质的产生
根据先前定义的结构域边界产生所有蛋白质截短体( M, JorgensenLM,Nielsen MA,Clausen TM,Ditlev SB,等人J Biol Chem 286:15908-15917)。为了简化起见，我们将CIDRPAM结构域划分为两个结构域ID2a和ID2b，其中ID2a是不包含类似CIDR的序列的 CIDRPAM的N端部分，ID2b对应于类似CIDR的序列。我们还使用了一个新的DBL2X边界，其结合了ID2a的93个氨基酸。为了简化起见，我们将这个边界称为DBL2Xb，而旧的边界称为DBL2Xa。用于克隆的引物列于表2中。如方法1中所述，片段在杆状病毒感染的昆虫细胞中表达为可溶性蛋白质。大多数蛋白质是基于FCR3基因型产生的。一些FCR3片段没有表达，而是基于3D7基因型进行制备的。蛋白质在分析中可互换使用，因为我们表明来自两种基因型的重组 VAR2CSA均等地与CSA结合。当通过SDS-PAGE比较还原和非还原样品时，所有蛋白质都显示出凝胶迁移率的变化(方法2)。这与分子内二硫键的形成是一致的。一些蛋白质通过非还原型SDS-PAGE 检测到形成高分子量复合物。这可能是由于在不成对的半胱氨酸之间形成分子间二硫键。这通过使用DTT将复合物还原成单体蛋白来证实。
实施例2来自FCR3和3D7的VAR2CSA以类似的亲和性和特异性结合CSA
与3D7或NF54(IE)相比，FCR3感染的红细胞(IE)在体外更强地粘附至CSA。如果差异与表达的VAR2CSA的序列差异相关，则该信息可用于定义粘附过程中涉及的残基。为了测试这一点，我们制作了一系列重叠的3D7 VAR2CSA片段，与我们以前针对FCR3测试的相同。
首先在固相结合测定(ELISA)(方法3中描述)中针对特异性 CSPG结合筛选蛋白质。然后通过尺寸排阻层析进一步纯化与CSPG 特异性结合的蛋白质，以获得纯单体蛋白质，并在石英晶体微量天平 (Attana A100)上进行动力学分析(分别为方法2和4)。在固相结合测定中，3D7 VAR2CSA片段显示出与其FCR3对应物非常相似的结合特征。在动力学分析中也是如此(表3)。传感图显示了在不同蛋白质浓度下收集的缔合和解离数据。这允许确定缔合速率常数 (kon)，解离速率常数(koff)和平衡常数(KD)。与峰值响应水平一起，这些参数给出了CSPG结合亲和力的估计。3D7和FCR3片段之间没有明显差异。一些片段显示出较低的亲和力，但是这个特征在片段对应物中得到保持。这表明3D7和FCR3VAR2CSA蛋白以相同的方式折叠并起作用。
表3：产生的VAR2CSA蛋白的CSA结合亲和力。亲和性作为使用石英晶体微量天平生物传感器(Attana A100)的动力学实验中确定的KD(nM)值给出。N/A：由于缺乏对CSA的结合，不能确定KD值的蛋白质。
* et al,JBC,2011中公开的蛋白
实施例3核心-CSA结合位点位于DBL2X结构域内
已经表明，VAR2CSA中的最小CSA结合区位于DBL2X-ID2b 内，需要侧翼结构域用于完全亲和结合( M,Jorgensen LM, Nielsen MA,Clausen TM,Ditlev SB,等人J Biol Chem 286: 15908-15917)。在这里我们分析了VAR2CSA的较短片段，以进一步绘制CSA结合所需的区域。
首先针对在ELISA中与CSA蛋白聚糖(CSPG)的结合筛选截短的蛋白质，然后进一步纯化以获得用于在石英晶体微量天平上检测的单体(分别为方法3、2和4)。最小结合区域为ID1-DBL2Xb(表 3)。该区域显示21.8nM的结合亲和力，其与全长VAR2CSA的结合亲和力相当。
胎盘IE对低硫酸化胎盘CSPG具有高选择性。它们不粘附到任何其它糖胺聚糖(GAG)，例如硫酸乙酰肝素(HS)。对于全长重组VAR2CSA蛋白也是如此。固相结合测定显示含有最小CSA结合区的VAR2CSA片段特异性结合CSA。为了证实这一点，进一步测试了最小结合片段与石英晶体微量天平上的硫酸乙酰肝素蛋白聚糖 (HSPG)的结合(方法4)。没有片段结合HSPG。
实施例4
VAR2CSA最小CSA结合区域特异性结合到一大组癌细胞
许多不同的癌细胞与蛋白聚糖CSPG4的高表达有关。该分子最初被描述为黑色素瘤的标志物，但最近已经发现许多癌症形式，包括癌症干细胞。与CSPG4附接的CS链已知主要是CSA。通过流式细胞仪分析最小的VAR2CSA片段之一(ID1-ID2a)与一大组各种各样的癌细胞系的结合(方法5和6)。非CSA结合蛋白ID1-DBL2Xa用作阴性对照。VAR2CSA重组蛋白(ID1-ID2a)以75nM强烈结合转录CSPG4的所有癌细胞系(微阵列数据)，包括皮肤黑素瘤(C32， MeWo)，肺癌(A549)，乳腺癌(HCC1395)，骨肉瘤(U2OS， MNNG/HOS)，横纹肌肉瘤(RH30)(表4和表5)。该蛋白质也与不表达CSPG4的皮肤T细胞淋巴瘤强烈结合(表4)。阴性对照蛋白ID1-DBL2Xa不结合任何测试的细胞系(表4)。此外，ID1-ID2a 与作为对照细胞的人红细胞没有相互作用。还分析了野生型和GAG 缺陷型中国仓鼠卵巢(CHO)细胞的ID1-ID2a相互作用。当分析 CHO-745细胞系(其中GAG合成被破坏)时，ID1-ID2a与野生型 CHO细胞的强烈相互作用被完全消除。通过将VAR2CSA与CSA， CSC或HS预混合抑制VAR2CSA与细胞结合来证实相互作用的CSA 特异性。CSC和HS对结合没有任何影响，而CSA有效地消除了VAR2CSA与癌细胞的结合。
根据这些结果，使用VAR2CSA的DBL1-ID2a或ID1-ID2a片段，通过流式细胞术筛选出较大组的癌细胞(表6和7)。该筛选的主要目的是鉴定适合体内异种移植建模的细胞系。
表4.使用VAR2CSA的最小结合结构域(ID1-ID2a)染色癌细胞系和阴性对照细胞。
细胞与单独的培养基(空白)或75nM的重组蛋白(ID1-DBL2 或ID1-ID2a)一起孵育30分钟，然后与1:800的抗V5-FITC (Invitrogen)孵育，细胞在每次孵育之间洗涤三次。显示了使用FC500 流式细胞仪(Becton Dickinson)从最少5000个细胞记录的平均FITC 荧光值。
表5.使用重组VAR2CSA的癌细胞系染色
将细胞与单独的培养基(空白)或75nM的重组蛋白(DBL1-ID2a 或ID1-ID2a)一起孵育30分钟，然后与1:800的抗V5-FITC (Invitrogen)孵育，细胞在每次孵育之间洗涤三次。显示了使用 HAL100Zeiss显微镜从最少4个高功率场图像记录的FITC荧光强度的中值评分。NS：无染色；+：弱；++：中等；+++：强；++++：非常强。
表6筛选用于结合重组VAR2CSA的多种人类癌细胞系(使用 DBL1-ID2a或ID1-ID2a)。
如方法5和6所述，通过流式细胞术测量结合。
NS：无染色；+：弱；++：中等；+++：强；++++：非常强。
表7用于结合重组VAR2CSA的更多人细胞癌细胞系的筛选(使用DBL1-ID2a或ID1-ID2a)
如方法5和6所述，通过流式细胞术测量结合。
显示的值是使用200nM的蛋白质浓度的平均荧光强度。
实施例5
重组VAR2CSA以高亲和力结合癌细胞
使用石英晶体微量天平生物传感器(Attana Cell200)分析重组 VAR2CSA片段DBL1-ID2a对癌细胞系、C32黑素瘤和两个Cho细胞系(实施例8中所述)的结合亲和力。针对与细胞表面的结合分析蛋白质的2倍稀释系列(25-400nM)，其中在每个新蛋白质注射之间再生结合表面。估计结合亲和力在纳摩尔范围内(表8)，其与对纯受体的结合亲和力相似(表3)。
表8.重组DBL1-ID2a(大肠杆菌)对表达CSA(C32和Cho WT) 和缺乏与CSA阴性细胞系(Cho 745)的结合的癌细胞的估计结合亲和力(KD)
N/A：由于缺乏对细胞的结合，KD无法确定
实施例6
重组VAR2CSA蛋白以高特异性结合癌组织
使用免疫组织化学(IHC)研究重组VAR2CSA与从人类患者获得的原发性癌症组织的结合。该方法采用人胎盘组织为阳性对照，扁桃体和肝组织为阴性对照。在VentanaDiscovery XT平台上优化了染色方案，没有进行表位修复。将贴在玻片上的石蜡包埋组织在室温下与0.1-500nM V5-VAR2CSA(ID1-ID2a)或V5-对照蛋白(DBL4) 孵育1小时，洗涤8分钟，与1：700小鼠抗-V5抗体孵育30分钟，洗涤8分钟。随后使用UltraMap抗小鼠HRP检测结合的抗V5。 V5-VAR2CSA以0.5nM浓度染色人胎盘，在扁桃体或正常肝脏中无染色。通过向反应缓冲液中加入200μg/μl CSA可以完全阻断染色。 V5-对照蛋白质不会在任何测试浓度下染色人胎盘组织。当用1nM V5-VAR2CSA染色时，代表24个正常器官的多器官组织微阵列(TMA)显示低或无染色，而乳腺，结肠，直肠，前列腺，肾，肝，膀胱，胰腺，鳞状细胞，肺，胆囊，胃，睾丸，卵巢，子宫，肾上腺，甲状腺和胸腺，造血系统和结缔组织(肉瘤)的癌样品阳性染色，强度等于或高于人胎盘阳性对照组织(表9)。
表9.使用重组VAR2CSA检测原代人肿瘤样品上的CSA。表格显示了实施例10中所述的主要癌症组所染色的阳性数/病例总数。阳性染色定义为等于或高于胎盘组织中观察到的强度。
实施例7
鉴定由VAR2CSA靶向的潜在CSPG分子
筛选具有V5-标签的重组VAR2CSA蛋白(DBL1-ID2a)与一组转染的表达>3000个人膜受体的HEK293细胞的结合。已经鉴定了一组25个受体作为VAR2CSA的潜在靶标(表10)。VAR2CSA与这些受体之间的相互作用将通过在HEK293系统和ELISA中通过CSA 和HS的抑制进行结合特异性的分析来进一步验证。
表10.实验鉴定为VAR2CSA潜在靶标的受体。
实施例8
双特异性VAR2CSA-scFv-抗CD3蛋白
已经将DBL1-ID2a和ID1-ID2a VAR2CSA基因片段与编码针对 CD3的单链抗体的scFv序列融合。这些VAR2CSA-scFv蛋白在大肠杆菌或昆虫细胞中表达。这些VAR2CSA-scFv蛋白针对表达CSA的癌细胞和表达CD3的T细胞是双特异性的。因此，这些蛋白质能够通过VAR2CSA结构域结合肿瘤环境中的CSA，同时通过scFv-抗CD3 结构域募集CD3阳性T细胞。
实施例9
VAR2CSA-Spy融合蛋白
已经将DBL1-ID2a和ID1-ID2a VAR2CSA基因片段融合到 SpyTag、K-tag、SpyCatcher和SpyCather-ΔN(SEQ ID NO： 130,129,136)的各种变体。这些VAR2CSA-Spy蛋白在大肠杆菌或昆虫细胞中表达。重组VAR2CSA-Spy蛋白可以与相应的分割蛋白结合配偶体形成异肽键。例如，VAR2CSA-SpyTag可以结合任何具有 SpyCatcher或ΔN-SpyCather序列的分子；VAR2CSA-SpyCatcher或 VAR2CSA-ΔN-SpyCather可以与任何具有SpyTag序列的分子结合； VAR2CSA-SpyTag可以在SpyLigase存在下与任何具有K-Tag序列的分子结合；或VAR2CSA-K-Tag可以在SpyLigase存在下与任何具有K-Tag序列的分子结合。此外，VAR2CSA-K-Tag可以在SpyLigase 存在下通过K-Tag聚合(彼此连接)，并随后与任何蛋白质表达SpyCatcher(如Spy-CAR)组合。因此，这些蛋白质能够结合具有分割蛋白结合系统的相应序列的任何蛋白质序列。通过将Spy标记的 VAR2CSA蛋白与Spy-catcher或Spy-catcher标记的VAR2CSA与 Spy-tag简单混合，我们观察到蛋白质之间的快速和有效的偶联，如通过尺寸排阻色谱法或SDS page所测量的。将Spy标记的VAR2CSA 注射到具有建立的肿瘤的小鼠中，随后IV注射荧光标记的 Spy-catcher，通过IVIS体内成像和死后分析证明，Spy-tag和Spy-catcher的异肽键和配对确实在体内发生。
实施例10
双特异性VAR2CSA-Spy-scFv-抗CD3融合蛋白
已经将DBL1-ID2a和ID1-ID2a VAR2CSA基因片段与SpyTag、 K-tag、SpyCatcher和SpyCather-ΔN(SEQ ID NO：130,129,136)和编码针对CD3的单链抗体的scFv序列的各种变体融合。这些 VAR2CSA-Spy蛋白在大肠杆菌或昆虫细胞中表达。重组 VAR2CSA-Spy-scFv-抗-CD3融合蛋白可以与相应的分割蛋白结合配偶体形成异肽键。例如，VAR2CSA-Spy-scFv-抗-CD3可以结合任何具有SpyCatcher或ΔN-SpyCather序列的分子； VAR2CSA-SpyCatcher-scFv-抗-CD3或 VAR2CSA-ΔN-SpyCather-scFv-抗-CD3可以结合任何具有SpyTag序列的分子；VAR2CSA-Spy-scFv-抗-CD3可以在SpyLigase存在下与任何具有K-Tag序列的分子结合；或VAR2CSA-K-Tag-scFv抗CD3可以在SpyLigase存在下与任何具有K-标记序列的分子结合。此外， VAR2CSA-K-Tag-scFv-抗-CD3可以在SpyLigase存在下通过K-Tag聚合(彼此连接)，并随后与任何蛋白质表达SpyCatcher(例如 Spy-CAR)组合。如实施例9所述合成和验证VAR2CSA-Spy-scFv- 抗-CD3蛋白。
实施例11
表达VAR2CSA(VAR2-CAR)的嵌合抗原受体
使用密码子优化算法(Optimizer，PMID：17439967)合成CAR 编码序列，并亚克隆到编码第二代(CD28跨膜和细胞内T细胞信号传导结构域和CD3-ζ链胞内T细胞信号传导结构域)；或第三代(与 CD8，4-1BB和CD3-ζ细胞内T细胞信号传导结构域连接的CD28跨膜结构域)的“目的”载体中。通过用编码CAR逆转录病毒载体的质粒和RD114包膜糖蛋白转染293GP包装细胞(Clonetech)，48-72 小时后收集培养上清液(s/n)产生逆转录病毒载体上清液。将培养上清液冷冻或立即用于将OKT3和IL-2活化的人PBMC转导连续2天 (在用预先暴露于含有s/n的载体的稀释液的RECTRONECTIN (Clonetech)包被的平板上培养淋巴细胞)。通过流式细胞术测定转导的T细胞上的CAR表达。(表11)。
表11.通过流式细胞术检测转导的T细胞的质膜上的VAR2-CAR 的表达。
阳性细胞百分比 阴性细胞百分比
VAR2-CAR 61.40％ 38.60％
实施例12
表达Spy-CAR的嵌合抗原受体
使用密码子优化算法(Optimizer，PMID：17439967)合成CAR 编码序列SpyTag，SpyCatcher，ΔN-SpyCatcher或K-Tag(SEQ ID NO：130，129，136或132)，并亚克隆到编码第二代(CD28跨膜和细胞内T细胞信号传导结构域和CD3-ζ链胞内T细胞信号转导结构域)；或第三代(与CD28，4-1BB和CD3-ζ细胞内T细胞信号传导结构域连接的CD8跨膜结构域)的“目的”载体。通过用编码CAR逆转录病毒载体和RD114包膜糖蛋白的质粒转染293GP包装细胞(Clonetech)，48-72小时后收集培养上清液(s/n)产生逆转录病毒载体上清液。将培养上清液冷冻或立即用于将OKT3和IL-2活化的人PBMC转导连续2天(在用预先暴露于含有s/n的载体的稀释液的 RECTRONECTIN(Clonetech)包被的平板上培养淋巴细胞)。通过流式细胞术测定转导的T细胞上的CAR表达(表12)。
表12.通过流式细胞术检测的转导的T细胞的质膜上的Spy-CAR 的表达。
阳性细胞百分比 阴性细胞百分比
Spy-CAR 64.82％ 35.18％
实施例13
表达Spy-CAR并随后与VAR2-Spy融合蛋白连接的嵌合抗原受体：VAR2-Spy-Spy-CAR
使用密码子优化算法(Optimizer，PMID：17439967)合成CAR 编码序列SpyTag，SpyCatcher，ΔN-SpyCatcher或K-标签(SEQ ID NO：130，129，136或132)，并亚克隆到编码第二代(CD28跨膜和细胞内T细胞信号传导结构域和CD3-ζ链细胞内T细胞信号转导结构域)；或第三代(与CD28，4-1BB和CD3-ζ细胞内T细胞信号结构域连接的CD8跨膜结构域)的“目的”载体。通过用编码CAR逆转录病毒载体和RD114包膜糖蛋白的质粒转染293GP包装细胞 (Clonetech)，48-72小时后收集培养上清液(s/n)产生逆转录病毒载体上清液。将培养物上清液冷冻或立即用于转导OKT3和IL-2活化的人PBMC连续2天(在用预先暴露于含有s/n的载体的稀释液的 RECTRONECTIN(Clonetech)包被的平板上培养淋巴细胞)。 Spy-CAR转基因T细胞随后与相应的VAR2CSA-Spy融合蛋白(对应于Spy-CAR的分割蛋白结合系统的另一半)孵育以产生完全功能的 VAR2CSA-Spy-Spy-CAR受体。通过流式细胞术和/或共同免疫荧光测定转导的T细胞上的CAR表达(表13)。
表13.随后将Spy-CAR转基因T细胞与相应的VAR2CSA-Spy融合蛋白(对应于Spy-CAR的分割蛋白结合系统的另一半)孵育以产生武装的VAR2CSA-Spy-Spy-CAR受体，如通过流式细胞术和免疫荧光(IF)共定位测量的。
实施例14
比较和选择VAR2-CAR
为了针对VAR2-CAR和VAR2-Spy-Spy-CAR的转导效率进行归一化，分析效应T细胞的百分比CAR表达(转导百分比)。基于此，计算效应细胞与靶细胞的比率(E:T比率)。这与％裂解进行比较。一个裂解单位被定义为以10:1的E:T比率的靶细胞的30％裂解(如专利申请WO 2013059593 A1中所定义)。这个功能“单位”定义量化了所比较的每个转导的效应细胞群体中裂解活性的量。裂解单位表示构建体之间转导效率差异的归一化E：T比值。
实施例15
不同VAR2-CAR的裂解活性
为了比较不同VAR2-CAR构建体的裂解潜力，使用五种不同的表达CSA的肿瘤细胞系(例如Colo205结肠直肠癌，REH白血病， UM-UC13膀胱癌，PC-3前列腺癌和RH30儿科横纹肌肉瘤)在5lCr 释放CTL测定或比色细胞毒性测定中比较编码正向和反向翻译的 VAR2CSA的CAR。作为对照，还在非表达CSA的非致瘤性HEK293 细胞上测试VAR2-CAR。根据实施例14所述的各个CAR构建体的转导百分比，将E：T比率归一化，因此裂解值是直接可比的。VAR2-CAR 的裂解活性的一个实例示于表14中。
实施例16
不同VAR2-Spy-Spy-CAR的裂解活性
为了比较不同VAR2-Spy-Spy-CAR构建体的裂解潜力，使用五种不同的表达CSA的肿瘤细胞系(例如Colo205结肠直肠癌，REH白血病，UM-UC13膀胱癌，PC-3前列腺癌和RH30儿科横纹肌肉瘤) 在5lCr释放CTL测定或比色细胞毒性测定中比较编码正向和反向翻译的VAR2-Spy-Spy-CAR构建体的CAR。作为对照，还在非表达CSA 的非致瘤性HEK293细胞上测试VAR2-Spy-Spy-CAR。根据实施例 14所述的各个CAR构建体的转导百分比，将E：T比率归一化，因此裂解值是直接可比的。VAR2-Spy武装的Spy-CAR的裂解活性的一个实例示于表14中。
表14.用VAR2-CAR或Spy-CAR纯化和转导随后用VAR2-Spy 武装T细胞，如实施例13所示。将CAR以指定的E：T比率加入到 UC13细胞中，T细胞介导的细胞毒性为如实施例15和16所述测量。
实施例17
不同VAR2-CAR和VAR2-Spy-Spy-CAR在体内的裂解活性
在第0天，用被改造以表达荧光素酶的CSA阳性的人癌细胞系(例如Colo205结肠直肠癌，REH白血病，UM-UC13膀胱癌，PC-3前列腺癌和RH30小儿横纹肌肉瘤)注射NSG(NODscidγ)免疫缺陷小鼠。在第3天，用1x 107个对照T细胞(“模拟组”，未转导)或用 VAR2-CAR和VAR2-Spy-Spy-CAR转导的1x 107个T细胞处理小鼠。使用IVIS的生物发光成像在30天的时间段内测量肿瘤负荷。用3mg D-荧光素(Caliper Life Sciences，Inc。)腹膜内(i.p.)注射小鼠，注射后4分钟，麻醉的小鼠以10-60秒的暴露时间成像。LIVING IMAGE软件用于将每只小鼠的生物发光信号分析为光子/s/cm/sr。
附加说明
本文引用的所有参考文献，包括出版物，专利申请和专利通过引用并入本文，其程度如同每个参考文献被单独地和具体地指示通过引用并入和在本文以其整体示出。
在描述本发明的上下文中(特别是在所附权利要求的上下文中) 中使用术语“一个”和“一种”和“所述”以及类似的指示物应被解释为包括单数和复数，除非本文另有说明或明显与上下文相矛盾。术语“包含”，“具有”，“含有”和“包括”将被解释为开放式术语(即意义“包括但不限于”)，除非另有说明。除非本文另有说明，本文中值的范围的描述仅旨在用作单独提及落在该范围内的每个单独值的速记方法，并且将每个单独的值并入本说明书中，如同在本文中单独列举一样。本文所述的所有方法可以以任何合适的顺序进行，除非本文另有说明或以其它方式明确地与上下文相矛盾。本文提供的任何和所有示例或示例性语言(例如“例如”)的使用仅旨在更好地举例说明本发明，并且不对本发明的范围构成限制，除非另有说明。说明书中的任何语言不应被解释为指示任何未被要求保护的元素对于本发明的实践是必需的。
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Val Thr Thr Thr Glu Asn Lys Cys Val Gln Ser Asp Ile Asp Ser Phe
500 505 510
Phe Lys His Leu Ile Asp Ile Gly Leu Thr Thr Pro Ser Ser Tyr Leu
515 520 525
Ser Ile Val Leu Asp Asp Asn Ile Cys Gly Asp Asp Lys Ala Pro Trp
530 535 540
Thr Thr Tyr Thr Thr Tyr Thr Thr Tyr Thr Thr Thr Glu Lys Cys Asn
545 550 555 560
Lys Glu Arg Asp Lys Ser Lys Ser Gln Gln Ser Asn Thr Ser Val Val
565 570 575
Val Asn Val Pro Ser Pro Leu Gly Asn Thr Pro His Gly Tyr Lys Tyr
580 585 590
Ala Cys Gln Cys Lys Ile Pro Thr Asn Glu Glu Thr Cys Asp Asp Arg
595 600 605
Lys Glu Tyr Met Asn Gln Trp Ile Ser Asp Thr Ser Lys Asn Pro Lys
610 615 620
Gly Ser Gly Ser Thr Asn Asn Asp Tyr Glu Leu Tyr Thr Tyr Asn Gly
625 630 635 640
Val Lys Glu Thr Lys Leu Pro Lys Lys Leu Asn Ser Pro Lys Leu Asp
645 650 655
<210> 4
<211> 643
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 4
Asp Tyr Ile Lys Asp Asp Pro Tyr Ser Lys Glu Tyr Thr Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Ser Ser Asp Ala Asn Thr Ser Ser Gly Glu Thr
20 25 30
Ala Asn His Asn Asp Glu Ala Cys Asn Cys Asn Glu Ser Glu Ile Ser
35 40 45
Ser Val Gly Gln Ala Gln Thr Ser Gly Pro Ser Ser Asn Lys Thr Cys
50 55 60
Ile Thr His Ser Phe Ile Lys Ala Asn Lys Lys Lys Val Cys Lys Asp
65 70 75 80
Val Lys Leu Gly Val Arg Glu Asn Asp Lys Val Leu Arg Val Cys Val
85 90 95
Ile Glu Asp Thr Ser Leu Ser Gly Val Asp Asn Cys Cys Cys Gln Asp
100 105 110
Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Arg Gly Ser
115 120 125
Ser Ser Asn Gly Ser Cys Asn Asn Lys Asn Gln Asp Glu Cys Gln Lys
130 135 140
Lys Leu Glu Lys Val Phe Val Ser Leu Thr Asn Gly Tyr Lys Cys Asp
145 150 155 160
Lys Cys Lys Ser Gly Thr Ser Thr Val Asn Lys Lys Trp Ile Trp Lys
165 170 175
Lys Ser Ser Gly Asn Glu Lys Gly Leu Gln Lys Glu Tyr Ala Asn Thr
180 185 190
Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu Gly Asn Leu Pro
195 200 205
Lys Leu Gly Asn Val Cys Glu Asp Val Thr Asp Ile Asn Phe Asp Thr
210 215 220
Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Ala Ala Phe His Glu Gly
225 230 235 240
Lys Asn Leu Lys Ile Ser His Glu Lys Lys Lys Gly Asp Asn Gly Lys
245 250 255
Lys Leu Cys Lys Ala Leu Glu Tyr Ser Phe Ala Asp Tyr Gly Asp Leu
260 265 270
Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp Leu Glu
275 280 285
Leu Asn Leu Gln Lys Ala Phe Gly Lys Leu Phe Gly Lys Tyr Ile Lys
290 295 300
Lys Asn Ile Ala Ser Asp Glu Asn Thr Ser Tyr Ser Ser Leu Asp Glu
305 310 315 320
Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Thr Ala
325 330 335
Met Lys His Gly Ala Glu Met Asn Ser Thr Met Cys Asn Ala Asp Gly
340 345 350
Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp Ile Pro Thr Thr Asp
355 360 365
Phe Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His Phe
370 375 380
Cys Lys Gln Arg Gln Glu Lys Val Asn Ala Val Ile Glu Asn Cys Asn
385 390 395 400
Ser Cys Lys Asn Thr Ser Gly Glu Arg Lys Ile Gly Gly Thr Cys Asn
405 410 415
Gly Asp Cys Lys Thr Glu Cys Lys Asn Lys Cys Glu Ala Tyr Lys Asn
420 425 430
Phe Ile Glu Asp Cys Lys Gly Gly Asp Gly Thr Ala Gly Ser Ser Trp
435 440 445
Val Lys Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys His Ile Glu
450 455 460
Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys Ser Cys Gly Pro Ser
465 470 475 480
Ser Ile Thr Asn Ala Ser Val Ser Thr Asp Glu Asn Lys Cys Val Gln
485 490 495
Ser Asp Ile Asp Ser Phe Phe Lys His Leu Ile Asp Ile Gly Leu Thr
500 505 510
Thr Pro Ser Ser Tyr Leu Ser Ile Val Leu Asp Glu Asn Asn Cys Gly
515 520 525
Glu Asp Asn Ala Pro Trp Thr Thr Tyr Thr Thr Tyr Thr Thr Thr Glu
530 535 540
Lys Cys Asn Lys Asp Lys Lys Lys Ser Lys Ser Gln Ser Cys Asn Thr
545 550 555 560
Ala Val Val Val Asn Val Pro Ser Pro Leu Gly Asn Thr Pro His Glu
565 570 575
Tyr Lys Tyr Ala Cys Gln Cys Lys Ile Pro Thr Thr Glu Glu Thr Cys
580 585 590
Asp Asp Arg Lys Glu Tyr Met Asn Gln Trp Ile Ser Asp Thr Ser Lys
595 600 605
Lys Gln Lys Gly Ser Gly Ser Thr Asn Asn Asp Tyr Glu Leu Tyr Thr
610 615 620
Tyr Thr Gly Val Lys Glu Thr Lys Leu Pro Lys Lys Leu Asn Ser Pro
625 630 635 640
Lys Leu Asp
<210> 5
<211> 640
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 5
Ser Tyr Val Lys Asn Asp Pro Tyr Ser Lys Glu Tyr Val Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Pro Ser Asp Ala Asn Asn Pro Ser Gly Glu Thr
20 25 30
Ala Asn His Asn Asp Glu Ala Cys Asn Pro Asn Glu Ser Glu Ile Ala
35 40 45
Ser Val Gly Gln Ala Gln Thr Ser Asp Arg Leu Ser Gln Lys Ala Cys
50 55 60
Ile Thr His Ser Phe Ile Gly Ala Asn Lys Lys Ile Val Cys Lys Asp
65 70 75 80
Val Lys Leu Gly Val Arg Glu Lys Asp Lys Asp Leu Lys Ile Cys Val
85 90 95
Ile Glu Asp Asp Ser Leu Arg Gly Val Glu Asn Cys Cys Phe Lys Asp
100 105 110
Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Ser Gly Ser
115 120 125
Ser Ser Asn Gly Ser Cys Asn Asn Lys Asn Gln Asp Glu Cys Gln Lys
130 135 140
Lys Leu Asp Glu Ala Leu Ala Ser Leu His Asn Gly Tyr Lys Cys Asp
145 150 155 160
Lys Cys Lys Ser Gly Thr Ser Arg Ser Lys Lys Ile Trp Thr Trp Arg
165 170 175
Lys Phe Pro Gly Asn Gly Glu Gly Leu Gln Lys Glu Tyr Ala Asn Thr
180 185 190
Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu Gly Asn Leu Arg
195 200 205
Lys Leu Glu Asn Val Cys Lys Gly Val Thr Asp Ile Asn Phe Asp Thr
210 215 220
Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Ala Ala Phe His Glu Gly
225 230 235 240
Lys Asn Leu Lys Ile Ser Asn Lys Lys Lys Asn Asp Asp Asn Gly Lys
245 250 255
Lys Leu Cys Lys Asp Leu Lys Tyr Ser Phe Ala Asp Tyr Gly Asp Leu
260 265 270
Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp Leu Glu
275 280 285
Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe Arg Lys Tyr Ile Lys
290 295 300
Lys Asn Ile Ala Ser Asp Glu Asn Thr Leu Tyr Ser Ser Leu Asp Glu
305 310 315 320
Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Leu Ala
325 330 335
Met Lys His Gly Thr Thr Cys Ser Ser Gly Ser Gly Asp Asn Gly Asp
340 345 350
Gly Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp Met Ser Thr Ile
355 360 365
Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His
370 375 380
Phe Cys Lys Gln Arg Gln Glu Lys Val Lys Asp Val Ile Glu Asn Cys
385 390 395 400
Lys Ser Cys Lys Asn Thr Ser Gly Glu Arg Ile Ile Gly Gly Thr Cys
405 410 415
Gly Ser Asp Cys Lys Thr Lys Cys Lys Gly Glu Cys Asp Ala Tyr Lys
420 425 430
Asn Phe Ile Glu Glu Cys Lys Arg Gly Asp Gly Thr Ala Gly Ser Pro
435 440 445
Trp Ser Lys Arg Trp Asp Gln Ile Tyr Met Arg Tyr Ser Lys Tyr Ile
450 455 460
Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Thr
465 470 475 480
Ser Ser Thr Thr Asn Ala Ala Glu Asn Lys Cys Val Gln Ser Asp Ile
485 490 495
Asp Ser Phe Phe Lys His Leu Ile Asp Ile Gly Leu Thr Thr Pro Ser
500 505 510
Ser Tyr Leu Ser Ile Val Leu Asp Glu Asn Ile Cys Gly Asp Asp Lys
515 520 525
Ala Pro Trp Thr Thr Tyr Thr Thr Tyr Thr Thr Thr Glu Lys Cys Asn
530 535 540
Lys Glu Thr Asp Lys Ser Lys Ser Gln Ser Cys Asn Thr Ala Val Val
545 550 555 560
Val Asn Val Pro Ser Pro Leu Gly Asn Thr Pro His Gly Tyr Lys Tyr
565 570 575
Ala Cys Glu Cys Lys Ile Pro Thr Thr Glu Glu Thr Cys Asp Asp Arg
580 585 590
Lys Glu Tyr Met Asn Gln Trp Ile Ser Asp Thr Ser Lys Lys Pro Lys
595 600 605
Gly Gly Arg Ser Thr Asn Asn Asp Tyr Glu Leu Tyr Thr Tyr Asn Gly
610 615 620
Val Lys Glu Thr Lys Leu Pro Lys Lys Ser Ser Ser Ser Lys Leu Asp
625 630 635 640
<210> 6
<211> 358
<212> PRT
<213> 恶性疟原虫
<400> 6
Lys Cys Glu Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Asn Ile
1 5 10 15
Trp Ile Trp Arg Lys Phe Pro Gly Asn Gly Glu Gly Leu Gln Lys Glu
20 25 30
Tyr Ala Asn Thr Ile Gly Leu Pro Pro Arg Thr His Ser Leu Tyr Leu
35 40 45
Gly Asn Leu Pro Lys Leu Glu Asn Val Cys Lys Asp Val Lys Asp Ile
50 55 60
Asn Phe Asp Thr Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Ala Ala
65 70 75 80
Phe His Glu Gly Lys Asn Leu Lys Thr Thr Tyr Pro Gln Asn Lys Asn
85 90 95
Ala Asp Asn Asn Ser Lys Leu Cys Lys Asp Leu Lys Tyr Ser Phe Ala
100 105 110
Asp Tyr Gly Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Asp Phe
115 120 125
Thr Lys Asp Leu Glu Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe
130 135 140
Arg Lys Tyr Ile Lys Lys Asn Ile Ala Ser Asp Glu Asn Thr Leu Tyr
145 150 155 160
Ser Ser Leu Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys
165 170 175
Tyr Ile Trp Leu Ala Met Lys His Gly Ala Glu Met Asn Ser Thr Met
180 185 190
Cys Asn Gly Asp Gly Ser Val Thr Gly Ser Ser Asp Ser Gly Ser Thr
195 200 205
Thr Cys Ser Gly Asp Asn Gly Ser Ile Ser Cys Asp Asp Ile Pro Thr
210 215 220
Ile Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu
225 230 235 240
His Phe Cys Lys Gln Arg Gln Glu Lys Val Lys Pro Val Ile Glu Asn
245 250 255
Cys Lys Ser Cys Lys Asn Thr Ser Gly Glu Arg Ile Ile Gly Gly Thr
260 265 270
Cys Gly Ser Asp Cys Glu Lys Lys Cys Lys Gly Glu Cys Asp Ala Tyr
275 280 285
Lys Lys Phe Ile Glu Glu Cys Lys Gly Gly Gly Gly Gly Thr Gly Thr
290 295 300
Ala Gly Ser Pro Trp Ser Lys Arg Trp Asp Gln Ile Tyr Lys Arg Tyr
305 310 315 320
Ser Lys Tyr Ile Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys
325 330 335
Ser Cys Gly Pro Ser Ser Thr Thr Asn Ala Ala Ala Ser Thr Thr Glu
340 345 350
Ser Lys Cys Val Gln Ser
355
<210> 7
<211> 333
<212> PRT
<213> 恶性疟原虫
<400> 7
Lys Cys Asp Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Lys Asn
1 5 10 15
Trp Ile Trp Lys Gln Phe Pro Gly Asn Gly Glu Gly Leu Gln Lys Glu
20 25 30
Tyr Ala Asn Thr Ile Gly Leu Pro Pro Arg Thr His Ser Leu Tyr Leu
35 40 45
Gly Asn Leu Pro Lys Leu Glu Asn Val Cys Lys Gly Val Thr Asp Ile
50 55 60
Asn Phe Asp Thr Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Ala Ala
65 70 75 80
Phe His Glu Gly Lys Asn Leu Lys Thr Ser His Glu Lys Lys Lys Gly
85 90 95
Asp Asn Gly Lys Lys Leu Cys Lys Asp Leu Lys Tyr Ser Phe Ala Asp
100 105 110
Tyr Gly Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Asp Phe Thr
115 120 125
Lys Asp Leu Glu Leu Asn Leu Gln Gln Ile Phe Gly Lys Leu Phe Arg
130 135 140
Lys Tyr Ile Lys Lys Asn Ile Ser Ala Glu Gln Asp Thr Ser Tyr Ser
145 150 155 160
Ser Leu Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr
165 170 175
Ile Trp Leu Ala Met Lys His Gly Thr Thr Cys Ser Ser Gly Ser Gly
180 185 190
Asp Asn Gly Asp Gly Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp
195 200 205
Met Pro Thr Thr Asp Phe Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu
210 215 220
Trp Val Glu His Phe Cys Lys Gln Arg Gln Glu Lys Val Asn Ala Val
225 230 235 240
Ile Thr Asn Cys Lys Ser Cys Lys Glu Ser Gly Gly Thr Cys Asn Ser
245 250 255
Asp Cys Glu Lys Lys Cys Lys Asp Glu Cys Glu Lys Tyr Lys Lys Phe
260 265 270
Ile Glu Glu Cys Arg Thr Ala Ala Asp Gly Thr Ala Gly Ser Ser Trp
275 280 285
Ser Lys Arg Trp Asp Gln Ile Tyr Lys Met Tyr Ser Lys His Ile Glu
290 295 300
Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Thr Ser
305 310 315 320
Ser Thr Thr Asn Ala Ala Glu Asn Lys Cys Val Gln Ser
325 330
<210> 8
<211> 269
<212> PRT
<213> 恶性疟原虫
<400> 8
Asp Tyr Ile Lys Asp Asp Pro Tyr Ser Lys Glu Tyr Thr Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Ser Ser Asp Ala Asn Thr Ser Ser Gly Glu Thr
20 25 30
Ala Asn His Asn Asp Glu Ala Cys Asn Cys Asn Glu Ser Glu Ile Ala
35 40 45
Ser Val Glu Gln Ala Ser Ile Ser Asp Arg Ser Ser Gln Lys Ala Tyr
50 55 60
Ile Thr His Ser Ser Ile Lys Thr Asn Lys Lys Lys Val Cys Lys Tyr
65 70 75 80
Val Lys Leu Gly Ile Asn Asn Asn Asp Lys Val Leu Arg Val Cys Val
85 90 95
Ile Glu Asp Thr Ser Leu Ser Gly Val Glu Asn Cys Cys Phe Lys Asp
100 105 110
Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Arg Gly Ser
115 120 125
Ser Phe Asn Asp Ser Cys Asn Asn Asn Asn Glu Glu Ala Cys Gln Lys
130 135 140
Lys Leu Glu Lys Val Leu Ala Ser Leu Thr Asn Gly Tyr Lys Cys Glu
145 150 155 160
Lys Cys Lys Ser Gly Thr Ser Arg Ser Lys Lys Lys Trp Ile Trp Lys
165 170 175
Lys Ser Ser Gly Lys Glu Gly Gly Leu Gln Lys Glu Tyr Ala Asn Thr
180 185 190
Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu Gly Asn Leu Pro
195 200 205
Lys Leu Glu Asn Val Cys Lys Gly Val Thr Asp Ile Asn Phe Asp Thr
210 215 220
Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Ala Ala Phe His Glu Gly
225 230 235 240
Lys Asn Leu Lys Pro Ser His Gln Asn Lys Asn Asp Asp Asn Asn Ser
245 250 255
Lys Leu Cys Lys Asp Leu Lys Tyr Ser Phe Ala Asp Tyr
260 265
<210> 9
<211> 333
<212> PRT
<213> 恶性疟原虫
<400> 9
Lys Cys Asp Lys Cys Lys Ser Gly Thr Ser Arg Ser Lys Lys Lys Trp
1 5 10 15
Thr Trp Arg Lys Ser Ser Gly Asn Lys Glu Gly Leu Gln Lys Glu Tyr
20 25 30
Ala Asn Thr Ile Gly Leu Pro Pro Arg Thr His Ser Leu Tyr Leu Gly
35 40 45
Asn Leu Arg Lys Leu Glu Asn Val Cys Glu Asp Val Thr Asp Ile Asn
50 55 60
Phe Asp Thr Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Ala Ala Phe
65 70 75 80
His Glu Gly Lys Asn Leu Lys Thr Thr Tyr Pro Gln Asn Lys Asn Asp
85 90 95
Asp Asn Asn Ser Lys Leu Cys Lys Ala Leu Lys Tyr Ser Phe Ala Asp
100 105 110
Tyr Gly Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Asp Phe Thr
115 120 125
Lys Asp Leu Glu Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe Arg
130 135 140
Lys Tyr Ile Lys Lys Asn Ile Ser Thr Glu Gln His Thr Ser Tyr Ser
145 150 155 160
Ser Leu Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr
165 170 175
Ile Trp Leu Ala Met Lys His Gly Ala Glu Met Asn Gly Thr Thr Cys
180 185 190
Ser Cys Ser Gly Asp Ser Ser Asp Asp Ile Pro Thr Ile Asp Leu Ile
195 200 205
Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His Phe Cys Lys
210 215 220
Gln Arg Gln Ala Lys Val Asn Ala Val Ile Asn Ser Cys Asn Ser Cys
225 230 235 240
Lys Asn Thr Ser Gly Glu Arg Lys Leu Gly Gly Thr Cys Gly Ser Glu
245 250 255
Cys Lys Thr Glu Cys Lys Asn Lys Cys Asp Ala Tyr Lys Glu Phe Ile
260 265 270
Asp Gly Thr Gly Ser Gly Gly Gly Thr Gly Thr Ala Gly Ser Ser Trp
275 280 285
Val Lys Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys Tyr Ile Glu
290 295 300
Asp Ala Lys Arg Asn Arg Lys Ala Gly Ser Lys Asn Cys Gly Thr Ser
305 310 315 320
Ser Thr Thr Asn Ala Ala Glu Ser Lys Cys Val Gln Ser
325 330
<210> 10
<211> 650
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 10
Ser Tyr Val Lys Asn Asn Pro Tyr Ser Ala Glu Tyr Val Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Ser Ser Asp Ala Asn Thr Ser Ser Glu Thr Pro
20 25 30
Ser Lys Tyr Tyr Asp Glu Val Cys Asn Cys Asn Glu Ser Glu Ile Ser
35 40 45
Ser Val Gly Gln Ala Gln Thr Ser Gly Pro Ser Ser Asn Lys Thr Cys
50 55 60
Ile Thr His Ser Ser Ile Lys Thr Asn Lys Lys Lys Val Cys Lys Asp
65 70 75 80
Val Lys Leu Gly Ile Asn Asn Asn Asp Lys Val Leu Arg Val Cys Val
85 90 95
Ile Glu Asp Thr Ser Leu Ser Gly Val Asp Asn Cys Cys Cys Gln Asp
100 105 110
Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Lys Asn Gln Ser Gly
115 120 125
Ser Ser Ser Asn Gly Ser Cys Asn Asn Lys Asn Gln Asp Glu Cys Gln
130 135 140
Lys Lys Leu Glu Lys Val Phe Ala Ser Leu Thr Asn Gly Tyr Lys Cys
145 150 155 160
Asp Lys Cys Lys Ser Gly Thr Ser Arg Ser Lys Lys Lys Trp Ile Trp
165 170 175
Arg Lys Ser Ser Gly Asn Glu Glu Gly Leu Gln Lys Glu Tyr Ala Asn
180 185 190
Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu Gly Asn Leu
195 200 205
Arg Lys Leu Glu Asn Val Cys Lys Gly Val Thr Asp Ile Asn Phe Asp
210 215 220
Thr Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Ala Ala Phe His Glu
225 230 235 240
Gly Lys Asn Leu Lys Thr Thr Tyr Pro Gln Asn Lys Lys Lys Leu Cys
245 250 255
Lys Asp Leu Lys Tyr Ser Phe Ala Asp Tyr Gly Asp Leu Ile Lys Gly
260 265 270
Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp Leu Glu Leu Asn Leu
275 280 285
Gln Lys Ala Phe Gly Lys Leu Phe Arg Lys Tyr Ile Lys Lys Asn Ile
290 295 300
Ser Thr Glu Gln His Thr Leu Tyr Ser Ser Leu Asp Glu Leu Arg Glu
305 310 315 320
Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Leu Ala Met Lys His
325 330 335
Gly Ala Gly Met Asn Ser Thr Thr Cys Cys Gly Asp Gly Ser Val Thr
340 345 350
Gly Ser Gly Ser Ser Cys Asp Asp Ile Pro Thr Ile Asp Leu Ile Pro
355 360 365
Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His Phe Cys Lys Gln
370 375 380
Arg Gln Glu Lys Val Asn Ala Val Ile Glu Asn Cys Asn Ser Cys Lys
385 390 395 400
Glu Cys Gly Asp Thr Cys Asn Gly Glu Cys Lys Thr Glu Cys Glu Lys
405 410 415
Lys Cys Lys Ile Glu Cys Glu Lys Tyr Lys Thr Phe Ile Glu Glu Cys
420 425 430
Val Thr Ala Val Gly Gly Thr Ser Gly Ser Pro Trp Ser Lys Arg Trp
435 440 445
Asp Gln Ile Tyr Lys Arg Tyr Ser Lys Tyr Ile Glu Asp Ala Lys Arg
450 455 460
Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Ile Thr Thr Gly Thr Ile
465 470 475 480
Ser Gly Glu Ser Ser Gly Ala Asn Ser Gly Val Thr Thr Thr Glu Asn
485 490 495
Lys Cys Val Gln Ser Asp Ile Asp Ser Phe Phe Lys His Leu Ile Asp
500 505 510
Ile Gly Leu Thr Thr Pro Ser Ser Tyr Leu Ser Ile Val Leu Asp Asp
515 520 525
Asn Ile Cys Gly Ala Asp Asn Ala Pro Trp Thr Thr Tyr Thr Thr Tyr
530 535 540
Thr Thr Tyr Thr Thr Thr Lys Asn Cys Asp Ile Lys Lys Lys Thr Pro
545 550 555 560
Lys Ser Gln Pro Ile Asn Thr Ser Val Val Val Asn Val Pro Ser Pro
565 570 575
Leu Gly Asn Thr Pro His Gly Tyr Lys Tyr Ala Cys Gln Cys Lys Ile
580 585 590
Pro Thr Thr Glu Glu Ser Cys Asp Asp Arg Lys Glu Tyr Met Asn Gln
595 600 605
Trp Ile Ile Asp Thr Ser Lys Lys Gln Lys Gly Ser Gly Ser Thr Asn
610 615 620
Asn Asp Tyr Glu Leu Tyr Thr Tyr Asn Gly Val Lys Glu Thr Lys Leu
625 630 635 640
Pro Lys Lys Ser Ser Ser Ser Lys Leu Asp
645 650
<210> 11
<211> 643
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 11
Ser Tyr Val Lys Asp Asp Pro Tyr Ser Ala Glu Tyr Val Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Ser Ser Asp Ala Asn Thr Ser Ser Glu Thr Pro
20 25 30
Ser Lys Tyr Tyr Asp Glu Val Cys Asn Cys Asn Glu Ser Glu Ile Ser
35 40 45
Ser Val Gly Gln Ala Gln Thr Ser Gly Pro Ser Ser Asn Lys Thr Cys
50 55 60
Ile Thr His Ser Ser Ile Lys Thr Asn Lys Lys Lys Val Cys Lys Asp
65 70 75 80
Val Lys Leu Gly Ile Asn Asn Asn Asp Lys Val Leu Arg Val Cys Val
85 90 95
Ile Glu Asp Thr Ser Leu Ser Gly Val Asp Asn Cys Cys Cys Gln Asp
100 105 110
Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Lys Asn Gln Ser Gly
115 120 125
Ser Ser Ser Asn Gly Ser Cys Asn Asn Lys Asn Gln Asp Glu Cys Gln
130 135 140
Lys Lys Leu Glu Lys Val Phe Ala Ser Leu Thr Asn Gly Tyr Lys Cys
145 150 155 160
Asp Lys Cys Lys Ser Gly Thr Ser Arg Ser Lys Lys Lys Trp Ile Trp
165 170 175
Arg Lys Ser Ser Gly Asn Glu Glu Gly Leu Gln Lys Glu Tyr Ala Asn
180 185 190
Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu Gly Asn Leu
195 200 205
Pro Lys Leu Glu Asn Val Cys Lys Gly Val Thr Asp Ile Ile Tyr Asp
210 215 220
Thr Lys Glu Lys Phe Leu Ser Gly Cys Leu Ile Ala Ala Phe His Glu
225 230 235 240
Gly Lys Asn Leu Lys Thr Ser His Glu Lys Lys Asn Asp Asp Asn Gly
245 250 255
Lys Lys Leu Cys Lys Ala Leu Glu Tyr Ser Phe Ala Asp Tyr Gly Asp
260 265 270
Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Asp Phe Thr Lys Asp Leu
275 280 285
Glu Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe Arg Lys Tyr Ile
290 295 300
Lys Lys Asn Asn Thr Ala Glu Gln Asp Thr Ser Tyr Ser Ser Leu Asp
305 310 315 320
Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Thr
325 330 335
Ala Met Lys His Gly Ala Gly Met Asn Ser Thr Thr Cys Ser Gly Asp
340 345 350
Gly Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp Met Pro Thr Ile
355 360 365
Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His
370 375 380
Phe Cys Lys Gln Arg Gln Glu Lys Val Lys Asp Val Ile Thr Asn Cys
385 390 395 400
Asn Ser Cys Lys Glu Cys Gly Asp Thr Cys Asn Gly Glu Cys Lys Thr
405 410 415
Glu Cys Lys Thr Lys Cys Lys Gly Glu Cys Glu Lys Tyr Lys Asn Phe
420 425 430
Ile Glu Glu Cys Asn Gly Thr Ala Asp Gly Gly Thr Ser Gly Ser Ser
435 440 445
Trp Ser Lys Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys Tyr Ile
450 455 460
Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Thr
465 470 475 480
Ser Ser Thr Thr Asn Ala Ala Ala Ser Thr Thr Glu Asn Lys Cys Val
485 490 495
Gln Ser Asp Ile Asp Ser Phe Phe Lys His Leu Ile Asp Ile Gly Leu
500 505 510
Thr Thr Pro Ser Ser Tyr Leu Ser Asn Val Leu Asp Asp Asn Ile Cys
515 520 525
Gly Glu Asp Lys Ala Pro Trp Thr Thr Tyr Thr Thr Tyr Thr Thr Lys
530 535 540
Asn Cys Asp Ile Gln Lys Lys Thr Pro Lys Pro Gln Ser Cys Asp Thr
545 550 555 560
Leu Val Val Val Asn Val Pro Ser Pro Leu Gly Asn Thr Pro His Gly
565 570 575
Tyr Lys Tyr Val Cys Glu Cys Lys Ile Pro Thr Thr Glu Glu Thr Cys
580 585 590
Asp Asp Arg Lys Glu Tyr Met Asn Gln Trp Ile Ile Asp Thr Ser Lys
595 600 605
Lys Gln Lys Gly Ser Gly Ser Thr Asn Asn Asp Tyr Glu Leu Tyr Thr
610 615 620
Tyr Asn Gly Val Gln Ile Lys Gln Ala Ala Gly Thr Leu Lys Asn Ser
625 630 635 640
Lys Leu Asp
<210> 12
<211> 269
<212> PRT
<213> 恶性疟原虫
<400> 12
Asn Tyr Ile Lys Gly Asp Pro Tyr Ser Ala Glu Tyr Ala Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Ser Ser Asp Thr Glu Asn Ala Ser Glu Lys Ile
20 25 30
Gln Lys Asn Asn Asp Glu Val Cys Asn Cys Asn Glu Ser Glu Ile Ala
35 40 45
Ser Val Glu Gln Ala Pro Ile Ser Asp Arg Ser Ser Gln Lys Ala Cys
50 55 60
Ile Thr His Ser Ser Ile Lys Ala Asn Lys Lys Lys Val Cys Lys His
65 70 75 80
Val Lys Leu Gly Val Arg Glu Asn Asp Lys Asp Leu Lys Ile Cys Val
85 90 95
Ile Glu Asp Thr Ser Leu Ser Gly Val Asp Asn Cys Cys Cys Gln Asp
100 105 110
Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Ser Gly Ser
115 120 125
Ser Ser Asn Gly Ser Cys Asn Asn Asn Asn Glu Glu Ile Cys Gln Lys
130 135 140
Lys Leu Glu Lys Val Leu Ala Ser Leu Thr Asn Gly Tyr Lys Cys Asp
145 150 155 160
Lys Cys Lys Ser Gly Thr Ser Thr Val Asn Lys Asn Trp Ile Trp Lys
165 170 175
Lys Tyr Ser Gly Lys Glu Gly Gly Leu Gln Glu Glu Tyr Ala Asn Thr
180 185 190
Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu Gly Asn Leu Pro
195 200 205
Lys Leu Glu Asn Val Cys Glu Asp Val Lys Asp Ile Asn Phe Asp Thr
210 215 220
Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Ala Ala Phe His Glu Gly
225 230 235 240
Lys Asn Leu Lys Thr Ser Asn Lys Lys Lys Asn Asp Asp Asn Asn Ser
245 250 255
Lys Leu Cys Lys Ala Leu Lys Tyr Ser Phe Ala Asp Tyr
260 265
<210> 13
<211> 344
<212> PRT
<213> 恶性疟原虫
<400> 13
Lys Cys Asp Lys Cys Lys Ser Gly Thr Ser Thr Val Asn Lys Lys Trp
1 5 10 15
Ile Trp Lys Lys Tyr Ser Gly Thr Glu Gly Gly Leu Gln Glu Glu Tyr
20 25 30
Ala Asn Thr Ile Ala Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu Gly
35 40 45
Asn Leu Pro Lys Leu Glu Asn Val Cys Lys Asp Val Thr Asp Ile Asn
50 55 60
Phe Asp Thr Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Ala Ala Phe
65 70 75 80
His Glu Gly Lys Asn Leu Lys Thr Thr Tyr Leu Glu Lys Lys Lys Gly
85 90 95
Asp Asn Gly Lys Lys Asn Asp Asp Asn Asn Ser Lys Leu Cys Lys Ala
100 105 110
Leu Lys Tyr Ser Phe Ala Asp Tyr Gly Asp Leu Ile Lys Gly Thr Ser
115 120 125
Ile Trp Asp Asn Asp Phe Thr Lys Asp Leu Glu Leu Asn Leu Gln Gln
130 135 140
Ile Phe Gly Lys Leu Phe Arg Lys Tyr Ile Lys Lys Asn Ile Ala Ser
145 150 155 160
Asp Glu Asn Thr Leu Tyr Ser Ser Leu Asp Glu Leu Arg Glu Ser Trp
165 170 175
Trp Asn Thr Asn Lys Lys Tyr Ile Trp Leu Ala Met Lys His Gly Ala
180 185 190
Gly Met Asn Ser Thr Met Cys Asn Ala Asp Gly Ser Val Thr Gly Ser
195 200 205
Gly Ser Ser Cys Asp Asp Ile Pro Thr Ile Asp Leu Ile Pro Gln Tyr
210 215 220
Leu Arg Phe Leu Gln Glu Trp Val Glu His Phe Cys Lys Gln Arg Gln
225 230 235 240
Ala Lys Val Lys Asp Val Ile Thr Asn Cys Asn Ser Cys Lys Glu Cys
245 250 255
Gly Gly Thr Cys Asn Gly Glu Cys Lys Thr Glu Cys Glu Lys Lys Cys
260 265 270
Lys Gly Glu Cys Asp Ala Tyr Lys Lys Phe Ile Glu Glu Cys Lys Gly
275 280 285
Lys Ala Asp Glu Gly Thr Ser Gly Ser Ser Trp Ser Lys Arg Trp Asp
290 295 300
Gln Ile Tyr Lys Arg Tyr Ser Lys Tyr Ile Glu Asp Ala Lys Arg Asn
305 310 315 320
Arg Lys Ala Gly Thr Lys Asn Cys Gly Pro Ser Ser Thr Thr Ser Thr
325 330 335
Ala Glu Ser Lys Cys Val Gln Ser
340
<210> 14
<211> 334
<212> PRT
<213> 恶性疟原虫
<400> 14
Lys Cys Asp Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Asn Ile
1 5 10 15
Trp Ile Trp Lys Lys Ser Ser Gly Thr Glu Gly Gly Leu Gln Lys Glu
20 25 30
Tyr Ala Asn Thr Ile Ala Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu
35 40 45
Gly Asn Leu Arg Lys Leu Glu Asn Val Cys Glu Asp Val Lys Asp Ile
50 55 60
Asn Phe Asp Thr Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Ala Ala
65 70 75 80
Phe His Glu Gly Lys Asn Leu Lys Lys Arg Tyr Leu Glu Lys Lys Asn
85 90 95
Gly Asp Asn Asn Ser Lys Leu Cys Lys Ala Leu Lys Tyr Ser Phe Ala
100 105 110
Asp Tyr Gly Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr
115 120 125
Thr Lys Asp Leu Glu Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe
130 135 140
Arg Lys Tyr Ile Lys Lys Asn Asn Thr Ala Glu Gln His Thr Ser Tyr
145 150 155 160
Ser Ser Leu Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys
165 170 175
Tyr Ile Trp Leu Ala Met Lys His Gly Thr Thr Cys Ser Ser Gly Ser
180 185 190
Gly Asp Asn Gly Ser Ile Ser Cys Asp Asp Ile Pro Thr Ile Asp Leu
195 200 205
Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His Phe Cys
210 215 220
Glu Gln Arg Gln Gly Lys Val Asn Ala Val Ile Glu Asn Cys Asn Ser
225 230 235 240
Cys Lys Asn Thr Ser Ser Lys Thr Lys Leu Gly Gly Thr Cys Asn Gly
245 250 255
Glu Cys Lys Thr Glu Cys Lys Gly Glu Cys Asp Ala Tyr Lys Glu Phe
260 265 270
Ile Glu Lys Cys Lys Gly Thr Ala Ala Glu Gly Thr Ser Gly Ser Ser
275 280 285
Trp Val Lys Arg Trp Tyr Gln Ile Tyr Met Arg Tyr Ser Lys Tyr Ile
290 295 300
Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Thr
305 310 315 320
Ser Ser Thr Thr Ser Thr Ala Glu Ser Lys Cys Val Gln Ser
325 330
<210> 15
<211> 332
<212> PRT
<213> 恶性疟原虫
<400> 15
Lys Cys Asp Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Asn Ile
1 5 10 15
Trp Ile Trp Lys Lys Ser Ser Gly Thr Glu Gly Gly Leu Gln Lys Glu
20 25 30
Tyr Ala Asn Thr Ile Ala Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu
35 40 45
Gly Asn Leu Arg Lys Leu Glu Asn Val Cys Glu Asp Val Lys Asp Ile
50 55 60
Asn Phe Asp Thr Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Ala Ala
65 70 75 80
Phe His Glu Gly Lys Asn Leu Lys Lys Arg Tyr Leu Glu Lys Lys Asn
85 90 95
Gly Asp Asn Asn Ser Lys Leu Cys Lys Ala Leu Lys Tyr Ser Phe Ala
100 105 110
Asp Tyr Gly Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr
115 120 125
Thr Lys Asp Leu Glu Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe
130 135 140
Arg Lys Tyr Ile Lys Lys Asn Asn Thr Ala Glu Gln Asp Thr Ser Tyr
145 150 155 160
Ser Ser Leu Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys
165 170 175
Tyr Ile Trp Thr Ala Met Lys His Gly Thr Thr Cys Ser Ser Gly Ser
180 185 190
Gly Asp Asn Gly Ser Ile Ser Cys Asp Asp Ile Pro Thr Ile Asp Leu
195 200 205
Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His Phe Cys
210 215 220
Glu Gln Arg Gln Glu Lys Val Lys Asp Val Ile Lys Asn Cys Asn Ser
225 230 235 240
Cys Lys Glu Cys Gly Gly Thr Cys Asn Gly Glu Cys Lys Thr Glu Cys
245 250 255
Lys Asn Lys Cys Lys Asp Glu Cys Asp Ala Tyr Lys Lys Phe Ile Glu
260 265 270
Glu Cys Glu Gly Lys Ala Ala Glu Gly Thr Ser Gly Ser Ser Trp Ser
275 280 285
Lys Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys Tyr Ile Glu Asp
290 295 300
Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Thr Ser Ser
305 310 315 320
Thr Thr Ser Thr Ala Glu Asn Lys Cys Val Gln Ser
325 330
<210> 16
<211> 267
<212> PRT
<213> 恶性疟原虫
<400> 16
Asn Tyr Ile Lys Asp Asp Pro Tyr Ser Ala Glu Tyr Thr Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Ser Ser Asp Thr Glu Asn Ala Ser Glu Lys Ile
20 25 30
Gln Lys Asn Asn Asp Glu Val Cys Asn Pro Asn Glu Ser Gly Ile Ala
35 40 45
Cys Val Glu Leu Ala Gln Thr Ser Gly Ser Ser Ser Asn Lys Thr Cys
50 55 60
Asn Thr His Ser Phe Ile Lys Ala Asn Lys Lys Lys Val Cys Lys Asp
65 70 75 80
Val Lys Leu Gly Ile Asn Lys Lys Asp Lys Asp Leu Lys Ile Cys Val
85 90 95
Ile Glu Asp Asp Ser Leu Arg Gly Val Asp Asn Cys Cys Cys Gln Asp
100 105 110
Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Lys Asn Gln Ser Gly
115 120 125
Ser Ser Ser Asn Gly Ser Cys Asn Asn Lys Asn Gln Glu Ala Cys Gln
130 135 140
Lys Lys Leu Glu Asn Val Phe Ala Ser Leu Thr Asn Gly Tyr Lys Cys
145 150 155 160
Glu Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Lys Asn Trp Ile
165 170 175
Trp Lys Lys Tyr Ser Val Lys Glu Glu Gly Leu Gln Lys Glu Tyr Ala
180 185 190
Asn Thr Ile Ala Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu Gly Asn
195 200 205
Leu Pro Lys Leu Gly Asn Val Cys Lys Gly Val Thr Asp Ile Asn Phe
210 215 220
Asp Thr Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Ala Ala Phe His
225 230 235 240
Glu Gly Lys Asn Leu Lys Thr Thr Tyr Leu Gln Asn Lys Lys Lys Leu
245 250 255
Cys Lys Ala Leu Lys Tyr Ser Phe Ala Asp Tyr
260 265
<210> 17
<211> 263
<212> PRT
<213> 恶性疟原虫
<400> 17
Asp Tyr Ile Lys Gly Asp Pro Tyr Phe Ala Glu Tyr Ala Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Ser Ser Asp Ala Asn Thr Ser Ser Gly Glu Thr
20 25 30
Ala Asn His Asn Asp Glu Ala Cys Asn Pro Asn Glu Ser Glu Ile Ala
35 40 45
Ser Val Glu Gln Ala Ser Ile Ser Asp Arg Ser Ser Gln Lys Ala Cys
50 55 60
Asn Thr His Ser Ser Ile Lys Ala Asn Lys Lys Lys Glu Cys Lys His
65 70 75 80
Val Lys Leu Gly Val Arg Glu Asn Asp Lys Asp Leu Lys Ile Cys Val
85 90 95
Ile Glu Asp Thr Ser Leu Ser Gly Val Asp Asn Cys Cys Cys Gln Asp
100 105 110
Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Arg Gly Ser
115 120 125
Ser Ser Asn Gly Ser Cys Asp Lys Asn Ser Glu Glu Ile Cys Gln Lys
130 135 140
Lys Leu Asp Glu Ala Leu Ala Ser Leu His Asn Gly Tyr Lys Asn Gln
145 150 155 160
Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Lys Asn Lys Trp Ile Trp
165 170 175
Lys Lys Ser Ser Gly Asn Glu Lys Gly Leu Gln Lys Glu Tyr Ala Asn
180 185 190
Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu Gly Asn Leu
195 200 205
Pro Lys Leu Glu Asn Val Cys Glu Asp Val Thr Asp Ile Asn Phe Asp
210 215 220
Thr Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Ala Ala Phe His Glu
225 230 235 240
Gly Lys Asn Leu Lys Thr Thr Tyr Pro Gln Asn Lys Asn Asp Asp Asn
245 250 255
Gly Lys Lys Leu Cys Lys Asp
260
<210> 18
<211> 338
<212> PRT
<213> 恶性疟原虫
<400> 18
Lys Cys Asp Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Asn Ile
1 5 10 15
Trp Ile Trp Lys Lys Ser Ser Gly Asn Lys Lys Gly Leu Gln Lys Glu
20 25 30
Tyr Ala Asn Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu
35 40 45
Gly Asn Leu Pro Lys Leu Glu Asn Val Cys Lys Asp Val Thr Asp Ile
50 55 60
Asn Phe Asp Thr Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Ala Ala
65 70 75 80
Phe His Glu Gly Lys Asn Leu Lys Ile Ser Asn Glu Lys Lys Asn Asp
85 90 95
Asp Asn Gly Lys Lys Leu Cys Lys Asp Leu Lys Tyr Ser Phe Ala Asp
100 105 110
Tyr Gly Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr
115 120 125
Lys Asp Leu Glu Leu Asn Leu Gln Asn Asn Phe Gly Lys Leu Phe Arg
130 135 140
Lys Tyr Ile Lys Lys Asn Asn Thr Ala Glu Gln His Thr Leu Tyr Ser
145 150 155 160
Ser Leu Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr
165 170 175
Ile Trp Leu Ala Met Lys His Gly Thr Thr Cys Ser Ser Gly Ser Gly
180 185 190
Asp Asn Gly Asp Gly Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp
195 200 205
Met Ser Thr Ile Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu
210 215 220
Trp Val Glu His Phe Cys Lys Gln Arg Gln Glu Lys Val Asn Ala Val
225 230 235 240
Ile Glu Asn Cys Asn Ser Cys Lys Asn Thr Ser Ser Lys Thr Lys Leu
245 250 255
Gly Gly Thr Cys Asn Gly Glu Cys Lys Thr Glu Cys Glu Lys Lys Cys
260 265 270
Lys Asp Glu Cys Glu Lys Tyr Lys Glu Phe Ile Glu Glu Cys Lys Arg
275 280 285
Gly Asp Gly Thr Ala Gly Ser Pro Trp Val Lys Arg Trp Asp Gln Ile
290 295 300
Tyr Met Arg Tyr Ser Lys Tyr Ile Glu Asp Ala Lys Arg Asn Arg Lys
305 310 315 320
Ala Gly Thr Lys Ser Cys Gly Thr Ser Ala Ala Glu Asn Lys Cys Val
325 330 335
Gln Ser
<210> 19
<211> 341
<212> PRT
<213> 恶性疟原虫
<400> 19
Lys Cys Asp Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Asn Ile
1 5 10 15
Trp Ile Trp Lys Lys Ser Ser Gly Asp Glu Lys Gly Leu Gln Lys Glu
20 25 30
Tyr Ala Asn Thr Ile Ala Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu
35 40 45
Gly Asn Leu Pro Lys Leu Glu Asn Val Cys Lys Asp Val Thr Asp Ile
50 55 60
Asn Phe Asp Thr Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Ala Ala
65 70 75 80
Phe His Glu Gly Lys Asn Leu Lys Thr Ser His Gln Asn Lys Asn Ala
85 90 95
Asp Asn Gly Lys Lys Asn Asp Asp Asn Gly Lys Lys Leu Cys Lys Ala
100 105 110
Leu Lys Tyr Ser Phe Ala Asp Tyr Gly Asp Leu Ile Lys Gly Thr Ser
115 120 125
Ile Trp Asp Asn Glu Tyr Thr Lys Asp Leu Glu Leu Asn Leu Gln Gln
130 135 140
Ile Phe Gly Lys Leu Phe Arg Lys Tyr Ile Lys Arg Asn Asn Thr Ala
145 150 155 160
Glu Gln His Thr Leu Tyr Ser Ser Leu Asp Glu Leu Arg Glu Ser Trp
165 170 175
Trp Asn Thr Asn Lys Lys Tyr Ile Trp Leu Ala Met Lys His Gly Thr
180 185 190
Thr Cys Ser Ser Gly Ser Gly Asp Asn Gly Asp Gly Ser Val Thr Gly
195 200 205
Ser Gly Ser Ser Cys Asp Asp Met Ser Thr Ile Asp Leu Ile Pro Gln
210 215 220
Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His Phe Cys Lys Gln Arg
225 230 235 240
Gln Glu Lys Val Lys Asp Val Ile Thr Asn Cys Asn Ser Cys Lys Glu
245 250 255
Cys Gly Gly Thr Cys Gly Ser Asp Cys Lys Thr Lys Cys Glu Ala Tyr
260 265 270
Lys Lys Phe Ile Glu Glu Cys Asn Gly Thr Ala Asp Gly Gly Thr Ser
275 280 285
Gly Ser Ser Trp Ser Lys Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser
290 295 300
Lys Tyr Ile Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn
305 310 315 320
Cys Gly Pro Ser Ser Gly Ala Asn Ser Gly Val Thr Thr Thr Glu Asn
325 330 335
Lys Cys Val Gln Ser
340
<210> 20
<211> 352
<212> PRT
<213> 恶性疟原虫
<400> 20
Lys Cys Glu Lys Cys Glu Ser Glu Gln Ser Lys Lys Asn Asn Lys Tyr
1 5 10 15
Trp Ile Trp Lys Lys Ser Ser Gly Asn Gly Glu Gly Leu Gln Glu Glu
20 25 30
Tyr Ala Asn Thr Ile Ala Leu Pro Pro Arg Thr His Ser Leu Cys Leu
35 40 45
Val Cys Leu His Glu Lys Glu Gly Lys Lys Thr Gln Glu Leu Lys Asn
50 55 60
Ile Arg Thr Asn Ser Glu Leu Leu Lys Glu Arg Ile Ile Ala Ala Phe
65 70 75 80
His Glu Gly Lys Asn Leu Lys Thr Ser Pro Gln Asn Lys Asn Asp Asn
85 90 95
Gly Lys Lys Leu Cys Lys Asp Leu Lys Tyr Ser Phe Ala Asp Tyr Gly
100 105 110
Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp
115 120 125
Leu Glu Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe Arg Lys Tyr
130 135 140
Ile Lys Lys Asn Asn Thr Ala Glu Gln His Thr Leu Tyr Ser Ser Leu
145 150 155 160
Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp
165 170 175
Leu Ala Met Lys His Gly Ala Gly Met Asn Ser Thr Met Cys Asn Ala
180 185 190
Asp Gly Ser Val Thr Gly Ser Ser Asp Ser Gly Ser Thr Thr Cys Cys
195 200 205
Gly Asp Asn Gly Ser Ile Ser Cys Asp Asp Met Pro Thr Ile Asp Leu
210 215 220
Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His Phe Cys
225 230 235 240
Glu Gln Arg Gln Glu Lys Val Asn Ala Val Ile Thr Asn Cys Lys Ser
245 250 255
Cys Lys Glu Cys Gly Gly Thr Cys Asn Ser Asp Cys Glu Lys Lys Cys
260 265 270
Lys Ala Tyr Lys Glu Phe Ile Glu Lys Cys Lys Gly Gly Gly Thr Glu
275 280 285
Gly Thr Ser Gly Ser Ser Trp Ser Lys Arg Trp Asp Gln Ile Tyr Lys
290 295 300
Arg His Ser Lys His Ile Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly
305 310 315 320
Thr Lys Asn Cys Gly Ile Thr Thr Gly Thr Ile Ser Gly Glu Ser Ser
325 330 335
Gly Ala Asn Ser Gly Val Thr Thr Thr Glu Asn Lys Cys Val Gln Ser
340 345 350
<210> 21
<211> 344
<212> PRT
<213> 恶性疟原虫
<400> 21
Lys Cys Asp Lys Cys Lys Ser Gly Thr Ser Arg Ser Arg Lys Ile Trp
1 5 10 15
Thr Trp Arg Lys Phe Arg Gly Asn Gly Glu Gly Leu Gln Lys Glu Tyr
20 25 30
Ala Asn Thr Ile Gly Leu Ser Pro Arg Thr Gln Leu Leu Tyr Leu Val
35 40 45
Cys Leu His Glu Lys Gly Lys Lys Thr Gln Glu Leu Lys Asn Ile Ser
50 55 60
Thr Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu
65 70 75 80
Gly Lys Asn Leu Lys Thr Thr Tyr Pro Gln Lys Lys Asn Asp Asp Asn
85 90 95
Gly Lys Lys Leu Cys Lys Ala Leu Lys Tyr Ser Phe Ala Asp Tyr Gly
100 105 110
Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Asp Phe Thr Lys Asp
115 120 125
Leu Glu Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe Arg Lys Tyr
130 135 140
Ile Lys Lys Asn Ile Ala Ser Asp Glu Asn Thr Ser Tyr Ser Ser Leu
145 150 155 160
Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp
165 170 175
Thr Ala Met Lys His Gly Ala Gly Met Asn Gly Thr Thr Cys Cys Gly
180 185 190
Asp Gly Ser Val Thr Gly Ser Ser Asp Ser Gly Ser Thr Thr Cys Cys
195 200 205
Gly Asp Gly Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp Ile Pro
210 215 220
Thr Ile Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val
225 230 235 240
Glu His Phe Cys Glu Gln Arg Gln Glu Lys Val Lys Asp Val Ile Thr
245 250 255
Asn Cys Lys Ser Cys Lys Glu Ser Glu Lys Lys Cys Lys Asn Lys Cys
260 265 270
Asp Ala Tyr Lys Glu Phe Ile Asp Gly Thr Gly Ser Gly Gly Gly Thr
275 280 285
Gly Thr Ala Gly Ser Ser Trp Ser Lys Arg Trp Asp Gln Ile Tyr Met
290 295 300
Arg Tyr Ser Lys Tyr Ile Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly
305 310 315 320
Thr Lys Asn Cys Gly Thr Ser Ser Gly Ala Asn Ser Gly Val Thr Thr
325 330 335
Thr Glu Asn Lys Cys Val Gln Ser
340
<210> 22
<211> 350
<212> PRT
<213> 恶性疟原虫
<400> 22
Lys Cys Glu Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Lys Ile
1 5 10 15
Trp Thr Trp Arg Lys Phe Pro Gly Asn Gly Glu Gly Leu Gln Lys Glu
20 25 30
Tyr Ala Asn Thr Ile Gly Leu Ser Pro Arg Thr Gln Leu Leu Tyr Leu
35 40 45
Val Cys Leu His Glu Lys Gly Lys Lys Thr Gln His Lys Thr Ile Ser
50 55 60
Thr Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu
65 70 75 80
Gly Lys Asn Leu Lys Lys Arg Tyr Leu Glu Lys Lys Lys Gly Asp Asn
85 90 95
Asn Ser Lys Leu Cys Lys Asp Leu Lys Tyr Ser Phe Ala Asp Tyr Gly
100 105 110
Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Asp Phe Thr Lys Asp
115 120 125
Leu Glu Leu Asn Leu Gln Gln Ile Phe Gly Lys Leu Phe Arg Lys Tyr
130 135 140
Ile Lys Lys Asn Ile Ala Ser Asp Glu Asn Thr Ser Tyr Ser Ser Leu
145 150 155 160
Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp
165 170 175
Thr Ala Met Lys His Gly Ala Gly Met Asn Ser Thr Met Cys Asn Gly
180 185 190
Asp Gly Ser Val Thr Gly Ser Ser Asp Ser Gly Ser Thr Thr Cys Ser
195 200 205
Gly Asp Asn Gly Ser Ile Ser Cys Asp Asp Ile Pro Thr Ile Asp Leu
210 215 220
Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His Phe Cys
225 230 235 240
Glu Gln Arg Gln Glu Lys Val Lys Asp Val Ile Lys Asn Cys Asn Ser
245 250 255
Cys Lys Glu Cys Gly Gly Thr Cys Asn Gly Glu Cys Lys Thr Glu Cys
260 265 270
Lys Asn Lys Cys Lys Asp Glu Cys Glu Lys Tyr Lys Asn Phe Ile Glu
275 280 285
Val Cys Thr Gly Gly Asp Gly Thr Ala Gly Ser Pro Trp Ser Lys Arg
290 295 300
Trp Tyr Gln Ile Tyr Met Arg Tyr Ser Lys Tyr Ile Glu Asp Ala Lys
305 310 315 320
Arg Asn Arg Lys Ala Gly Thr Lys Ser Cys Gly Thr Ser Ser Gly Ala
325 330 335
Asn Ser Gly Val Thr Thr Thr Glu Ser Lys Cys Val Gln Ser
340 345 350
<210> 23
<211> 359
<212> PRT
<213> 恶性疟原虫
<400> 23
Lys Cys Glu Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Lys Asn
1 5 10 15
Trp Ile Trp Arg Lys Phe Pro Gly Asn Gly Glu Gly Leu Gln Lys Glu
20 25 30
Tyr Ala Asn Thr Ile Gly Leu Pro Pro Arg Thr His Ser Leu Tyr Leu
35 40 45
Val Cys Leu His Glu Lys Gly Lys Lys Thr Gln Glu Leu Lys Asn Ile
50 55 60
Arg Thr Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His
65 70 75 80
Glu Gly Lys Asn Leu Lys Lys Arg Tyr His Gln Asn Asn Asn Ser Gly
85 90 95
Asn Lys Lys Lys Leu Cys Lys Ala Leu Glu Tyr Ser Phe Ala Asp Tyr
100 105 110
Gly Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys
115 120 125
Asp Leu Glu Leu Asn Leu Gln Gln Ile Phe Gly Lys Leu Phe Arg Lys
130 135 140
Tyr Ile Lys Lys Asn Ile Ser Thr Glu Gln Asp Thr Leu Tyr Ser Ser
145 150 155 160
Leu Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile
165 170 175
Trp Leu Ala Met Lys His Gly Ala Gly Met Asn Ser Thr Thr Cys Cys
180 185 190
Gly Asp Gly Ser Val Thr Gly Ser Ser Asp Ser Gly Ser Thr Thr Cys
195 200 205
Ser Gly Asp Asn Gly Ser Ile Ser Cys Asp Asp Met Pro Thr Ile Asp
210 215 220
Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His Phe
225 230 235 240
Cys Glu Gln Arg Gln Glu Lys Val Lys Asp Val Ile Glu Asn Cys Lys
245 250 255
Ser Cys Lys Asn Thr Ser Gly Glu Arg Ile Ile Gly Gly Thr Cys Asn
260 265 270
Gly Glu Cys Lys Thr Glu Cys Glu Lys Lys Cys Lys Ala Ala Cys Glu
275 280 285
Ala Tyr Lys Thr Phe Ile Glu Glu Cys Glu Gly Lys Ala Ala Glu Gly
290 295 300
Thr Ser Gly Ser Ser Trp Ser Lys Arg Trp Tyr Gln Ile Tyr Met Arg
305 310 315 320
Tyr Ser Lys Tyr Ile Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr
325 330 335
Lys Asn Cys Gly Lys Ser Ser Gly Ala Asn Ser Gly Val Thr Thr Thr
340 345 350
Glu Asn Lys Cys Val Gln Ser
355
<210> 24
<211> 270
<212> PRT
<213> 恶性疟原虫
<400> 24
Asn Tyr Ile Lys Asp Asp Pro Tyr Ser Lys Glu Tyr Val Thr Lys Leu
1 5 10 15
Ser Phe Ile Pro Asn Ser Ser Asp Ala Asn Asn Pro Ser Gly Glu Thr
20 25 30
Ala Asn His Asn Asp Glu Val Cys Asn Pro Asn Glu Ser Glu Ile Ser
35 40 45
Ser Val Glu His Ala Gln Thr Ser Val Leu Leu Ser Gln Lys Ala Tyr
50 55 60
Ile Thr His Ser Ser Ile Lys Ala Asn Lys Lys Lys Val Cys Lys Tyr
65 70 75 80
Val Lys Leu Gly Val Arg Glu Asn Asp Lys Asp Leu Lys Ile Cys Val
85 90 95
Ile Glu Asp Asp Ser Leu Arg Gly Val Glu Asn Cys Cys Phe Lys Asp
100 105 110
Phe Leu Arg Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Arg Glu Ser
115 120 125
Ser Ser Asn Gly Ser Cys Asn Asn Asn Asn Glu Glu Ala Cys Glu Lys
130 135 140
Asn Leu Asp Glu Ala Leu Ala Ser Leu Thr Asn Cys Tyr Lys Asn Gln
145 150 155 160
Lys Cys Lys Ser Gly Thr Ser Thr Val Asn Asn Asn Lys Trp Ile Trp
165 170 175
Lys Lys Ser Ser Gly Lys Glu Gly Gly Leu Gln Lys Glu Tyr Ala Asn
180 185 190
Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Cys Leu Val Val Cys
195 200 205
Leu Asp Glu Lys Glu Gly Lys Thr Gln Glu Leu Lys Asn Ile Arg Thr
210 215 220
Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu Gly
225 230 235 240
Lys Asn Leu Lys Lys Arg Tyr His Gln Asn Lys Asn Asp Asp Asn Asn
245 250 255
Ser Lys Leu Cys Lys Ala Leu Lys Tyr Ser Phe Ala Asp Tyr
260 265 270
<210> 25
<211> 334
<212> PRT
<213> 恶性疟原虫
<400> 25
Lys Cys Asp Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Lys Tyr
1 5 10 15
Trp Ile Trp Lys Lys Tyr Ser Val Lys Glu Gly Gly Leu Gln Lys Glu
20 25 30
Tyr Ala Asn Thr Ile Ala Leu Pro Pro Arg Thr Gln Ser Leu Cys Leu
35 40 45
Val Val Cys Leu Asp Glu Lys Glu Gly Lys Thr Gln Glu Leu Lys Asn
50 55 60
Ile Arg Thr Asn Ser Glu Leu Leu Lys Glu Arg Ile Ile Ala Ala Phe
65 70 75 80
His Glu Gly Lys Asn Leu Lys Thr Tyr His Glu Lys Lys Lys Gly Asp
85 90 95
Asp Gly Lys Lys Leu Cys Lys Asp Leu Lys Tyr Ser Phe Ala Asp Tyr
100 105 110
Gly Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Asp Phe Thr Lys
115 120 125
Asp Leu Glu Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe Arg Lys
130 135 140
Tyr Ile Lys Lys Asn Asn Thr Ala Glu Gln His Thr Ser Tyr Ser Ser
145 150 155 160
Leu Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile
165 170 175
Trp Thr Ala Met Lys His Gly Ala Glu Met Asn Gly Thr Thr Cys Ser
180 185 190
Cys Ser Gly Asp Ser Ser Asn Asp Ile Pro Thr Ile Asp Leu Ile Pro
195 200 205
Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His Phe Cys Glu Gln
210 215 220
Arg Gln Ala Lys Val Asn Ala Val Ile Lys Asn Cys Lys Ser Cys Lys
225 230 235 240
Glu Cys Gly Gly Thr Cys Asn Gly Glu Cys Lys Thr Glu Cys Lys Thr
245 250 255
Lys Cys Lys Gly Glu Cys Glu Lys Tyr Lys Glu Phe Ile Glu Lys Cys
260 265 270
Glu Gly Gln Ala Ala Glu Gly Thr Ser Gly Ser Ser Trp Ser Lys Arg
275 280 285
Trp Tyr Gln Ile Tyr Met Arg Tyr Ser Lys Tyr Ile Glu Asp Ala Lys
290 295 300
Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Thr Ser Ser Gly Ala
305 310 315 320
Asn Ser Gly Val Thr Thr Thr Glu Asn Lys Cys Val Gln Ser
325 330
<210> 26
<211> 351
<212> PRT
<213> 恶性疟原虫
<400> 26
Lys Cys Asp Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Lys Asn
1 5 10 15
Trp Ile Trp Lys Lys Tyr Ser Gly Thr Glu Gly Gly Leu Gln Lys Glu
20 25 30
Tyr Ala Asn Thr Ile Ala Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu
35 40 45
Val Cys Leu His Glu Lys Glu Glu Lys Thr Gln Glu Leu Lys Asn Ile
50 55 60
Ser Thr Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His
65 70 75 80
Glu Gly Lys Asn Leu Lys Ile Ser Pro Gln Asn Lys Asn Asp Asn Gly
85 90 95
Lys Asn Leu Cys Lys Asp Leu Lys Tyr Ser Phe Ala Asp Tyr Gly Asp
100 105 110
Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Asp Phe Thr Lys Asp Leu
115 120 125
Glu Leu Asn Leu Gln Gln Ile Phe Gly Lys Leu Phe Arg Lys Tyr Ile
130 135 140
Lys Lys Asn Asn Thr Ala Glu Gln Asp Thr Leu Tyr Ser Ser Leu Asp
145 150 155 160
Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Thr
165 170 175
Ala Met Lys His Gly Ala Gly Met Asn Gly Thr Thr Cys Cys Gly Asp
180 185 190
Gly Ser Val Thr Gly Ser Ser Asp Ser Gly Ser Thr Thr Cys Cys Gly
195 200 205
Asp Gly Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp Ile Pro Thr
210 215 220
Ile Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu
225 230 235 240
His Phe Cys Glu Gln Arg Gln Ala Lys Val Lys Asp Val Ile Lys Asn
245 250 255
Cys Asn Ser Cys Lys Glu Cys Gly Gly Thr Cys Asn Gly Glu Cys Lys
260 265 270
Thr Glu Cys Glu Lys Lys Cys Lys Gly Glu Cys Glu Ala Tyr Lys Lys
275 280 285
Phe Ile Glu Lys Cys Asn Gly Gly Gly Gly Glu Gly Thr Ser Gly Ser
290 295 300
Ser Trp Ser Lys Arg Trp Asp Gln Ile Tyr Met Arg Tyr Ser Lys Tyr
305 310 315 320
Ile Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly
325 330 335
Thr Ser Ser Thr Thr Asn Ala Ala Glu Asn Lys Cys Val Gln Ser
340 345 350
<210> 27
<211> 353
<212> PRT
<213> 恶性疟原虫
<400> 27
Lys Cys Asp Lys Cys Lys Ser Gly Thr Ser Thr Val Asn Lys Lys Trp
1 5 10 15
Ile Trp Lys Lys Phe Pro Gly Lys Glu Gly Gly Leu Gln Glu Glu Tyr
20 25 30
Ala Asn Thr Ile Ala Leu Pro Pro Arg Thr Gln Ser Leu Cys Leu Val
35 40 45
Val Cys Leu Asp Glu Lys Glu Gly Lys Thr Gln His Lys Thr Ile Ser
50 55 60
Thr Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu
65 70 75 80
Gly Lys Asn Leu Lys Ile Ser Asn Lys Lys Lys Asn Asp Glu Asn Asn
85 90 95
Ser Lys Leu Cys Lys Asp Leu Lys Tyr Ser Phe Ala Asp Tyr Gly Asp
100 105 110
Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Asp Phe Thr Lys Asp Leu
115 120 125
Glu Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe Arg Lys Tyr Ile
130 135 140
Lys Lys Asn Asn Thr Ala Glu Gln Asp Thr Ser Tyr Ser Ser Leu Asp
145 150 155 160
Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Leu
165 170 175
Ala Met Lys His Gly Thr Thr Cys Ser Ser Gly Ser Gly Asp Asn Gly
180 185 190
Asp Gly Ser Val Thr Gly Ser Ser Asp Ser Gly Ser Thr Thr Cys Cys
195 200 205
Gly Asp Gly Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp Ile Pro
210 215 220
Thr Ile Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val
225 230 235 240
Glu His Phe Cys Lys Gln Arg Gln Ala Lys Val Lys Asp Val Ile Glu
245 250 255
Asn Cys Lys Ser Cys Lys Asn Thr Ser Ser Lys Thr Lys Leu Gly Asp
260 265 270
Thr Cys Asn Ser Asp Cys Lys Thr Lys Cys Lys Val Ala Cys Glu Lys
275 280 285
Tyr Lys Glu Phe Ile Glu Lys Cys Val Ser Ala Ala Gly Gly Thr Ser
290 295 300
Gly Ser Ser Trp Val Lys Arg Trp Asp Gln Ile Tyr Met Arg Tyr Ser
305 310 315 320
Lys Tyr Ile Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn
325 330 335
Cys Gly Pro Ser Ser Thr Thr Ser Thr Ala Glu Ser Lys Cys Val Gln
340 345 350
Ser
<210> 28
<211> 327
<212> PRT
<213> 恶性疟原虫
<400> 28
Lys Cys Asp Lys Cys Lys Ser Gly Thr Ser Thr Val Asn Lys Lys Trp
1 5 10 15
Ile Trp Lys Lys Tyr Ser Gly Lys Glu Gly Gly Leu Gln Lys Glu Tyr
20 25 30
Ala Asn Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Cys Leu Val
35 40 45
Cys Leu His Glu Lys Glu Gly Lys Thr Gln Glu Leu Lys Asn Ile Ser
50 55 60
Thr Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu
65 70 75 80
Gly Lys Asn Leu Lys Ile Ser Asn Lys Lys Lys Asn Asp Asp Asn Gly
85 90 95
Lys Lys Leu Cys Lys Asp Leu Lys Tyr Ser Phe Ala Asp Tyr Gly Asp
100 105 110
Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Asp Phe Thr Lys Asp Leu
115 120 125
Glu Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe Arg Lys Tyr Ile
130 135 140
Lys Lys Asn Asn Thr Ala Glu Gln Asp Thr Leu Tyr Ser Ser Leu Asp
145 150 155 160
Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Thr
165 170 175
Ala Met Lys His Gly Ala Gly Met Asn Ser Thr Thr Cys Ser Cys Ser
180 185 190
Gly Asp Ser Ser Asn Asp Ile Pro Thr Ile Asp Leu Ile Pro Gln Tyr
195 200 205
Leu Arg Phe Leu Gln Glu Trp Val Glu His Phe Cys Lys Gln Arg Gln
210 215 220
Glu Lys Val Asn Ala Val Ile Thr Asn Cys Lys Ser Cys Lys Glu Ser
225 230 235 240
Gly Gly Thr Cys Asn Ser Asp Cys Glu Lys Lys Cys Lys Ile Glu Cys
245 250 255
Glu Lys Tyr Lys Asn Phe Ile Glu Lys Cys Val Thr Ala Ala Gly Gly
260 265 270
Thr Ser Gly Ser Ser Trp Ser Lys Arg Trp Asp Gln Ile Tyr Lys Met
275 280 285
Tyr Ser Lys Tyr Ile Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr
290 295 300
Lys Asn Cys Gly Pro Ser Ser Thr Thr Asn Ala Ala Ala Ser Thr Asp
305 310 315 320
Glu Asn Lys Cys Val Gln Ser
325
<210> 29
<211> 628
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 29
Asn Tyr Ile Lys Gly Asp Pro Tyr Phe Ala Glu Tyr Ala Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Ser Ser Asp Thr Glu Asn Ala Ser Glu Thr Pro
20 25 30
Ser Lys Tyr Tyr Asp Glu Ala Cys Asn Cys Asn Glu Ser Glu Ile Ala
35 40 45
Ser Val Gly Gln Ala Gln Thr Ser Gly Pro Ser Ser Asn Lys Thr Cys
50 55 60
Ile Thr His Ser Ser Ile Lys Thr Asn Lys Lys Lys Glu Cys Lys Asp
65 70 75 80
Val Lys Leu Gly Ile Asn Asn Asn Asp Lys Val Leu Arg Val Cys Val
85 90 95
Ile Glu Asp Thr Ser Leu Ser Gly Val Asp Asn Cys Cys Cys Gln Asp
100 105 110
Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Arg Gly Ser
115 120 125
Ser Ser Asn Gly Ser Cys Asp Lys Asn Ser Glu Glu Ile Cys Gln Lys
130 135 140
Lys Leu Glu Lys Val Phe Ala Ser Leu Thr Asn Gly Tyr Lys Cys Asp
145 150 155 160
Lys Cys Lys Ser Gly Thr Ser Arg Ser Lys Lys Lys Trp Ile Trp Lys
165 170 175
Lys Ser Ser Gly Asn Glu Glu Gly Leu Gln Lys Glu Tyr Ala Asn Thr
180 185 190
Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Cys Leu Val Cys Leu His
195 200 205
Glu Lys Glu Gly Lys Thr Gln His Lys Thr Ile Ser Thr Asn Ser Glu
210 215 220
Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu Gly Lys Asn Leu
225 230 235 240
Lys Thr Ser His Glu Lys Lys Asn Asp Asp Asn Gly Lys Lys Leu Cys
245 250 255
Lys Ala Leu Glu Tyr Ser Phe Ala Asp Tyr Gly Asp Leu Ile Lys Gly
260 265 270
Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp Leu Glu Leu Asn Leu
275 280 285
Gln Lys Ile Phe Gly Lys Leu Phe Arg Lys Tyr Ile Lys Lys Asn Asn
290 295 300
Thr Ala Glu Gln His Thr Ser Tyr Ser Ser Leu Asp Glu Leu Arg Glu
305 310 315 320
Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Thr Ala Met Lys His
325 330 335
Gly Ala Gly Met Asn Gly Thr Thr Cys Ser Cys Ser Gly Asp Ser Ser
340 345 350
Asn Asp Met Pro Thr Ile Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu
355 360 365
Gln Glu Trp Val Glu His Phe Cys Lys Gln Arg Gln Glu Lys Val Asn
370 375 380
Ala Val Ile Glu Asn Cys Asn Ser Cys Lys Glu Ser Gly Gly Thr Cys
385 390 395 400
Asn Ser Asp Cys Lys Thr Glu Cys Lys Asn Lys Cys Glu Ala Tyr Lys
405 410 415
Glu Phe Ile Glu Asp Cys Lys Gly Gly Gly Thr Gly Thr Ala Gly Ser
420 425 430
Pro Trp Ser Lys Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys His
435 440 445
Ile Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly
450 455 460
Thr Ser Ser Thr Thr Asn Ala Ala Ala Ser Thr Asp Glu Asn Lys Cys
465 470 475 480
Val Gln Ser Asp Val Asp Ser Phe Phe Lys His Leu Ile Asp Ile Gly
485 490 495
Leu Thr Thr Pro Ser Ser Tyr Leu Ser Asn Val Leu Asp Asp Asn Ile
500 505 510
Cys Gly Ala Asp Lys Ala Pro Trp Thr Thr Tyr Thr Thr Tyr Thr Thr
515 520 525
Thr Lys Asn Cys Asp Ile Gln Lys Lys Thr Pro Lys Ser Gln Ser Cys
530 535 540
Asp Thr Leu Val Val Val Asn Val Pro Ser Pro Leu Gly Asn Thr Pro
545 550 555 560
His Glu Tyr Lys Tyr Ala Cys Glu Cys Lys Ile Pro Thr Thr Glu Glu
565 570 575
Thr Cys Asp Asp Arg Lys Glu Tyr Met Asn Gln Trp Ser Cys Gly Ser
580 585 590
Ala Gln Thr Val Arg Gly Arg Ser Gly Lys Asp Asp Tyr Glu Leu Tyr
595 600 605
Thr Tyr Asn Gly Val Lys Glu Thr Lys Pro Leu Gly Thr Leu Lys Asn
610 615 620
Ser Lys Leu Asp
625
<210> 30
<211> 350
<212> PRT
<213> 恶性疟原虫
<400> 30
Lys Cys Glu Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Lys Asn
1 5 10 15
Trp Ile Trp Arg Lys Phe Arg Gly Thr Glu Gly Gly Leu Gln Glu Glu
20 25 30
Tyr Ala Asn Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Cys Leu
35 40 45
Val Val Cys Leu Asp Glu Lys Gly Lys Lys Thr Gln Glu Leu Lys Asn
50 55 60
Ile Arg Thr Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe
65 70 75 80
His Glu Gly Lys Asn Leu Lys Pro Ser His Gln Asn Lys Asn Ser Gly
85 90 95
Asn Lys Glu Asn Leu Cys Lys Ala Leu Lys Tyr Ser Phe Ala Asp Tyr
100 105 110
Gly Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Asp Phe Thr Lys
115 120 125
Asp Leu Glu Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe Arg Lys
130 135 140
Tyr Ile Lys Lys Asn Asn Thr Ala Glu Gln His Thr Ser Tyr Ser Ser
145 150 155 160
Leu Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile
165 170 175
Trp Thr Ala Met Lys His Gly Ala Glu Met Asn Gly Thr Thr Cys Asn
180 185 190
Ala Asp Gly Ser Val Thr Gly Ser Ser Asp Ser Gly Ser Thr Thr Cys
195 200 205
Ser Gly Asp Asn Gly Ser Ile Ser Cys Asp Asp Ile Pro Thr Ile Asp
210 215 220
Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His Phe
225 230 235 240
Cys Lys Gln Arg Gln Glu Lys Val Asn Ala Val Ile Asn Ser Cys Asn
245 250 255
Ser Cys Lys Asn Thr Ser Ser Lys Thr Lys Leu Gly Asp Thr Cys Asn
260 265 270
Ser Asp Cys Lys Thr Lys Cys Lys Ile Glu Cys Glu Lys Tyr Lys Thr
275 280 285
Phe Ile Glu Lys Cys Val Thr Ala Ala Gly Gly Thr Ser Gly Ser Pro
290 295 300
Trp Ser Lys Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys Tyr Ile
305 310 315 320
Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Pro
325 330 335
Ser Ser Thr Thr Ser Thr Ala Glu Ser Lys Cys Val Gln Ser
340 345 350
<210> 31
<211> 330
<212> PRT
<213> 恶性疟原虫
<400> 31
Lys Cys Asp Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Lys Asn
1 5 10 15
Trp Ile Trp Arg Lys Tyr Ser Gly Asn Gly Glu Gly Leu Gln Lys Glu
20 25 30
Tyr Ala Asn Thr Ile Gly Leu Pro Pro Arg Thr His Ser Leu Tyr Leu
35 40 45
Val Cys Leu His Glu Lys Glu Gly Lys Thr Gln Glu Leu Lys Asn Ile
50 55 60
Arg Thr Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His
65 70 75 80
Glu Gly Lys Asn Leu Lys Thr Thr Tyr Leu Glu Asn Lys Asn Asp Glu
85 90 95
Asn Lys Lys Lys Leu Cys Lys Ala Leu Lys Tyr Ser Phe Ala Asp Tyr
100 105 110
Gly Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Asp Phe Thr Lys
115 120 125
Asp Leu Glu Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe Arg Lys
130 135 140
Tyr Ile Lys Lys Asn Ile Ala Ser Asp Glu Asn Thr Leu Tyr Ser Ser
145 150 155 160
Leu Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile
165 170 175
Trp Thr Ala Met Lys His Gly Ala Glu Met Asn Gly Thr Thr Cys Ser
180 185 190
Ser Gly Ser Gly Asp Asn Gly Ser Ile Ser Cys Asp Asp Ile Pro Thr
195 200 205
Ile Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Gly
210 215 220
His Phe Cys Lys Gln Arg Gln Glu Lys Val Asn Ala Val Ile Thr Asn
225 230 235 240
Cys Asn Ser Cys Lys Glu Ser Gly Gly Thr Cys Asn Ser Asp Cys Glu
245 250 255
Lys Lys Cys Lys Ile Glu Cys Glu Lys Tyr Lys Lys Phe Ile Glu Glu
260 265 270
Cys Arg Thr Ala Ala Gly Gly Thr Ser Gly Ser Pro Trp Ser Lys Arg
275 280 285
Trp Asp Gln Ile Tyr Lys Met Tyr Ser Lys Tyr Ile Glu Asp Ala Lys
290 295 300
Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Pro Ser Ser Thr Thr
305 310 315 320
Ser Thr Ala Glu Ser Lys Cys Val Gln Ser
325 330
<210> 32
<211> 334
<212> PRT
<213> 恶性疟原虫
<400> 32
Lys Cys Asp Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Lys Asn
1 5 10 15
Trp Ile Trp Arg Lys Tyr Ser Gly Asn Gly Glu Gly Leu Gln Lys Glu
20 25 30
Tyr Ala Asn Thr Ile Gly Leu Pro Pro Arg Thr His Ser Leu Tyr Leu
35 40 45
Val Cys Leu His Glu Lys Glu Gly Lys Thr Gln His Lys Thr Ile Ser
50 55 60
Thr Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu
65 70 75 80
Gly Lys Asn Leu Lys Lys Arg Tyr Pro Gln Asn Asn Asn Ser Gly Asn
85 90 95
Lys Lys Lys Leu Cys Lys Asp Leu Lys Tyr Ser Phe Ala Asp Tyr Gly
100 105 110
Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp
115 120 125
Leu Glu Leu Asn Leu Gln Lys Ala Phe Gly Lys Leu Phe Arg Lys Tyr
130 135 140
Ile Lys Lys Asn Ile Ala Ser Asp Glu Asn Thr Leu Tyr Ser Ser Leu
145 150 155 160
Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp
165 170 175
Leu Ala Met Lys His Gly Ala Glu Met Asn Gly Thr Met Cys Asn Ala
180 185 190
Asp Gly Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp Met Ser Thr
195 200 205
Ile Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu
210 215 220
His Phe Cys Glu Gln Arg Gln Ala Lys Val Lys Asp Val Ile Asn Ser
225 230 235 240
Cys Lys Ser Cys Lys Glu Ser Gly Asp Thr Cys Asn Ser Asp Cys Glu
245 250 255
Lys Lys Cys Lys Asn Lys Cys Asp Ala Tyr Lys Thr Phe Ile Glu Glu
260 265 270
Phe Cys Thr Ala Asp Gly Gly Thr Ala Gly Ser Pro Trp Ser Lys Arg
275 280 285
Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys Tyr Ile Glu Asp Ala Lys
290 295 300
Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Thr Ser Ser Gly Ala
305 310 315 320
Asn Ser Gly Val Thr Thr Thr Glu Asn Lys Cys Val Gln Ser
325 330
<210> 33
<211> 350
<212> PRT
<213> 恶性疟原虫
<400> 33
Lys Cys Asp Lys Cys Lys Ser Gly Thr Ser Thr Val Asn Lys Asn Trp
1 5 10 15
Ile Trp Lys Lys Tyr Ser Gly Lys Glu Glu Gly Leu Gln Lys Glu Tyr
20 25 30
Ala Asn Thr Ile Ala Leu Pro Pro Arg Thr His Ser Leu Tyr Leu Val
35 40 45
Cys Leu His Glu Lys Gly Lys Lys Thr Gln Glu Leu Lys Asn Ile Arg
50 55 60
Thr Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu
65 70 75 80
Gly Lys Asn Leu Lys Thr Ser Pro Gln Asn Asn Asn Ser Gly Asn Lys
85 90 95
Lys Lys Leu Cys Lys Ala Leu Lys Tyr Ser Phe Ala Asp Tyr Gly Asp
100 105 110
Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Asp Phe Thr Lys Asp Leu
115 120 125
Glu Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe Arg Lys Tyr Ile
130 135 140
Lys Lys Asn Asn Thr Ala Glu Gln His Thr Ser Tyr Ser Ser Leu Asp
145 150 155 160
Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Leu
165 170 175
Ala Met Lys His Gly Ala Glu Met Asn Gly Thr Thr Cys Cys Gly Asp
180 185 190
Gly Ser Val Thr Gly Ser Ser Asp Ser Gly Ser Thr Thr Cys Ser Gly
195 200 205
Asp Asn Gly Ser Ile Ser Cys Asp Asp Met Pro Thr Thr Asp Phe Ile
210 215 220
Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His Phe Cys Lys
225 230 235 240
Gln Arg Gln Glu Lys Val Lys His Val Met Glu Ser Cys Lys Ser Cys
245 250 255
Lys Glu Cys Gly Asp Thr Cys Asn Gly Glu Cys Lys Thr Glu Cys Glu
260 265 270
Lys Lys Cys Lys Asn Lys Cys Glu Ala Tyr Lys Thr Phe Ile Glu Lys
275 280 285
Cys Val Ser Ala Asp Gly Gly Thr Ser Gly Ser Ser Trp Ser Lys Arg
290 295 300
Trp Asp Gln Ile Tyr Met Arg Tyr Ser Lys Tyr Ile Glu Asp Ala Lys
305 310 315 320
Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Thr Ser Ser Thr Thr
325 330 335
Asn Ala Ala Ala Ser Thr Ala Glu Asn Lys Cys Val Gln Ser
340 345 350
<210> 34
<211> 647
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 34
Asp Tyr Ile Lys Asp Asp Pro Tyr Ser Ala Glu Tyr Ala Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Pro Ser Asp Ala Asn Thr Ser Ser Gly Glu Thr
20 25 30
Ala Asn His Asn Asp Glu Val Cys Asn Cys Asn Glu Ser Glu Ile Ala
35 40 45
Ser Val Glu Leu Ala Pro Ile Ser Asp Ser Ser Ser Asn Lys Thr Cys
50 55 60
Ile Thr His Ser Phe Ile Gly Ala Asn Lys Lys Lys Glu Cys Lys Asp
65 70 75 80
Val Lys Leu Gly Val Arg Glu Lys Asp Lys Asp Leu Lys Ile Cys Val
85 90 95
Ile Glu Asp Asp Ser Leu Arg Gly Val Glu Asn Cys Cys Cys Gln Asp
100 105 110
Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Ser Gly Ser
115 120 125
Ser Ser Asn Gly Ser Cys Asp Lys Asn Ser Glu Asp Glu Cys Gln Lys
130 135 140
Lys Leu Glu Asn Val Phe Ala Ser Leu Lys Asn Gly Tyr Lys Cys Asp
145 150 155 160
Lys Cys Lys Ser Gly Thr Ser Thr Val Asn Lys Lys Trp Ile Trp Arg
165 170 175
Lys Tyr Ser Gly Asn Gly Glu Gly Leu Gln Lys Glu Tyr Ala Asn Thr
180 185 190
Ile Gly Leu Pro Pro Arg Thr His Ser Leu Tyr Leu Val Cys Leu His
195 200 205
Glu Lys Glu Gly Lys Thr Gln His Lys Thr Ile Ser Thr Asn Ser Glu
210 215 220
Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu Gly Lys Asn Leu
225 230 235 240
Lys Thr Ser His Gln Asn Asn Asn Ser Gly Asn Lys Lys Lys Leu Cys
245 250 255
Lys Ala Leu Lys Tyr Ser Phe Ala Asp Tyr Gly Asp Leu Ile Lys Gly
260 265 270
Thr Ser Ile Trp Asp Asn Asp Phe Thr Lys Asp Leu Glu Leu Asn Leu
275 280 285
Gln Lys Ile Phe Gly Lys Leu Phe Arg Lys Tyr Ile Lys Lys Asn Ile
290 295 300
Ala Ser Asp Glu Asn Thr Ser Tyr Ser Ser Leu Asp Glu Leu Arg Glu
305 310 315 320
Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Leu Ala Met Lys His
325 330 335
Gly Ala Glu Met Asn Ser Thr Met Cys Asn Gly Asp Gly Ser Val Thr
340 345 350
Gly Ser Ser Asp Ser Gly Ser Thr Thr Cys Ser Gly Asp Asn Gly Ser
355 360 365
Ile Ser Cys Asp Asp Ile Pro Thr Ile Asp Leu Ile Pro Gln Tyr Leu
370 375 380
Arg Phe Leu Gln Glu Trp Val Glu His Phe Cys Lys Gln Arg Gln Glu
385 390 395 400
Lys Val Lys Asp Val Ile Thr Asn Cys Lys Ser Cys Lys Glu Ser Gly
405 410 415
Asp Thr Cys Asn Ser Asp Cys Glu Lys Lys Cys Lys Asn Lys Cys Glu
420 425 430
Ala Tyr Lys Lys Phe Ile Glu Glu Arg Arg Thr Ala Ala Gln Gly Thr
435 440 445
Ala Glu Ser Ser Trp Val Lys Arg Trp Asp Gln Ile Tyr Met Arg Tyr
450 455 460
Ser Lys Tyr Ile Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys
465 470 475 480
Ser Cys Gly Pro Ser Ser Thr Thr Asn Ala Ala Ala Ser Thr Ala Glu
485 490 495
Asn Lys Cys Val Gln Ser Asp Ile Asp Ser Phe Phe Lys His Leu Ile
500 505 510
Asp Ile Gly Leu Thr Thr Pro Ser Ser Tyr Leu Ser Ile Val Leu Asp
515 520 525
Asp Asn Ile Cys Gly Ala Asp Asn Ala Pro Trp Thr Thr Tyr Thr Thr
530 535 540
Tyr Thr Thr Thr Lys Asn Cys Asp Ile Lys Lys Lys Thr Pro Lys Pro
545 550 555 560
Gln Ser Cys Asp Thr Leu Val Val Val Asn Val Pro Ser Pro Leu Gly
565 570 575
Asn Thr Pro His Glu Tyr Lys Tyr Ala Cys Gln Cys Arg Thr Pro Asn
580 585 590
Lys Gln Glu Ser Cys Asp Asp Arg Lys Glu Tyr Met Asn Gln Trp Ser
595 600 605
Ser Gly Ser Ala Gln Thr Val Arg Gly Arg Ser Thr Asn Asn Asp Tyr
610 615 620
Glu Leu Tyr Thr Tyr Asn Gly Val Lys Glu Thr Lys Pro Leu Gly Thr
625 630 635 640
Leu Lys Asn Ser Lys Leu Asp
645
<210> 35
<211> 341
<212> PRT
<213> 恶性疟原虫
<400> 35
Lys Cys Asp Lys Cys Lys Ser Gly Thr Ser Thr Val Asn Lys Lys Trp
1 5 10 15
Ile Trp Arg Lys Ser Ser Gly Asn Lys Glu Gly Leu Gln Lys Glu Tyr
20 25 30
Ala Asn Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu Gly
35 40 45
Asn Leu Pro Lys Leu Glu Asn Val Cys Glu Asp Val Lys Asp Ile Asn
50 55 60
Phe Asp Thr Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Val Ser Phe
65 70 75 80
His Glu Gly Lys Asn Leu Lys Thr Ser His Glu Lys Lys Asn Asp Asp
85 90 95
Asn Gly Lys Lys Leu Cys Lys Ala Leu Glu Tyr Ser Phe Ala Asp Tyr
100 105 110
Gly Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys
115 120 125
Asp Leu Glu Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe Arg Lys
130 135 140
Tyr Ile Lys Lys Asn Asn Thr Ala Glu Gln Asp Thr Ser Tyr Ser Ser
145 150 155 160
Leu Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile
165 170 175
Trp Thr Ala Met Lys His Gly Ala Gly Met Asn Ile Thr Thr Cys Cys
180 185 190
Gly Asp Gly Ser Ser Gly Glu Asn Gln Thr Asn Ser Cys Asp Asp Ile
195 200 205
Pro Thr Ile Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp
210 215 220
Val Glu His Phe Cys Lys Gln Arg Gln Glu Lys Val Asn Ala Val Val
225 230 235 240
Thr Asn Cys Lys Ser Cys Lys Glu Ser Gly Gly Thr Cys Asn Gly Glu
245 250 255
Cys Lys Thr Lys Cys Lys Asn Lys Cys Glu Val Tyr Lys Thr Phe Ile
260 265 270
Asp Asn Val Gly Asp Gly Thr Ala Gly Ser Pro Trp Val Lys Arg Trp
275 280 285
Asp Gln Ile Tyr Lys Arg Tyr Ser Lys His Ile Glu Asp Ala Lys Arg
290 295 300
Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Ile Thr Thr Gly Thr Ile
305 310 315 320
Ser Gly Glu Ser Ser Gly Ala Thr Ser Gly Val Thr Thr Thr Glu Asn
325 330 335
Lys Cys Val Gln Ser
340
<210> 36
<211> 632
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 36
Asn Tyr Ile Lys Asp Asp Pro Tyr Ser Lys Glu Tyr Val Thr Lys Leu
1 5 10 15
Ser Phe Ile Pro Asn Ser Ser Asp Ala Asn Thr Ser Ser Glu Lys Ile
20 25 30
Gln Lys Asn Asn Asp Glu Val Cys Asn Pro Asn Glu Ser Gly Ile Ser
35 40 45
Ser Val Glu Gln Ala Gln Thr Ser Gly Pro Ser Ser Asn Lys Thr Cys
50 55 60
Ile Thr His Ser Ser Ile Lys Ala Asn Lys Lys Lys Glu Cys Lys Asp
65 70 75 80
Val Lys Leu Gly Val Arg Glu Asn Asp Lys Asp Leu Lys Ile Cys Val
85 90 95
Ile Glu Asp Thr Ser Leu Ser Gly Val Asp Asn Cys Cys Cys Gln Asp
100 105 110
Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Arg Gly Ser
115 120 125
Ser Ser Asn Asp Ser Cys Asp Asn Lys Asn Gln Asp Glu Cys Gln Lys
130 135 140
Lys Leu Asp Glu Ala Leu Glu Ser Leu His Asn Gly Tyr Lys Asn Gln
145 150 155 160
Lys Cys Lys Ser Gly Thr Ser Thr Val Asn Lys Lys Trp Ile Trp Lys
165 170 175
Lys Ser Ser Gly Asn Lys Glu Gly Leu Gln Lys Glu Tyr Ala Asn Thr
180 185 190
Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu Gly Asn Leu Pro
195 200 205
Lys Leu Glu Asn Val Ser Lys Gly Val Thr Asp Ile Ile Tyr Asp Thr
210 215 220
Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Val Ser Phe His Glu Gly
225 230 235 240
Lys Asn Leu Lys Thr Ser His Glu Lys Lys Asn Asp Asp Asn Gly Lys
245 250 255
Lys Leu Cys Lys Ala Leu Glu Tyr Ser Phe Ala Asp Tyr Gly Asp Leu
260 265 270
Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp Leu Glu
275 280 285
Leu Asn Leu Gln Lys Ala Phe Gly Lys Leu Phe Arg Lys Tyr Ile Lys
290 295 300
Lys Asn Ile Ser Ala Glu Gln Asp Thr Ser Tyr Ser Ser Leu Asp Glu
305 310 315 320
Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Ile Ala
325 330 335
Met Lys His Gly Ala Gly Met Asn Gly Thr Thr Cys Cys Gly Asp Gly
340 345 350
Ser Ser Gly Glu Asn Gln Thr Asn Ser Cys Asp Asp Ile Pro Thr Ile
355 360 365
Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His
370 375 380
Phe Cys Glu Gln Arg Gln Ala Lys Val Lys Asp Val Ile Thr Asn Cys
385 390 395 400
Lys Ser Cys Lys Asn Thr Ser Gly Glu Arg Lys Ile Gly Gly Thr Cys
405 410 415
Asn Gly Glu Cys Lys Thr Lys Cys Lys Asn Lys Cys Glu Ala Tyr Lys
420 425 430
Thr Phe Ile Glu His Cys Lys Gly Gly Asp Gly Thr Ala Gly Ser Ser
435 440 445
Trp Val Lys Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys His Ile
450 455 460
Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys Ser Cys Gly Thr
465 470 475 480
Ser Thr Ala Glu Asn Lys Cys Val Gln Ser Asp Ile Asp Ser Phe Phe
485 490 495
Lys His Leu Ile Asp Ile Gly Leu Thr Thr Pro Ser Ser Tyr Leu Ser
500 505 510
Ile Val Leu Asp Glu Asn Asn Cys Gly Glu Asp Lys Ala Pro Trp Thr
515 520 525
Thr Tyr Thr Thr Thr Lys Asn Cys Asp Ile Gln Lys Asp Lys Ser Lys
530 535 540
Ser Gln Ser Ser Asp Thr Leu Val Val Val Asn Val Pro Ser Pro Leu
545 550 555 560
Gly Asn Thr Pro His Gly Tyr Lys Tyr Ala Cys Gln Cys Lys Ile Pro
565 570 575
Thr Thr Glu Glu Thr Cys Asp Asp Arg Lys Glu Tyr Met Asn Gln Trp
580 585 590
Ser Cys Gly Ser Ala Arg Thr Met Lys Arg Gly Tyr Lys Asn Asp Asn
595 600 605
Tyr Glu Leu Cys Lys Tyr Asn Gly Val Asp Val Lys Pro Thr Thr Val
610 615 620
Arg Ser Ser Ser Thr Lys Leu Asp
625 630
<210> 37
<211> 639
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 37
Asp Tyr Ile Lys Gly Asp Pro Tyr Ser Ala Glu Tyr Val Thr Lys Leu
1 5 10 15
Ser Phe Ile Pro Asn Ser Ser Asp Ala Asn Asn Pro Ser Glu Lys Ile
20 25 30
Gln Lys Asn Asn Asp Glu Val Cys Asn Cys Asn Glu Ser Glu Ile Ser
35 40 45
Ser Val Gly Gln Ala Ser Ile Ser Asp Pro Ser Ser Asn Lys Thr Cys
50 55 60
Asn Thr His Ser Ser Ile Lys Ala Asn Lys Lys Lys Val Cys Lys Asp
65 70 75 80
Val Lys Leu Gly Val Arg Glu Asn Asp Lys Val Leu Lys Ile Cys Val
85 90 95
Ile Glu His Thr Ser Leu Arg Gly Val Asp Asn Cys Cys Phe Lys Asp
100 105 110
Leu Leu Gly Ile Leu Gln Glu Pro Arg Ile Asp Lys Asn Gln Ser Gly
115 120 125
Ser Ser Ser Asn Gly Ser Cys Asp Lys Asn Ser Glu Glu Ala Cys Glu
130 135 140
Lys Asn Leu Glu Lys Val Leu Ala Ser Leu Thr Asn Gly Tyr Lys Cys
145 150 155 160
Asp Lys Cys Lys Ser Gly Thr Ser Arg Ser Lys Lys Lys Trp Ile Trp
165 170 175
Lys Lys Tyr Ser Gly Lys Glu Gly Gly Leu Gln Glu Glu Tyr Ala Asn
180 185 190
Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Cys Leu Val Val Cys
195 200 205
Leu Asp Glu Lys Glu Gly Lys Thr Gln Glu Leu Lys Asn Ile Ser Thr
210 215 220
Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe Pro Glu Gly
225 230 235 240
Lys Asn Leu Lys Pro Ser Pro Glu Lys Lys Lys Gly Asp Asn Gly Lys
245 250 255
Lys Leu Cys Lys Asp Leu Lys Tyr Ser Phe Ala Asp Tyr Gly Asp Leu
260 265 270
Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp Leu Glu
275 280 285
Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe Arg Lys Tyr Ile Lys
290 295 300
Lys Asn Ile Ala Ser Asp Glu Asn Thr Leu Tyr Ser Ser Leu Asp Glu
305 310 315 320
Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Leu Ala
325 330 335
Met Lys His Gly Ala Gly Met Asn Ser Thr Met Cys Asn Ala Asp Gly
340 345 350
Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp Met Pro Thr Ile Asp
355 360 365
Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His Phe
370 375 380
Cys Lys Gln Arg Gln Glu Lys Val Lys Pro Val Ile Glu Asn Cys Asn
385 390 395 400
Ser Cys Lys Asn Thr Ser Ser Glu Arg Lys Ile Gly Gly Thr Cys Asn
405 410 415
Ser Asp Cys Lys Thr Glu Cys Lys Asn Lys Cys Glu Val Tyr Lys Lys
420 425 430
Phe Ile Glu Asp Cys Lys Gly Gly Asp Gly Thr Ala Gly Ser Ser Trp
435 440 445
Ser Lys Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys Tyr Ile Glu
450 455 460
Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Pro Ser
465 470 475 480
Ser Thr Thr Asn Ala Ala Glu Asn Lys Cys Val Gln Ser Asp Ile Asp
485 490 495
Ser Phe Phe Lys His Leu Ile Asp Ile Gly Leu Thr Thr Pro Ser Ser
500 505 510
Tyr Leu Ser Thr Val Leu Asp Asp Asn Ile Cys Gly Glu Asp Asn Ala
515 520 525
Pro Trp Thr Thr Tyr Thr Thr Tyr Thr Thr Thr Lys Asn Cys Asp Lys
530 535 540
Asp Lys Lys Lys Ser Lys Ser Gln Ser Cys Asp Thr Leu Val Val Val
545 550 555 560
Asn Val Pro Ser Pro Leu Gly Asn Thr Pro His Glu Tyr Lys Tyr Ala
565 570 575
Cys Glu Cys Arg Thr Pro Asn Lys Gln Glu Ser Cys Asp Asp Arg Lys
580 585 590
Glu Tyr Met Asn Gln Trp Ile Ser Asp Asn Thr Lys Asn Pro Lys Gly
595 600 605
Ser Gly Ser Gly Lys Asp Tyr Tyr Glu Leu Tyr Thr Tyr Asn Gly Val
610 615 620
Asp Val Lys Pro Thr Thr Val Arg Ser Ser Ser Thr Lys Leu Asp
625 630 635
<210> 38
<211> 655
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 38
Asp Tyr Ile Lys Gly Asp Pro Tyr Phe Ala Glu Tyr Ala Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Ser Ser Asp Ala Asn Thr Ser Ser Gly Glu Thr
20 25 30
Ala Asn His Asn Asp Glu Ala Cys Asn Cys Asn Glu Ser Glu Ile Ser
35 40 45
Ser Val Glu His Ala Ser Ile Ser Asp Pro Ser Ser Asn Lys Thr Cys
50 55 60
Asn Thr His Ser Ser Ile Lys Ala Asn Lys Lys Lys Val Cys Lys His
65 70 75 80
Val Lys Leu Gly Val Arg Glu Asn Asp Lys Asp Leu Arg Val Cys Val
85 90 95
Ile Glu His Thr Ser Leu Ser Gly Val Glu Asn Cys Cys Phe Lys Asp
100 105 110
Phe Leu Arg Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Ser Gly Ser
115 120 125
Ser Ser Asn Gly Ser Cys Asp Lys Asn Asn Glu Glu Ala Cys Glu Lys
130 135 140
Asn Leu Glu Lys Val Phe Ala Ser Leu Thr Asn Cys Tyr Lys Cys Glu
145 150 155 160
Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Lys Lys Trp Thr Trp
165 170 175
Arg Lys Ser Ser Gly Asn Lys Gly Gly Leu Gln Glu Glu Tyr Ala Asn
180 185 190
Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Cys Leu Val Val Cys
195 200 205
Leu Asp Glu Lys Glu Gly Lys Lys Thr Gln Glu Leu Lys Asn Ile Arg
210 215 220
Thr Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu
225 230 235 240
Gly Lys Asn Leu Lys Pro Ser His Glu Lys Lys Asn Asp Asp Asn Gly
245 250 255
Lys Lys Asn Asp Asp Asn Asn Ser Lys Leu Cys Lys Asp Leu Lys Tyr
260 265 270
Ser Phe Ala Asp Tyr Gly Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp
275 280 285
Asn Glu Tyr Thr Lys Asp Leu Glu Leu Asn Leu Gln Lys Ile Phe Gly
290 295 300
Lys Leu Phe Arg Lys Tyr Ile Lys Lys Asn Ile Ala Ser Asp Glu Asn
305 310 315 320
Thr Leu Tyr Ser Ser Leu Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr
325 330 335
Asn Lys Lys Tyr Ile Trp Leu Ala Met Lys His Gly Ala Glu Met Asn
340 345 350
Gly Thr Thr Cys Asn Ala Asp Gly Ser Val Thr Gly Ser Gly Ser Ser
355 360 365
Cys Asp Asp Ile Pro Thr Ile Asp Leu Ile Pro Gln Tyr Leu Arg Phe
370 375 380
Leu Gln Glu Trp Val Glu His Phe Cys Lys Gln Arg Gln Ala Lys Val
385 390 395 400
Lys Asp Val Ile Glu Asn Cys Lys Ser Cys Lys Glu Ser Gly Asn Lys
405 410 415
Cys Lys Thr Glu Cys Lys Asn Lys Cys Glu Ala Tyr Lys Lys Phe Ile
420 425 430
Glu Asn Cys Lys Gly Gly Asp Gly Thr Ala Gly Ser Ser Trp Val Lys
435 440 445
Arg Trp Asp Gln Ile Tyr Met Arg Tyr Ser Lys Tyr Ile Glu Asp Ala
450 455 460
Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Pro Ser Ser Ile
465 470 475 480
Thr Asn Val Ser Ala Ser Thr Asp Glu Asn Lys Cys Val Gln Ser Asp
485 490 495
Ile Asp Ser Phe Phe Lys His Leu Ile Asp Ile Gly Leu Thr Thr Pro
500 505 510
Ser Ser Tyr Leu Ser Ile Val Leu Asp Asp Asn Ile Cys Gly Asp Asp
515 520 525
Lys Ala Pro Trp Thr Thr Tyr Thr Thr Tyr Thr Thr Tyr Thr Thr Tyr
530 535 540
Thr Thr Tyr Thr Thr Tyr Thr Thr Tyr Thr Thr Thr Lys Asn Cys Asp
545 550 555 560
Lys Glu Arg Asp Lys Ser Lys Ser Gln Ser Cys Asn Thr Ala Val Val
565 570 575
Val Asn Val Pro Ser Pro Leu Gly Asn Thr Pro His Glu Tyr Lys Tyr
580 585 590
Ala Cys Glu Cys Arg Thr Pro Ser Asn Lys Glu Leu Cys Asp Asp Arg
595 600 605
Lys Glu Tyr Met Asn Gln Trp Ser Ser Gly Ser Ala Gln Thr Val Arg
610 615 620
Asp Arg Ser Gly Lys Asp Tyr Tyr Glu Leu Tyr Thr Tyr Asn Gly Val
625 630 635 640
Lys Glu Thr Lys Leu Pro Lys Lys Leu Asn Ser Ser Lys Leu Asp
645 650 655
<210> 39
<211> 347
<212> PRT
<213> 恶性疟原虫
<400> 39
Lys Cys Asp Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Lys Tyr
1 5 10 15
Trp Ile Trp Lys Lys Ser Ser Val Lys Glu Glu Gly Leu Gln Lys Glu
20 25 30
Tyr Ala Asn Thr Ile Ala Leu Pro Pro Arg Thr His Ser Leu Cys Leu
35 40 45
Val Val Cys Leu Asp Glu Lys Gly Lys Lys Thr Gln Glu Leu Lys Asn
50 55 60
Ile Ser Thr Asn Ser Glu Leu Leu Lys Glu Arg Ile Ile Ala Ala Phe
65 70 75 80
His Glu Gly Lys Asn Leu Lys Thr Thr Tyr Leu Glu Lys Lys Asn Ala
85 90 95
Asp Asn Asn Ser Lys Leu Cys Lys Ala Leu Lys Tyr Ser Phe Ala Asp
100 105 110
Tyr Gly Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr
115 120 125
Lys Asp Leu Glu Leu Asn Leu Gln Gln Ile Phe Gly Lys Leu Phe Arg
130 135 140
Lys Tyr Ile Lys Lys Asn Asn Thr Ala Glu Gln His Thr Leu Tyr Ser
145 150 155 160
Ser Leu Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr
165 170 175
Ile Trp Leu Ala Met Lys His Gly Ala Gly Met Asn Gly Thr Thr Cys
180 185 190
Cys Gly Asp Gly Ser Val Thr Gly Ser Ser Asp Ser Gly Ser Thr Thr
195 200 205
Cys Ser Gly Asp Asn Gly Ser Ile Ser Cys Asp Asp Met Pro Thr Thr
210 215 220
Asp Phe Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His
225 230 235 240
Phe Cys Lys Gln Arg Gln Glu Lys Val Lys Asp Val Ile Glu Asn Cys
245 250 255
Asn Ser Cys Lys Asn Asn Leu Gly Lys Thr Glu Ile Asn Glu Lys Cys
260 265 270
Lys Thr Glu Cys Lys Asn Lys Cys Glu Ala Tyr Lys Asn Phe Ile Glu
275 280 285
Lys Phe Cys Thr Ala Asp Gly Gly Thr Ser Gly Ser Pro Trp Ser Lys
290 295 300
Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys Tyr Ile Glu Asp Ala
305 310 315 320
Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Thr Ser Ser Thr
325 330 335
Thr Ser Thr Ala Glu Asn Lys Cys Val Gln Ser
340 345
<210> 40
<211> 335
<212> PRT
<213> 恶性疟原虫
<400> 40
Lys Cys Glu Lys Cys Lys Ser Gly Thr Ser Thr Val Asn Lys Tyr Trp
1 5 10 15
Ile Trp Arg Lys Ser Ser Gly Asn Lys Glu Gly Leu Gln Lys Glu Tyr
20 25 30
Ala Asn Thr Ile Ala Leu Pro Pro Arg Thr His Ser Leu Cys Leu Val
35 40 45
Val Cys Leu Asp Glu Lys Glu Gly Lys Thr Gln Glu Leu Lys Asn Ile
50 55 60
Ser Thr Asn Ser Glu Leu Leu Lys Glu Arg Ile Ile Ala Ala Phe His
65 70 75 80
Glu Gly Glu Asn Leu Lys Thr Ser His Glu Lys Lys Lys Gly Asp Asp
85 90 95
Gly Lys Lys Asn Ala Asp Asn Asn Ser Lys Leu Cys Lys Ala Leu Lys
100 105 110
Tyr Ser Phe Ala Asp Tyr Gly Asp Leu Ile Lys Gly Thr Ser Ile Trp
115 120 125
Asp Asn Glu Tyr Thr Lys Asp Leu Glu Leu Asn Leu Gln Lys Ile Phe
130 135 140
Gly Lys Leu Phe Arg Lys Tyr Ile Lys Lys Asn Ile Ala Ser Asp Glu
145 150 155 160
Asn Thr Ser Tyr Ser Ser Leu Asp Glu Leu Arg Glu Ser Trp Trp Asn
165 170 175
Thr Asn Lys Lys Tyr Ile Trp Leu Ala Met Lys His Gly Ala Gly Met
180 185 190
Asn Gly Thr Thr Cys Ser Cys Ser Gly Asp Ser Ser Asp Asp Met Pro
195 200 205
Thr Thr Asp Phe Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val
210 215 220
Glu His Phe Cys Lys Gln Arg Gln Glu Asn Val Asn Ala Val Ile Glu
225 230 235 240
Asn Cys Asn Ser Cys Lys Glu Cys Gly Gly Thr Cys Asn Ser Asp Cys
245 250 255
Glu Lys Lys Cys Lys Thr Glu Cys Lys Asn Lys Cys Glu Ala Tyr Lys
260 265 270
Asn Phe Ile Glu Lys Phe Cys Thr Ala Asp Gly Gly Thr Ser Gly Tyr
275 280 285
Ser Trp Ser Lys Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys Tyr
290 295 300
Ile Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys Ser Cys Gly
305 310 315 320
Thr Ser Ser Thr Thr Ser Thr Ala Glu Ser Lys Cys Val Gln Ser
325 330 335
<210> 41
<211> 667
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 41
Ser Tyr Val Lys Asn Asn Pro Tyr Ser Lys Glu Tyr Val Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Pro Ser Asp Ala Asn Asn Pro Ser Glu Thr Pro
20 25 30
Ser Lys Tyr Tyr Asp Glu Val Cys Asn Cys Asn Glu Ser Gly Ile Ala
35 40 45
Cys Val Gly Gln Ala Gln Thr Ser Gly Pro Ser Ser Asn Lys Thr Cys
50 55 60
Ile Thr His Ser Phe Ile Gly Ala Asn Lys Lys Lys Val Cys Lys Asp
65 70 75 80
Val Lys Leu Gly Val Arg Glu Lys Asp Lys Asp Leu Lys Ile Cys Val
85 90 95
Ile Glu Asp Thr Tyr Leu Ser Gly Val Asp Asn Cys Cys Phe Lys Asp
100 105 110
Phe Leu Gly Met Leu Gln Glu Asn Cys Ser Asp Asn Lys Ser Gly Ser
115 120 125
Ser Ser Asn Gly Ser Cys Asn Asn Lys Asn Gln Asp Glu Cys Glu Lys
130 135 140
Asn Leu Asp Glu Ala Leu Ala Ser Leu Thr Asn Gly Tyr Lys Cys Glu
145 150 155 160
Lys Cys Lys Ser Gly Thr Ser Thr Val Asn Lys Tyr Trp Ile Trp Arg
165 170 175
Lys Ser Ser Gly Asn Lys Glu Gly Leu Gln Lys Glu Tyr Ala Asn Thr
180 185 190
Ile Ala Leu Pro Pro Arg Thr His Ser Leu Cys Leu Val Val Cys Leu
195 200 205
Asp Glu Lys Glu Gly Lys Thr Gln His Lys Thr Ile Ser Thr Asn Ser
210 215 220
Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu Gly Lys Asn
225 230 235 240
Leu Lys Thr Ser His Glu Lys Lys Lys Gly Asp Asp Gly Lys Lys Asn
245 250 255
Ala Asp Asn Asn Ser Lys Leu Cys Lys Ala Leu Lys Tyr Ser Phe Ala
260 265 270
Asp Tyr Gly Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Asp Phe
275 280 285
Thr Lys Asp Leu Glu Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe
290 295 300
Arg Lys Tyr Ile Lys Lys Asn Ile Ala Ser Asp Glu Asn Thr Ser Tyr
305 310 315 320
Ser Ser Leu Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys
325 330 335
Tyr Ile Trp Leu Ala Met Lys His Gly Ala Gly Met Asn Ser Thr Thr
340 345 350
Cys Cys Gly Asp Gly Ser Val Thr Gly Ser Ser Asp Ser Gly Ser Thr
355 360 365
Thr Cys Cys Gly Asp Gly Ser Val Thr Gly Ser Gly Ser Ser Cys Asp
370 375 380
Asp Met Pro Thr Thr Asp Phe Ile Pro Gln Tyr Leu Arg Phe Leu Gln
385 390 395 400
Glu Trp Val Glu His Phe Cys Lys Gln Arg Gln Glu Asn Val Asn Ala
405 410 415
Val Ile Glu Asn Cys Asn Ser Cys Lys Glu Cys Gly Gly Thr Cys Asn
420 425 430
Ser Asp Cys Glu Lys Lys Cys Lys Thr Glu Cys Lys Gly Glu Cys Asp
435 440 445
Ala Tyr Lys Glu Phe Ile Glu Lys Cys Asn Gly Gly Ala Ala Glu Gly
450 455 460
Thr Ser Gly Ser Ser Trp Ser Lys Arg Trp Asp Gln Ile Tyr Lys Arg
465 470 475 480
Tyr Ser Lys Tyr Ile Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr
485 490 495
Lys Asn Cys Gly Thr Ser Ser Thr Thr Ser Thr Ala Glu Ser Lys Cys
500 505 510
Val Gln Ser Asp Ile Asp Ser Phe Phe Lys His Leu Ile Asp Ile Gly
515 520 525
Leu Thr Thr Pro Ser Ser Tyr Leu Ser Ile Val Leu Asp Glu Asn Ile
530 535 540
Cys Gly Ala Asp Asn Ala Pro Trp Thr Thr Tyr Thr Thr Tyr Thr Thr
545 550 555 560
Tyr Thr Thr Tyr Thr Thr Thr Glu Lys Cys Asn Lys Glu Thr Asp Lys
565 570 575
Ser Lys Leu Gln Gln Cys Asn Thr Ser Val Val Val Asn Val Pro Ser
580 585 590
Pro Leu Gly Asn Thr Pro His Gly Tyr Lys Tyr Val Cys Glu Cys Arg
595 600 605
Thr Pro Asn Lys Gln Glu Thr Cys Asp Asp Arg Lys Glu Tyr Met Asn
610 615 620
Gln Trp Ile Ser Asp Asn Thr Lys Asn Pro Lys Gly Ser Arg Ser Thr
625 630 635 640
Asn Asn Asp Tyr Glu Leu Tyr Thr Tyr Asn Gly Val Gln Ile Lys Pro
645 650 655
Thr Thr Val Arg Ser Asn Ser Thr Lys Leu Asp
660 665
<210> 42
<211> 348
<212> PRT
<213> 恶性疟原虫
<400> 42
Lys Cys Asp Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Lys Asn
1 5 10 15
Trp Ile Trp Lys Lys Ser Ser Gly Asn Glu Lys Gly Leu Gln Lys Glu
20 25 30
Tyr Ala Asn Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Cys Leu
35 40 45
Val Val Cys Leu Asp Glu Lys Glu Gly Lys Thr Gln Glu Leu Lys Asn
50 55 60
Ile Arg Thr Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe
65 70 75 80
His Glu Gly Lys Asn Leu Lys Thr Ser His Glu Lys Lys Lys Gly Asp
85 90 95
Asn Asn Ser Lys Leu Cys Lys Asp Leu Lys Tyr Ser Phe Ala Asp Tyr
100 105 110
Gly Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys
115 120 125
Asp Leu Glu Leu Asn Leu Gln Asn Asn Phe Gly Lys Leu Phe Arg Lys
130 135 140
Tyr Ile Lys Lys Asn Ile Ala Ser Asp Glu Asn Thr Ser Tyr Ser Ser
145 150 155 160
Leu Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile
165 170 175
Trp Leu Ala Met Lys His Gly Ala Gly Met Asn Ser Thr Thr Cys Ser
180 185 190
Ser Gly Ser Gly Ser Thr Thr Cys Ser Ser Gly Ser Gly Ser Thr Thr
195 200 205
Cys Ser Ser Gly Ser Gly Asp Ser Cys Asp Asp Met Pro Thr Ile Asp
210 215 220
Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His Phe
225 230 235 240
Cys Lys Gln Arg Gln Glu Lys Val Asn Ala Val Ile Lys Asn Cys Asn
245 250 255
Ser Cys Lys Glu Ser Gly Gly Thr Cys Asn Gly Glu Cys Lys Thr Glu
260 265 270
Cys Lys Asn Lys Cys Glu Ala Tyr Lys Thr Phe Ile Glu Glu Phe Cys
275 280 285
Thr Ala Asp Gly Gly Thr Ser Gly Ser Pro Trp Ser Lys Arg Trp Asp
290 295 300
Gln Ile Tyr Lys Met Tyr Ser Lys His Ile Glu Asp Ala Lys Arg Asn
305 310 315 320
Arg Lys Ala Gly Thr Lys Asn Cys Gly Pro Ser Ser Thr Thr Asn Val
325 330 335
Ser Val Ser Thr Asp Glu Asn Lys Cys Val Gln Ser
340 345
<210> 43
<211> 652
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 43
Asp Tyr Ile Lys Asp Asp Pro Tyr Phe Ala Glu Tyr Val Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Ser Ser Asp Ala Asn Asn Pro Ser Gly Glu Thr
20 25 30
Ala Asn His Asn Asp Glu Val Cys Asn Pro Asn Glu Ser Gly Ile Ala
35 40 45
Ser Val Glu Gln Ala Gln Thr Ser Asp Pro Ser Ser Asn Lys Thr Cys
50 55 60
Asn Thr His Ser Ser Ile Lys Ala Asn Lys Lys Lys Val Cys Lys His
65 70 75 80
Val Lys Leu Gly Val Arg Glu Asn Asp Lys Asp Leu Lys Ile Cys Val
85 90 95
Ile Glu His Thr Ser Leu Ser Gly Val Glu Asn Cys Cys Cys Gln Asp
100 105 110
Phe Leu Arg Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Ser Gly Ser
115 120 125
Ser Ser Asn Gly Ser Cys Asn Asn Lys Asn Gln Glu Ala Cys Glu Lys
130 135 140
Asn Leu Glu Lys Val Leu Ala Ser Leu Thr Asn Cys Tyr Lys Cys Asp
145 150 155 160
Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Lys Asn Trp Ile Trp
165 170 175
Lys Lys Ser Ser Gly Asn Glu Lys Gly Leu Gln Lys Glu Tyr Ala Asn
180 185 190
Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Cys Leu Val Val Cys
195 200 205
Leu Asp Glu Lys Glu Gly Lys Thr Gln Glu Leu Lys Asn Ile Arg Thr
210 215 220
Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu Gly
225 230 235 240
Lys Asn Leu Lys Lys Arg Tyr Pro Gln Asn Lys Asn Asp Asp Asn Asn
245 250 255
Ser Lys Leu Cys Lys Asp Leu Lys Tyr Ser Phe Ala Asp Tyr Gly Asp
260 265 270
Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp Leu
275 280 285
Glu Leu Asn Leu Gln Asn Asn Phe Gly Lys Leu Phe Arg Lys Tyr Ile
290 295 300
Lys Lys Asn Ile Ser Thr Glu Gln Asp Thr Leu Tyr Ser Ser Leu Asp
305 310 315 320
Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Leu
325 330 335
Ala Met Lys His Gly Ala Gly Met Asn Ser Thr Thr Cys Ser Ser Gly
340 345 350
Ser Gly Ser Thr Thr Cys Ser Ser Gly Ser Gly Ser Thr Thr Cys Ser
355 360 365
Ser Gly Ser Gly Asp Ser Cys Asp Asp Met Pro Thr Thr Asp Phe Ile
370 375 380
Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His Phe Cys Lys
385 390 395 400
Gln Arg Gln Glu Lys Val Asn Ala Val Ile Lys Asn Cys Asn Ser Cys
405 410 415
Lys Glu Ser Gly Gly Thr Cys Asn Gly Glu Cys Lys Thr Glu Cys Lys
420 425 430
Asn Lys Cys Glu Ala Tyr Lys Thr Phe Ile Glu Glu Phe Cys Thr Ala
435 440 445
Asp Gly Gly Thr Ser Gly Ser Pro Trp Ser Lys Arg Trp Asp Gln Ile
450 455 460
Tyr Lys Met Tyr Ser Lys His Ile Glu Asp Ala Lys Arg Asn Arg Lys
465 470 475 480
Ala Gly Thr Lys Asn Cys Gly Pro Ser Ser Thr Thr Asn Val Ser Val
485 490 495
Ser Thr Asp Glu Asn Lys Cys Val Gln Ser Asp Ile Asp Ser Phe Phe
500 505 510
Lys His Leu Ile Asp Ile Gly Leu Thr Thr Pro Ser Ser Tyr Leu Ser
515 520 525
Ile Val Leu Asp Asp Asn Ile Cys Gly Glu Asp Lys Ala Pro Trp Thr
530 535 540
Thr Tyr Thr Thr Tyr Thr Thr Thr Lys Lys Cys Asn Lys Glu Thr Asp
545 550 555 560
Lys Ser Lys Ser Gln Ser Cys Asn Thr Ala Val Val Val Asn Val Pro
565 570 575
Ser Pro Leu Gly Asn Thr Pro His Gly Tyr Lys Tyr Ala Cys Glu Cys
580 585 590
Lys Ile Pro Thr Thr Glu Glu Thr Cys Asp Asp Arg Lys Glu Tyr Met
595 600 605
Asn Gln Trp Ile Ile Asp Thr Ser Lys Lys Gln Lys Gly Ser Gly Ser
610 615 620
Gly Lys Asp Asp Tyr Glu Leu Tyr Thr Tyr Asn Gly Val Asp Val Lys
625 630 635 640
Pro Thr Thr Val Arg Ser Asn Ser Thr Lys Leu Asp
645 650
<210> 44
<211> 628
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 44
Asp Tyr Ile Lys Asp Asp Pro Tyr Ser Ala Gln Tyr Thr Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Pro Ser Asp Ala Asn Thr Ser Ser Glu Lys Ile
20 25 30
Gln Lys Asn Asn Asp Glu Ala Cys Asn Cys Asn Glu Ser Gly Ile Ser
35 40 45
Ser Val Gly Gln Ala Gln Thr Ser Gly Pro Ser Ser Asn Lys Thr Cys
50 55 60
Ile Thr His Ser Ser Ile Lys Ala Asn Lys Lys Lys Val Cys Lys Asp
65 70 75 80
Val Lys Leu Gly Ile Asn Asn Asn Asp Lys Val Leu Arg Val Cys Val
85 90 95
Ile Glu Asp Thr Ser Leu Ser Gly Val Asp Asn Cys Cys Cys Gln Asp
100 105 110
Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Arg Gly Ser
115 120 125
Ser Ser Asn Gly Ser Cys Asn Asn Asn Asn Glu Glu Ala Cys Glu Lys
130 135 140
Asn Leu Asp Glu Ala Pro Ala Ser Leu His Asn Gly Tyr Lys Asn Gln
145 150 155 160
Lys Cys Lys Ser Gly Thr Ser Arg Ser Lys Lys Lys Trp Ile Trp Lys
165 170 175
Lys Ser Ser Gly Asn Glu Lys Gly Leu Gln Glu Glu Tyr Ala Asn Thr
180 185 190
Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Cys Leu Val Cys Leu His
195 200 205
Glu Lys Glu Gly Lys Thr Gln His Lys Thr Ile Ser Thr Asn Ser Glu
210 215 220
Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu Gly Lys Asn Leu
225 230 235 240
Lys Thr Ser His Glu Lys Lys Asn Asp Asp Asn Gly Lys Lys Leu Cys
245 250 255
Lys Ala Leu Glu Tyr Ser Phe Ala Asp Tyr Gly Asp Leu Ile Lys Gly
260 265 270
Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp Leu Glu Leu Asn Leu
275 280 285
Gln Lys Ala Phe Gly Lys Leu Phe Arg Lys Tyr Ile Lys Lys Asn Asn
290 295 300
Thr Ala Glu Gln Asp Thr Ser Tyr Ser Ser Leu Asp Glu Leu Arg Glu
305 310 315 320
Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Ile Ala Met Lys His
325 330 335
Gly Ala Gly Met Asn Gly Thr Thr Cys Ser Cys Ser Gly Asp Ser Ser
340 345 350
Asn Asp Met Pro Thr Ile Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu
355 360 365
Gln Glu Trp Val Glu His Phe Cys Glu Gln Arg Gln Ala Lys Val Lys
370 375 380
Asp Val Ile Thr Asn Cys Lys Ser Cys Lys Glu Ser Gly Asn Lys Cys
385 390 395 400
Lys Thr Glu Cys Lys Thr Lys Cys Lys Asp Glu Cys Glu Lys Tyr Lys
405 410 415
Thr Phe Ile Glu Asp Cys Asn Gly Gly Gly Thr Gly Thr Ala Gly Ser
420 425 430
Ser Trp Val Lys Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys His
435 440 445
Ile Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly
450 455 460
Pro Ser Ser Ile Thr Asn Ala Ala Ala Ser Thr Asp Glu Asn Lys Cys
465 470 475 480
Val Gln Ser Asp Ile Asp Ser Phe Phe Lys His Leu Ile Asp Ile Gly
485 490 495
Leu Thr Thr Pro Ser Ser Tyr Leu Ser Asn Val Leu Asp Glu Asn Ser
500 505 510
Cys Gly Asp Asp Lys Ala Pro Trp Thr Thr Tyr Thr Thr Tyr Thr Thr
515 520 525
Thr Lys Asn Cys Asp Ile Gln Lys Asp Lys Ser Lys Ser Gln Pro Ile
530 535 540
Asn Thr Ser Val Val Val Asn Val Pro Ser Pro Leu Gly Asn Thr Pro
545 550 555 560
Tyr Arg Tyr Lys Tyr Ala Cys Glu Cys Lys Ile Pro Thr Thr Glu Glu
565 570 575
Ser Cys Asp Asp Arg Lys Glu Tyr Met Asn Gln Trp Ser Cys Gly Ser
580 585 590
Ala Arg Thr Met Lys Arg Gly Tyr Lys Asn Asp Asn Tyr Glu Leu Cys
595 600 605
Lys Tyr Asn Gly Val Asp Val Lys Pro Thr Thr Val Arg Ser Asn Ser
610 615 620
Ser Lys Leu Asp
625
<210> 45
<211> 653
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 45
Asp Tyr Ile Lys Gly Asp Pro Tyr Phe Ala Glu Tyr Ala Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Pro Ser Asp Thr Glu Asn Ala Ser Glu Thr Pro
20 25 30
Ser Lys Tyr Tyr Asp Glu Ala Cys Asn Pro Asn Glu Ser Glu Ile Ala
35 40 45
Ser Val Glu Gln Ala Gln Thr Ser Gly Pro Ser Ser Asn Lys Thr Cys
50 55 60
Ile Thr His Ser Ser Ile Lys Thr Asn Lys Lys Lys Glu Cys Lys Asp
65 70 75 80
Val Lys Leu Gly Val Arg Glu Asn Asp Lys Asp Leu Lys Ile Cys Val
85 90 95
Ile Glu Asp Thr Ser Leu Ser Gly Val Asp Asn Cys Cys Phe Lys Asp
100 105 110
Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Arg Gly Ser
115 120 125
Ser Ser Asn Asp Ser Cys Asn Asn Asn Asn Glu Glu Ala Cys Glu Lys
130 135 140
Asn Leu Asp Glu Ala Leu Ala Ser Leu Thr Asn Gly Tyr Lys Cys Asp
145 150 155 160
Lys Cys Lys Ser Gly Thr Ser Thr Val Asn Lys Lys Trp Thr Trp Arg
165 170 175
Lys Ser Ser Gly Asn Glu Glu Gly Leu Gln Lys Glu Tyr Ala Asn Thr
180 185 190
Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Cys Leu Val Cys Leu His
195 200 205
Glu Lys Glu Gly Lys Thr Lys His Lys Thr Ile Ser Thr Asn Ser Glu
210 215 220
Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu Gly Lys Asn Leu
225 230 235 240
Lys Thr Ser His Glu Lys Lys Asn Asp Asp Asn Gly Lys Lys Leu Cys
245 250 255
Lys Ala Leu Glu Tyr Ser Phe Ala Asp Tyr Gly Asp Leu Ile Lys Gly
260 265 270
Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp Leu Glu Leu Asn Leu
275 280 285
Gln Lys Ala Phe Gly Lys Leu Phe Arg Lys Tyr Ile Lys Lys Asn Asn
290 295 300
Thr Ala Glu Gln Asp Thr Ser Tyr Ser Ser Leu Asp Glu Leu Arg Glu
305 310 315 320
Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Thr Ala Met Lys His
325 330 335
Gly Ala Glu Met Asn Gly Thr Thr Cys Ser Ser Gly Ser Gly Asp Asn
340 345 350
Gly Asp Ser Ser Ile Thr Gly Ser Ser Asp Ser Gly Ser Thr Thr Cys
355 360 365
Ser Gly Asp Asn Gly Ser Ile Ser Cys Asp Asp Ile Pro Thr Thr Asp
370 375 380
Phe Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His Phe
385 390 395 400
Cys Glu Gln Arg Gln Ala Lys Val Lys Asp Val Ile Asn Ser Cys Asn
405 410 415
Ser Cys Asn Glu Ser Gly Gly Thr Cys Asn Gly Glu Cys Lys Thr Lys
420 425 430
Cys Lys Asp Glu Cys Glu Lys Tyr Lys Lys Phe Ile Glu Asp Cys Asn
435 440 445
Gly Gly Asp Gly Thr Ala Gly Ser Ser Trp Val Lys Arg Trp Asp Gln
450 455 460
Ile Tyr Lys Arg Tyr Ser Lys His Ile Glu Asp Ala Lys Arg Asn Arg
465 470 475 480
Lys Ala Gly Thr Lys Asn Cys Gly Pro Ser Ser Ile Thr Asn Ala Ala
485 490 495
Ala Ser Thr Asp Glu Asn Lys Cys Val Gln Ser Asp Val Asp Ser Phe
500 505 510
Phe Lys His Leu Ile Asp Ile Gly Leu Thr Thr Pro Ser Ser Tyr Leu
515 520 525
Ser Ile Val Leu Asp Glu Asn Ser Cys Gly Asp Asp Lys Ala Pro Trp
530 535 540
Thr Thr Tyr Thr Thr Tyr Thr Thr Thr Glu Lys Cys Asn Lys Glu Arg
545 550 555 560
Asp Lys Ser Lys Ser Gln Ser Ser Asp Thr Leu Val Val Val Asn Val
565 570 575
Pro Ser Pro Leu Gly Asn Thr Pro His Glu Tyr Lys Tyr Ala Cys Glu
580 585 590
Cys Lys Ile Pro Thr Asn Glu Glu Thr Cys Asp Asp Arg Lys Asp Tyr
595 600 605
Met Asn Gln Trp Ile Ser Asp Thr Ser Lys Lys Gln Lys Gly Ser Gly
610 615 620
Ser Gly Lys Asp Tyr Tyr Glu Leu Tyr Thr Tyr Asn Gly Val Gln Ile
625 630 635 640
Lys Gln Ala Ala Gly Arg Ser Ser Ser Thr Lys Leu Asp
645 650
<210> 46
<211> 490
<212> PRT
<213> 恶性疟原虫
<400> 46
Lys Cys Asp Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Asn Lys
1 5 10 15
Trp Ile Trp Lys Lys Tyr Ser Gly Asn Gly Glu Gly Leu Gln Lys Glu
20 25 30
Tyr Ala Asn Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Cys Leu
35 40 45
Val Cys Leu His Glu Lys Glu Gly Lys Thr Gln His Lys Thr Ile Ser
50 55 60
Thr Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu
65 70 75 80
Gly Lys Asn Leu Lys Lys Arg Tyr Pro Gln Asn Lys Asn Asp Asp Asn
85 90 95
Asn Ser Lys Leu Cys Lys Ala Leu Glu Tyr Ser Phe Ala Asp Tyr Gly
100 105 110
Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp
115 120 125
Leu Glu Leu Asn Leu Gln Lys Ala Phe Gly Lys Leu Phe Arg Lys Tyr
130 135 140
Ile Lys Lys Asn Asn Thr Ala Glu Gln Asp Thr Ser Tyr Ser Ser Leu
145 150 155 160
Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp
165 170 175
Thr Ala Met Lys His Gly Ala Glu Met Asn Gly Thr Thr Cys Ser Ser
180 185 190
Gly Ser Gly Asp Asn Gly Asp Ser Ser Cys Asp Asp Ile Pro Thr Ile
195 200 205
Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His
210 215 220
Phe Cys Lys Gln Arg Gln Ala Lys Val Lys Asp Val Ile Asn Ser Cys
225 230 235 240
Asn Ser Cys Lys Asn Thr Ser Gly Glu Arg Lys Ile Gly Gly Thr Cys
245 250 255
Asn Ser Asp Cys Glu Lys Lys Cys Lys Val Ala Cys Asp Ala Tyr Lys
260 265 270
Thr Phe Ile Glu Glu Cys Arg Thr Ala Val Gly Gly Thr Ala Gly Ser
275 280 285
Ser Trp Val Lys Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys His
290 295 300
Ile Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly
305 310 315 320
Pro Ser Ser Thr Thr Asn Ala Ala Glu Asn Lys Cys Val Gln Ser Asp
325 330 335
Ile Asp Ser Phe Phe Lys His Leu Ile Asp Ile Gly Leu Thr Thr Pro
340 345 350
Ser Ser Tyr Leu Ser Asn Val Leu Asp Glu Asn Ser Cys Gly Ala Asp
355 360 365
Lys Ala Pro Trp Thr Thr Tyr Thr Thr Tyr Thr Thr Tyr Thr Thr Tyr
370 375 380
Thr Thr Tyr Thr Thr Thr Glu Lys Cys Asn Lys Glu Arg Asp Lys Ser
385 390 395 400
Lys Ser Gln Gln Ser Asn Thr Ser Val Val Val Asn Val Pro Ser Pro
405 410 415
Leu Gly Asn Thr Pro His Glu Tyr Lys Tyr Ala Cys Glu Cys Lys Ile
420 425 430
Pro Thr Thr Glu Glu Thr Cys Asp Asp Arg Lys Glu Tyr Met Asn Gln
435 440 445
Trp Ile Ile Asp Asn Thr Lys Asn Pro Lys Gly Ser Gly Ser Thr Asp
450 455 460
Asn Asp Tyr Glu Leu Tyr Thr Tyr Asn Gly Val Gln Ile Lys Gln Ala
465 470 475 480
Ala Gly Arg Ser Ser Ser Thr Lys Leu Asp
485 490
<210> 47
<211> 335
<212> PRT
<213> 恶性疟原虫
<400> 47
Lys Cys Glu Lys Cys Lys Ser Gly Thr Ser Thr Val Asn Asn Lys Trp
1 5 10 15
Ile Trp Arg Lys Ser Ser Gly Lys Glu Gly Gly Leu Gln Lys Glu Tyr
20 25 30
Ala Asn Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu Gly
35 40 45
Asn Leu Pro Lys Leu Glu Asn Val Cys Lys Gly Val Thr Asp Ile Ile
50 55 60
Tyr Asp Thr Lys Glu Lys Phe Leu Ser Gly Cys Leu Ile Ala Ala Phe
65 70 75 80
His Glu Gly Lys Asn Leu Lys Thr Thr Tyr Leu Glu Lys Lys Asn Asp
85 90 95
Asp Asn Gly Lys Lys Leu Cys Lys Ala Leu Glu Tyr Ser Phe Ala Asp
100 105 110
Tyr Gly Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr
115 120 125
Lys Asp Leu Glu Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe Arg
130 135 140
Lys Tyr Ile Lys Lys Asn Asn Thr Ala Glu Gln Asp Thr Ser Tyr Ser
145 150 155 160
Ser Leu Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr
165 170 175
Ile Trp Ile Ala Met Lys His Gly Ala Gly Met Asn Gly Thr Thr Cys
180 185 190
Ser Ser Gly Ser Gly Asp Ser Ser Asn Asp Ile Pro Thr Thr Asp Phe
195 200 205
Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu Asn Phe Cys
210 215 220
Glu Gln Arg Gln Ala Lys Val Lys Pro Val Ile Glu Asn Cys Asn Ser
225 230 235 240
Cys Lys Glu Ser Gly Gly Thr Cys Asn Gly Glu Cys Lys Thr Lys Cys
245 250 255
Lys Val Ala Cys Asp Ala Tyr Lys Lys Phe Ile Asp Gly Thr Gly Ser
260 265 270
Gly Gly Gly Ser Arg Pro Thr Gly Ile Ala Gly Ser Ser Trp Ser Lys
275 280 285
Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys His Ile Glu Asp Ala
290 295 300
Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Pro Ser Ser Ile
305 310 315 320
Thr Asn Val Ser Val Ser Thr Asp Glu Asn Lys Cys Val Gln Ser
325 330 335
<210> 48
<211> 637
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 48
Asn Tyr Ile Lys Asp Asp Pro Tyr Ser Lys Glu Tyr Val Thr Lys Leu
1 5 10 15
Ser Phe Ile Pro Asn Ser Ser Asp Ala Asn Thr Ser Ser Glu Lys Ile
20 25 30
Gln Lys Asn Asn Asp Glu Val Cys Asn Pro Asn Glu Ser Gly Ile Ser
35 40 45
Ser Val Glu Gln Ala Gln Thr Ser Asp Pro Ser Ser Asn Lys Thr Cys
50 55 60
Ile Thr His Ser Ser Ile Lys Ala Asn Lys Lys Lys Glu Cys Lys Asp
65 70 75 80
Val Lys Leu Gly Val Arg Glu Asn Asp Lys Asp Leu Lys Ile Cys Val
85 90 95
Ile Glu His Thr Ser Leu Ser Gly Val Asp Asn Cys Cys Phe Lys Asp
100 105 110
Phe Leu Arg Met Leu Gln Glu Pro Arg Ile Asp Lys Asn Gln Arg Gly
115 120 125
Ser Ser Ser Asn Gly Ser Cys Asp Lys Asn Ser Glu Glu Ala Cys Glu
130 135 140
Lys Asn Leu Asp Glu Ala Leu Ala Ser Leu Thr Asn Gly Tyr Lys Cys
145 150 155 160
Asp Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Asn Lys Trp Ile
165 170 175
Trp Lys Lys Phe Pro Gly Lys Glu Gly Gly Leu Gln Glu Glu Tyr Ala
180 185 190
Asn Thr Ile Gly Leu Pro Pro Arg Thr Gln Tyr Leu Cys Leu Val Val
195 200 205
Cys Leu Asp Glu Lys Glu Gly Lys Thr Gln Glu Leu Lys Asn Ile Arg
210 215 220
Thr Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu
225 230 235 240
Gly Lys Asn Leu Lys Thr Thr Tyr Pro Gln Lys Lys Asn Asp Asp Asn
245 250 255
Gly Lys Lys Leu Cys Lys Asp Leu Lys Tyr Ser Phe Ala Asp Tyr Gly
260 265 270
Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asn
275 280 285
Val Glu Leu Asn Leu Gln Asn Asn Phe Gly Lys Leu Phe Arg Lys Tyr
290 295 300
Ile Lys Lys Asn Asn Thr Ala Glu Gln Asp Thr Ser Tyr Ser Ser Leu
305 310 315 320
Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp
325 330 335
Leu Ala Met Lys His Gly Ala Glu Met Asn Ser Thr Thr Cys Cys Gly
340 345 350
Asp Gly Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp Ile Pro Thr
355 360 365
Ile Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu
370 375 380
His Phe Cys Lys Gln Arg Gln Ala Lys Val Lys Asp Val Ile Thr Asn
385 390 395 400
Cys Asn Ser Cys Lys Glu Ser Gly Asn Lys Cys Lys Thr Glu Cys Lys
405 410 415
Asn Lys Cys Lys Asp Glu Cys Glu Lys Tyr Lys Lys Phe Ile Glu Ala
420 425 430
Cys Gly Thr Ala Val Gly Gly Thr Gly Thr Ala Gly Ser Pro Trp Ser
435 440 445
Lys Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys His Ile Glu Asp
450 455 460
Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Pro Ser Ser
465 470 475 480
Thr Thr Asn Ala Ala Glu Asn Lys Cys Val Gln Ser Asp Ile Asp Ser
485 490 495
Phe Phe Lys His Leu Ile Asp Ile Gly Leu Thr Thr Pro Ser Ser Tyr
500 505 510
Leu Ser Ile Val Leu Asp Asp Asn Ile Cys Gly Ala Asp Lys Ala Pro
515 520 525
Trp Thr Thr Tyr Thr Thr Tyr Thr Thr Glu Asn Cys Asp Ile Gln Lys
530 535 540
Lys Thr Pro Lys Ser Gln Ser Cys Asp Thr Leu Val Val Val Asn Val
545 550 555 560
Pro Ser Pro Leu Gly Asn Thr Pro His Gly Tyr Lys Tyr Ala Cys Gln
565 570 575
Cys Arg Thr Pro Asn Lys Gln Glu Ser Cys Asp Asp Arg Lys Glu Tyr
580 585 590
Met Asn Gln Trp Ile Ile Asp Asn Thr Lys Asn Pro Lys Gly Ser Gly
595 600 605
Ser Gly Lys Asp Tyr Tyr Glu Leu Cys Lys Tyr Asn Gly Val Lys Glu
610 615 620
Thr Lys Pro Leu Gly Thr Leu Lys Asn Ser Lys Leu Asp
625 630 635
<210> 49
<211> 330
<212> PRT
<213> 恶性疟原虫
<400> 49
Lys Cys Asp Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Asn Lys
1 5 10 15
Trp Ile Trp Arg Lys Phe Pro Gly Lys Glu Gly Gly Leu Gln Lys Glu
20 25 30
Tyr Ala Asn Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Cys Leu
35 40 45
Val Cys Leu His Glu Lys Glu Gly Lys Thr Gln His Lys Thr Ile Ser
50 55 60
Thr Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu
65 70 75 80
Gly Lys Asn Leu Lys Thr Thr Tyr Leu Glu Lys Lys Asn Ala Glu Asn
85 90 95
Lys Lys Lys Leu Cys Lys Ala Leu Lys Tyr Ser Phe Ala Asp Tyr Gly
100 105 110
Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp
115 120 125
Leu Glu Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe Arg Lys Tyr
130 135 140
Ile Lys Lys Asn Asn Thr Ala Glu Gln Asp Thr Ser Tyr Ser Ser Leu
145 150 155 160
Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp
165 170 175
Thr Ala Met Lys His Gly Ala Gly Met Asn Gly Thr Met Cys Asn Ala
180 185 190
Asp Gly Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp Met Pro Thr
195 200 205
Thr Asp Phe Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu
210 215 220
His Phe Cys Lys Gln Arg Gln Ala Lys Val Lys Asp Val Ile Glu Asn
225 230 235 240
Cys Lys Ser Cys Lys Glu Ser Gly Asn Lys Cys Lys Thr Glu Cys Lys
245 250 255
Asn Lys Cys Asp Ala Tyr Lys Thr Phe Ile Glu Glu Cys Gly Thr Ala
260 265 270
Val Gly Gly Thr Ala Gly Ser Ser Trp Val Lys Arg Trp Asp Gln Ile
275 280 285
Tyr Lys Arg Tyr Ser Lys His Ile Glu Asp Ala Lys Arg Asn Arg Lys
290 295 300
Ala Gly Thr Lys Asn Cys Gly Thr Ser Ser Thr Thr Asn Ala Ala Ala
305 310 315 320
Ser Thr Ala Glu Asn Lys Cys Val Gln Ser
325 330
<210> 50
<211> 269
<212> PRT
<213> 恶性疟原虫
<400> 50
Asn Tyr Ile Lys Asp Asp Pro Tyr Ser Lys Glu Tyr Val Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Ser Ser Asp Ala Glu Asn Ala Ser Glu Thr Pro
20 25 30
Ser Lys Tyr Tyr Asp Glu Ala Cys Asn Cys Asn Glu Ser Gly Ile Ser
35 40 45
Ser Val Glu Gln Ala Ser Ile Ser Asp Arg Ser Ser Gln Lys Ala Cys
50 55 60
Asn Thr His Ser Phe Ile Gly Ala Asn Lys Lys Lys Val Cys Lys His
65 70 75 80
Val Lys Leu Gly Val Arg Glu Asn Asp Lys Asp Leu Lys Ile Cys Val
85 90 95
Ile Glu Asp Asp Ser Leu Arg Gly Val Glu Asn Cys Cys Phe Lys Asp
100 105 110
Phe Leu Arg Met Leu Gln Glu Pro Arg Ile Asp Lys Asn Gln Arg Gly
115 120 125
Ser Ser Ser Asn Asp Ser Cys Asn Asn Asn Asn Glu Glu Ala Cys Glu
130 135 140
Lys Asn Leu Asp Glu Ala Leu Ala Ser Leu His Asn Gly Tyr Lys Asn
145 150 155 160
Gln Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Asn Lys Trp Ile
165 170 175
Trp Lys Lys Ser Ser Gly Lys Glu Gly Gly Leu Gln Lys Glu Tyr Ala
180 185 190
Asn Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Cys Leu Val Cys
195 200 205
Leu His Glu Lys Glu Gly Lys Thr Gln His Lys Thr Ile Ser Thr Asn
210 215 220
Ser Glu Leu Leu Lys Glu Trp Ile Ile Asp Ala Phe His Glu Gly Lys
225 230 235 240
Asn Leu Lys Thr Thr Tyr Leu Glu Lys Lys Lys Gly Asp Asn Gly Lys
245 250 255
Lys Leu Cys Lys Ala Leu Lys Tyr Ser Phe Ala Asp Tyr
260 265
<210> 51
<211> 347
<212> PRT
<213> 恶性疟原虫
<400> 51
Lys Cys Asp Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Lys Asn
1 5 10 15
Trp Ile Trp Lys Lys Ser Ser Gly Lys Glu Gly Gly Leu Gln Lys Glu
20 25 30
Tyr Ala Asn Thr Ile Ala Leu Pro Pro Arg Thr Gln Ser Leu Cys Leu
35 40 45
Val Val Cys Leu His Glu Lys Glu Gly Lys Thr Gln His Lys Thr Ile
50 55 60
Ser Thr Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Asp Ala Phe His
65 70 75 80
Glu Gly Lys Asn Leu Lys Thr Thr Tyr Leu Glu Lys Gln Asn Ala Asp
85 90 95
Asn Gly Lys Lys Asn Ala Asp Asn Asn Ser Lys Leu Cys Lys Asp Leu
100 105 110
Lys Tyr Ser Phe Ala Asp Tyr Gly Asp Leu Ile Lys Gly Thr Ser Ile
115 120 125
Trp Asp Asn Glu Tyr Thr Lys Asp Leu Glu Leu Asn Leu Gln Gln Ile
130 135 140
Phe Gly Lys Leu Phe Arg Lys Tyr Ile Lys Lys Asn Ile Ala Ser Asp
145 150 155 160
Glu Asn Thr Leu Tyr Ser Ser Leu Asp Glu Leu Arg Glu Ser Trp Trp
165 170 175
Asn Thr Asn Lys Lys Tyr Ile Trp Thr Ala Met Lys His Gly Ala Glu
180 185 190
Met Asn Gly Thr Thr Cys Ser Ser Gly Ser Gly Asp Ser Ser Ser Gly
195 200 205
Glu Asn Gln Thr Asn Ser Cys Asp Asp Ile Pro Thr Ile Asp Leu Ile
210 215 220
Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His Phe Cys Glu
225 230 235 240
Gln Arg Gln Ala Lys Val Lys Asp Val Ile Thr Asn Cys Lys Ser Cys
245 250 255
Lys Glu Ser Gly Gly Thr Cys Asn Ser Asp Cys Lys Thr Lys Cys Lys
260 265 270
Gly Glu Cys Glu Lys Tyr Lys Lys Phe Ile Glu Lys Cys Lys Gly Gly
275 280 285
Gly Thr Glu Gly Thr Ser Gly Ser Ser Trp Val Lys Arg Trp Tyr Gln
290 295 300
Ile Tyr Met Arg Tyr Ser Lys Tyr Ile Glu Asp Ala Lys Arg Asn Arg
305 310 315 320
Lys Ala Gly Thr Lys Ser Cys Gly Thr Ser Ser Gly Ala Asn Ser Gly
325 330 335
Val Thr Thr Thr Glu Ser Lys Cys Val Gln Ser
340 345
<210> 52
<211> 269
<212> PRT
<213> 恶性疟原虫
<400> 52
Asp Tyr Ile Lys Asp Asp Pro Tyr Ser Lys Glu Tyr Thr Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Ser Ser Asp Ala Asn Thr Ser Ser Glu Lys Ile
20 25 30
Gln Lys Asn Asn Asp Glu Val Cys Asn Pro Asn Glu Ser Glu Ile Ser
35 40 45
Ser Val Glu Gln Ala Gln Thr Ser Arg Pro Ser Ser Asn Lys Thr Cys
50 55 60
Ile Thr His Ser Ser Ile Lys Ala Asn Lys Lys Lys Val Cys Lys Asp
65 70 75 80
Val Lys Leu Gly Val Arg Glu Asn Asp Lys Val Leu Arg Val Cys Val
85 90 95
Ile Glu His Thr Ser Leu Ser Gly Val Glu Asn Cys Cys Cys Gln Asp
100 105 110
Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Arg Gly Ser
115 120 125
Ser Ser Asn Gly Ser Cys Asp Lys Asn Ser Glu Glu Ala Cys Glu Lys
130 135 140
Asn Leu Asp Glu Ala Leu Ala Ser Leu Thr Asn Cys Tyr Lys Asn Gln
145 150 155 160
Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Asn Lys Trp Ile Trp
165 170 175
Lys Lys Ser Ser Gly Asn Glu Lys Gly Leu Gln Lys Glu Tyr Ala Asn
180 185 190
Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Cys Leu Val Cys Leu
195 200 205
His Glu Lys Glu Gly Lys Thr Gln Glu Leu Lys Asn Ile Ser Thr Asn
210 215 220
Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu Gly Lys
225 230 235 240
Asn Leu Lys Thr Thr Tyr Pro Gln Asn Lys Asn Asp Asp Asn Gly Lys
245 250 255
Lys Leu Phe Lys Asp Leu Lys Tyr Ser Phe Ala Asp Tyr
260 265
<210> 53
<211> 646
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 53
Asp Tyr Ile Lys Asp Asp Pro Tyr Ser Lys Glu Tyr Thr Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Ser Ser Asp Ala Asn Thr Ser Ser Glu Lys Ile
20 25 30
Gln Lys Asn Asn Asp Glu Val Cys Asn Pro Asn Glu Ser Glu Ile Ser
35 40 45
Ser Val Glu Gln Ala Gln Thr Ser Arg Pro Ser Ser Asn Lys Thr Cys
50 55 60
Ile Thr His Ser Ser Ile Lys Ala Asn Lys Lys Lys Val Cys Lys Asp
65 70 75 80
Val Lys Leu Gly Val Arg Glu Asn Asp Lys Val Leu Arg Val Cys Val
85 90 95
Ile Glu His Thr Ser Leu Ser Gly Val Glu Asn Cys Cys Cys Gln Asp
100 105 110
Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Arg Gly Ser
115 120 125
Ser Ser Asn Gly Ser Cys Asp Lys Asn Ser Glu Glu Ala Cys Glu Lys
130 135 140
Asn Leu Asp Glu Ala Leu Ala Ser Leu Thr Asn Cys Tyr Lys Asn Gln
145 150 155 160
Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Asn Lys Trp Ile Trp
165 170 175
Lys Lys Ser Ser Gly Lys Glu Gly Gly Leu Gln Lys Glu Tyr Ala Asn
180 185 190
Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu Gly Asn Leu
195 200 205
Pro Lys Leu Glu Asn Val Cys Lys Gly Val Thr Asp Ile Asn Phe Asp
210 215 220
Thr Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Ala Ala Phe His Glu
225 230 235 240
Gly Lys Asn Leu Lys Thr Thr Tyr Leu Glu Lys Lys Asn Asp Asp Asn
245 250 255
Gly Lys Lys Leu Cys Lys Ala Leu Glu Tyr Ser Phe Ala Asp Tyr Gly
260 265 270
Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp
275 280 285
Leu Glu Leu Asn Leu Gln Lys Ala Phe Gly Lys Leu Phe Arg Lys Tyr
290 295 300
Ile Lys Lys Asn Asn Thr Ala Glu Gln Asp Thr Ser Tyr Ser Ser Leu
305 310 315 320
Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp
325 330 335
Thr Ala Met Lys His Gly Ala Gly Met Asn Gly Thr Thr Cys Ser Ser
340 345 350
Gly Ser Gly Asp Ser Ser Asn Asp Ile Pro Thr Thr Asp Phe Ile Pro
355 360 365
Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu Asn Phe Cys Glu Gln
370 375 380
Arg Gln Ala Lys Val Lys Asp Val Ile Glu Asn Cys Asn Ser Cys Lys
385 390 395 400
Asn Thr Ser Gly Glu Arg Lys Ile Gly Asp Thr Cys Asn Ser Asp Cys
405 410 415
Glu Lys Lys Cys Lys Asp Glu Cys Glu Lys Tyr Lys Lys Phe Ile Glu
420 425 430
Asp Cys Lys Gly Gly Asp Gly Thr Ala Gly Ser Ser Trp Val Lys Arg
435 440 445
Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys His Ile Glu Asp Ala Lys
450 455 460
Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Ile Thr Thr Gly Thr
465 470 475 480
Ile Ser Gly Glu Ser Ser Gly Ala Thr Ser Gly Val Thr Thr Thr Glu
485 490 495
Asn Lys Cys Val Gln Ser Asp Ile Asp Ser Phe Phe Lys His Leu Ile
500 505 510
Asp Ile Gly Leu Thr Thr Pro Ser Ser Tyr Leu Ser Asn Val Leu Asp
515 520 525
Asp Asn Ile Cys Gly Glu Asp Asn Ala Pro Trp Thr Thr Tyr Thr Thr
530 535 540
Tyr Thr Thr Glu Lys Cys Asn Lys Glu Thr Asp Lys Ser Lys Ser Gln
545 550 555 560
Gln Ser Asn Thr Ala Val Val Val Asn Val Pro Ser Pro Leu Gly Asn
565 570 575
Thr Pro His Gly Tyr Lys Tyr Ala Cys Glu Cys Lys Ile Pro Thr Thr
580 585 590
Glu Glu Thr Cys Asp Asp Arg Lys Glu Tyr Met Asn Gln Trp Ser Cys
595 600 605
Gly Ser Ala Gln Thr Val Arg Asp Arg Ser Gly Lys Asp Asp Tyr Glu
610 615 620
Leu Cys Lys Tyr Asn Gly Val Gln Ile Lys Gln Ala Ala Gly Thr Leu
625 630 635 640
Lys Asn Ser Lys Leu Asp
645
<210> 54
<211> 632
<212> PRT
<213> 恶性疟原虫
<400> 54
Asn Tyr Ile Lys Gly Asp Pro Tyr Phe Ala Glu Tyr Ala Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Ser Ser Asp Ala Asn Asn Pro Ser Glu Lys Ile
20 25 30
Gln Lys Asn Asn Asp Glu Val Cys Asn Cys Asn Glu Ser Gly Ile Ala
35 40 45
Ser Val Glu Gln Glu Gln Ile Ser Asp Pro Ser Ser Asn Lys Thr Cys
50 55 60
Ile Thr His Ser Ser Ile Lys Ala Asn Lys Lys Lys Val Cys Lys His
65 70 75 80
Val Lys Leu Gly Val Arg Glu Asn Asp Lys Asp Leu Arg Val Cys Val
85 90 95
Ile Glu His Thr Ser Leu Ser Gly Val Glu Asn Cys Cys Cys Gln Asp
100 105 110
Phe Leu Arg Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Ser Gly Ser
115 120 125
Ser Ser Asn Gly Ser Cys Asn Asn Lys Asn Gln Glu Ala Cys Glu Lys
130 135 140
Asn Leu Glu Lys Val Leu Ala Ser Leu Thr Asn Cys Tyr Lys Cys Asp
145 150 155 160
Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Lys Asn Trp Ile Trp
165 170 175
Lys Lys Ser Ser Gly Lys Glu Gly Gly Leu Gln Lys Glu Tyr Ala Asn
180 185 190
Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Cys Leu Val Val Cys
195 200 205
Leu Asp Glu Lys Gly Lys Lys Thr Gln Glu Leu Lys Asn Ile Arg Thr
210 215 220
Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu Gly
225 230 235 240
Lys Asn Leu Lys Pro Ser His Glu Lys Lys Asn Asp Asp Asn Gly Lys
245 250 255
Lys Leu Cys Lys Ala Leu Glu Tyr Ser Phe Ala Asp Tyr Gly Asp Leu
260 265 270
Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp Leu Glu
275 280 285
Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe Arg Lys Tyr Ile Lys
290 295 300
Lys Asn Asn Thr Ala Glu Gln Asp Thr Ser Tyr Ser Ser Leu Asp Glu
305 310 315 320
Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Leu Ala
325 330 335
Met Lys His Gly Ala Gly Met Asn Ser Thr Thr Cys Cys Gly Asp Gly
340 345 350
Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp Ile Pro Thr Ile Asp
355 360 365
Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His Phe
370 375 380
Cys Lys Gln Arg Gln Glu Lys Val Lys Pro Val Ile Glu Asn Cys Lys
385 390 395 400
Ser Cys Lys Glu Ser Gly Gly Thr Cys Asn Gly Glu Cys Lys Thr Glu
405 410 415
Cys Lys Asn Lys Cys Glu Val Tyr Lys Lys Phe Ile Glu Asp Cys Lys
420 425 430
Gly Gly Asp Gly Thr Ala Gly Ser Ser Trp Val Lys Arg Trp Asp Gln
435 440 445
Ile Tyr Lys Arg Tyr Ser Lys Tyr Ile Glu Asp Ala Lys Arg Asn Arg
450 455 460
Lys Ala Gly Thr Lys Asn Cys Gly Pro Ser Ser Thr Thr Asn Ala Ala
465 470 475 480
Glu Asn Lys Cys Val Gln Ser Asp Ile Asp Ser Phe Phe Lys His Leu
485 490 495
Ile Asp Ile Gly Leu Thr Thr Pro Ser Ser Tyr Leu Ser Ile Val Leu
500 505 510
Asp Asp Asn Ile Cys Gly Ala Asp Lys Ala Pro Trp Thr Thr Tyr Thr
515 520 525
Thr Tyr Thr Thr Thr Glu Lys Cys Asn Lys Glu Thr Asp Lys Ser Lys
530 535 540
Leu Gln Gln Cys Asn Thr Ala Val Val Val Asn Val Pro Ser Pro Leu
545 550 555 560
Gly Asn Thr Pro His Gly Tyr Lys Tyr Ala Cys Gln Cys Lys Ile Pro
565 570 575
Thr Asn Glu Glu Thr Cys Asp Asp Arg Lys Glu Tyr Met Asn Gln Trp
580 585 590
Ser Cys Gly Ser Ala Arg Thr Met Lys Arg Gly Tyr Lys Asn Asp Asn
595 600 605
Tyr Glu Leu Cys Lys Tyr Asn Gly Val Asp Val Lys Pro Thr Thr Val
610 615 620
Arg Ser Asn Ser Ser Lys Leu Asp
625 630
<210> 55
<211> 2730
<212> PRT
<213> 恶性疟原虫
<400> 55
Met Asp Lys Ser Ser Ile Ala Asn Lys Ile Glu Ala Tyr Leu Gly Ala
1 5 10 15
Lys Ser Asp Asp Ser Lys Ile Asp Gln Ser Leu Lys Ala Asp Pro Ser
20 25 30
Glu Val Gln Tyr Tyr Gly Ser Gly Gly Asp Gly Tyr Tyr Leu Arg Lys
35 40 45
Asn Ile Cys Lys Ile Thr Val Asn His Ser Asp Ser Gly Thr Asn Asp
50 55 60
Pro Cys Asp Arg Ile Pro Pro Pro Tyr Gly Asp Asn Asp Gln Trp Lys
65 70 75 80
Cys Ala Ile Ile Leu Ser Lys Val Ser Glu Lys Pro Glu Asn Val Phe
85 90 95
Val Pro Pro Arg Arg Gln Arg Met Cys Ile Asn Asn Leu Glu Lys Leu
100 105 110
Asn Val Asp Lys Ile Arg Asp Lys His Ala Phe Leu Ala Asp Val Leu
115 120 125
Leu Thr Ala Arg Asn Glu Gly Glu Arg Ile Val Gln Asn His Pro Asp
130 135 140
Thr Asn Ser Ser Asn Val Cys Asn Ala Leu Glu Arg Ser Phe Ala Asp
145 150 155 160
Ile Ala Asp Ile Ile Arg Gly Thr Asp Leu Trp Lys Gly Thr Asn Ser
165 170 175
Asn Leu Glu Gln Asn Leu Lys Gln Met Phe Ala Lys Ile Arg Glu Asn
180 185 190
Asp Lys Val Leu Gln Asp Lys Tyr Pro Lys Asp Gln Asn Tyr Arg Lys
195 200 205
Leu Arg Glu Asp Trp Trp Asn Ala Asn Arg Gln Lys Val Trp Glu Val
210 215 220
Ile Thr Cys Gly Ala Arg Ser Asn Asp Leu Leu Ile Lys Arg Gly Trp
225 230 235 240
Arg Thr Ser Gly Lys Ser Asn Gly Asp Asn Lys Leu Glu Leu Cys Arg
245 250 255
Lys Cys Gly His Tyr Glu Glu Lys Val Pro Thr Lys Leu Asp Tyr Val
260 265 270
Pro Gln Phe Leu Arg Trp Leu Thr Glu Trp Ile Glu Asp Phe Tyr Arg
275 280 285
Glu Lys Gln Asn Leu Ile Asp Asp Met Glu Arg His Arg Glu Glu Cys
290 295 300
Thr Ser Glu Asp His Lys Ser Lys Glu Gly Thr Ser Tyr Cys Ser Thr
305 310 315 320
Cys Lys Asp Lys Cys Lys Lys Tyr Cys Glu Cys Val Lys Lys Trp Lys
325 330 335
Ser Glu Trp Glu Asn Gln Lys Asn Lys Tyr Thr Glu Leu Tyr Gln Gln
340 345 350
Asn Lys Asn Glu Thr Ser Gln Lys Asn Thr Ser Arg Tyr Asp Asp Tyr
355 360 365
Val Lys Asp Phe Phe Lys Lys Leu Glu Ala Asn Tyr Ser Ser Leu Glu
370 375 380
Asn Tyr Ile Lys Gly Asp Pro Tyr Phe Ala Glu Tyr Ala Thr Lys Leu
385 390 395 400
Ser Phe Ile Leu Asn Ser Ser Asp Ala Asn Asn Pro Ser Glu Lys Ile
405 410 415
Gln Lys Asn Asn Asp Glu Val Cys Asn Cys Asn Glu Ser Gly Ile Ala
420 425 430
Ser Val Glu Gln Glu Gln Ile Ser Asp Pro Ser Ser Asn Lys Thr Cys
435 440 445
Ile Thr His Ser Ser Ile Lys Ala Asn Lys Lys Lys Val Cys Lys His
450 455 460
Val Lys Leu Gly Val Arg Glu Asn Asp Lys Asp Leu Arg Val Cys Val
465 470 475 480
Ile Glu His Thr Ser Leu Ser Gly Val Glu Asn Cys Cys Cys Gln Asp
485 490 495
Phe Leu Arg Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Ser Gly Ser
500 505 510
Ser Ser Asn Gly Ser Cys Asn Asn Lys Asn Gln Glu Ala Cys Glu Lys
515 520 525
Asn Leu Glu Lys Val Leu Ala Ser Leu Thr Asn Cys Tyr Lys Cys Asp
530 535 540
Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Lys Asn Trp Ile Trp
545 550 555 560
Lys Lys Ser Ser Gly Lys Glu Gly Gly Leu Gln Lys Glu Tyr Ala Asn
565 570 575
Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Cys Leu Val Val Cys
580 585 590
Leu Asp Glu Lys Gly Lys Lys Thr Gln Glu Leu Lys Asn Ile Arg Thr
595 600 605
Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu Gly
610 615 620
Lys Asn Leu Lys Pro Ser His Glu Lys Lys Asn Asp Asp Asn Gly Lys
625 630 635 640
Lys Leu Cys Lys Ala Leu Glu Tyr Ser Phe Ala Asp Tyr Gly Asp Leu
645 650 655
Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp Leu Glu
660 665 670
Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe Arg Lys Tyr Ile Lys
675 680 685
Lys Asn Asn Thr Ala Glu Gln Asp Thr Ser Tyr Ser Ser Leu Asp Glu
690 695 700
Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Leu Ala
705 710 715 720
Met Lys His Gly Ala Gly Met Asn Ser Thr Thr Cys Cys Gly Asp Gly
725 730 735
Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp Ile Pro Thr Ile Asp
740 745 750
Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His Phe
755 760 765
Cys Lys Gln Arg Gln Glu Lys Val Lys Pro Val Ile Glu Asn Cys Lys
770 775 780
Ser Cys Lys Glu Ser Gly Gly Thr Cys Asn Gly Glu Cys Lys Thr Glu
785 790 795 800
Cys Lys Asn Lys Cys Glu Val Tyr Lys Lys Phe Ile Glu Asp Cys Lys
805 810 815
Gly Gly Asp Gly Thr Ala Gly Ser Ser Trp Val Lys Arg Trp Asp Gln
820 825 830
Ile Tyr Lys Arg Tyr Ser Lys Tyr Ile Glu Asp Ala Lys Arg Asn Arg
835 840 845
Lys Ala Gly Thr Lys Asn Cys Gly Pro Ser Ser Thr Thr Asn Ala Ala
850 855 860
Glu Asn Lys Cys Val Gln Ser Asp Ile Asp Ser Phe Phe Lys His Leu
865 870 875 880
Ile Asp Ile Gly Leu Thr Thr Pro Ser Ser Tyr Leu Ser Ile Val Leu
885 890 895
Asp Asp Asn Ile Cys Gly Ala Asp Lys Ala Pro Trp Thr Thr Tyr Thr
900 905 910
Thr Tyr Thr Thr Thr Glu Lys Cys Asn Lys Glu Thr Asp Lys Ser Lys
915 920 925
Leu Gln Gln Cys Asn Thr Ala Val Val Val Asn Val Pro Ser Pro Leu
930 935 940
Gly Asn Thr Pro His Gly Tyr Lys Tyr Ala Cys Gln Cys Lys Ile Pro
945 950 955 960
Thr Asn Glu Glu Thr Cys Asp Asp Arg Lys Glu Tyr Met Asn Gln Trp
965 970 975
Ser Cys Gly Ser Ala Arg Thr Met Lys Arg Gly Tyr Lys Asn Asp Asn
980 985 990
Tyr Glu Leu Cys Lys Tyr Asn Gly Val Asp Val Lys Pro Thr Thr Val
995 1000 1005
Arg Ser Asn Ser Ser Lys Leu Asp Asp Lys Asp Val Thr Phe Phe
1010 1015 1020
Asn Leu Phe Glu Gln Trp Asn Lys Glu Ile Gln Tyr Gln Ile Glu
1025 1030 1035
Gln Tyr Met Thr Asn Thr Lys Ile Ser Cys Asn Asn Glu Lys Asn
1040 1045 1050
Val Leu Ser Arg Val Ser Asp Glu Ala Ala Gln Pro Lys Phe Ser
1055 1060 1065
Asp Asn Glu Arg Asp Arg Asn Ser Ile Thr His Glu Asp Lys Asn
1070 1075 1080
Cys Lys Glu Lys Cys Lys Cys Tyr Ser Leu Trp Ile Glu Lys Ile
1085 1090 1095
Asn Asp Gln Trp Asp Lys Gln Lys Asp Asn Tyr Asn Lys Phe Gln
1100 1105 1110
Arg Lys Gln Ile Tyr Asp Ala Asn Lys Gly Ser Gln Asn Lys Lys
1115 1120 1125
Val Val Ser Leu Ser Asn Phe Leu Phe Phe Ser Cys Trp Glu Glu
1130 1135 1140
Tyr Ile Gln Lys Tyr Phe Asn Gly Asp Trp Ser Lys Ile Lys Asn
1145 1150 1155
Ile Gly Ser Asp Thr Phe Glu Phe Leu Ile Lys Lys Cys Gly Asn
1160 1165 1170
Asp Ser Gly Asp Gly Glu Thr Ile Phe Ser Glu Lys Leu Asn Asn
1175 1180 1185
Ala Glu Lys Lys Cys Lys Glu Asn Glu Ser Thr Asn Asn Lys Met
1190 1195 1200
Lys Ser Ser Glu Thr Ser Cys Asp Cys Ser Glu Pro Ile Tyr Ile
1205 1210 1215
Arg Gly Cys Gln Pro Lys Ile Tyr Asp Gly Lys Ile Phe Pro Gly
1220 1225 1230
Lys Gly Gly Glu Lys Gln Trp Ile Cys Lys Asp Thr Ile Ile His
1235 1240 1245
Gly Asp Thr Asn Gly Ala Cys Ile Pro Pro Arg Thr Gln Asn Leu
1250 1255 1260
Cys Val Gly Glu Leu Trp Asp Lys Arg Tyr Gly Gly Arg Ser Asn
1265 1270 1275
Ile Lys Asn Asp Thr Lys Glu Ser Leu Lys Gln Lys Ile Lys Asn
1280 1285 1290
Ala Ile Gln Lys Glu Thr Glu Leu Leu Tyr Glu Tyr His Asp Lys
1295 1300 1305
Gly Thr Ala Ile Ile Ser Arg Asn Pro Met Lys Gly Gln Lys Glu
1310 1315 1320
Lys Glu Glu Lys Asn Asn Asp Ser Asn Gly Leu Pro Lys Gly Phe
1325 1330 1335
Cys His Ala Val Gln Arg Ser Phe Ile Asp Tyr Lys Asn Met Ile
1340 1345 1350
Leu Gly Thr Ser Val Asn Ile Tyr Glu Tyr Ile Gly Lys Leu Gln
1355 1360 1365
Glu Asp Ile Lys Lys Ile Ile Glu Lys Gly Thr Thr Lys Gln Asn
1370 1375 1380
Gly Lys Thr Val Gly Ser Gly Ala Glu Asn Val Asn Ala Trp Trp
1385 1390 1395
Lys Gly Ile Glu Gly Glu Met Trp Asp Ala Val Arg Cys Ala Ile
1400 1405 1410
Thr Lys Ile Asn Lys Lys Gln Lys Lys Asn Gly Thr Phe Ser Ile
1415 1420 1425
Asp Glu Cys Gly Ile Phe Pro Pro Thr Gly Asn Asp Glu Asp Gln
1430 1435 1440
Ser Val Ser Trp Phe Lys Glu Trp Ser Glu Gln Phe Cys Ile Glu
1445 1450 1455
Arg Leu Gln Tyr Glu Lys Asn Ile Arg Asp Ala Cys Thr Asn Asn
1460 1465 1470
Gly Gln Gly Asp Lys Ile Gln Gly Asp Cys Lys Arg Lys Cys Glu
1475 1480 1485
Glu Tyr Lys Lys Tyr Ile Ser Glu Lys Lys Gln Glu Trp Asp Lys
1490 1495 1500
Gln Lys Thr Lys Tyr Glu Asn Lys Tyr Val Gly Lys Ser Ala Ser
1505 1510 1515
Asp Leu Leu Lys Glu Asn Tyr Pro Glu Cys Ile Ser Ala Asn Phe
1520 1525 1530
Asp Phe Ile Phe Asn Asp Asn Ile Glu Tyr Lys Thr Tyr Tyr Pro
1535 1540 1545
Tyr Gly Asp Tyr Ser Ser Ile Cys Ser Cys Glu Gln Val Lys Tyr
1550 1555 1560
Tyr Glu Tyr Asn Asn Ala Glu Lys Lys Asn Asn Lys Ser Leu Cys
1565 1570 1575
His Glu Lys Gly Asn Asp Arg Thr Trp Ser Lys Lys Tyr Ile Lys
1580 1585 1590
Lys Leu Glu Asn Gly Arg Thr Leu Glu Gly Val Tyr Val Pro Pro
1595 1600 1605
Arg Arg Gln Gln Leu Cys Leu Tyr Glu Leu Phe Pro Ile Ile Ile
1610 1615 1620
Lys Asn Lys Asn Asp Ile Thr Asn Ala Lys Lys Glu Leu Leu Glu
1625 1630 1635
Thr Leu Gln Ile Val Ala Glu Arg Glu Ala Tyr Tyr Leu Trp Lys
1640 1645 1650
Gln Tyr His Ala His Asn Asp Thr Thr Tyr Leu Ala His Lys Lys
1655 1660 1665
Ala Cys Cys Ala Ile Arg Gly Ser Phe Tyr Asp Leu Glu Asp Ile
1670 1675 1680
Ile Lys Gly Asn Asp Leu Val His Asp Glu Tyr Thr Lys Tyr Ile
1685 1690 1695
Asp Ser Lys Leu Asn Glu Ile Phe Asp Ser Ser Asn Lys Asn Asp
1700 1705 1710
Ile Glu Thr Lys Arg Ala Arg Thr Asp Trp Trp Glu Asn Glu Ala
1715 1720 1725
Ile Ala Val Pro Asn Ile Thr Gly Ala Asn Lys Ser Asp Pro Lys
1730 1735 1740
Thr Ile Arg Gln Leu Val Trp Asp Ala Met Gln Ser Gly Val Arg
1745 1750 1755
Lys Ala Ile Asp Glu Glu Lys Glu Lys Lys Lys Pro Asn Glu Asn
1760 1765 1770
Phe Pro Pro Cys Met Gly Val Gln His Ile Gly Ile Ala Lys Pro
1775 1780 1785
Gln Phe Ile Arg Trp Leu Glu Glu Trp Thr Asn Glu Phe Cys Glu
1790 1795 1800
Lys Tyr Thr Lys Tyr Phe Glu Asp Met Lys Ser Asn Cys Asn Leu
1805 1810 1815
Arg Lys Gly Ala Asp Asp Cys Asp Asp Asn Ser Asn Ile Glu Cys
1820 1825 1830
Lys Lys Ala Cys Ala Asn Tyr Thr Asn Trp Leu Asn Pro Lys Arg
1835 1840 1845
Ile Glu Trp Asn Gly Met Ser Asn Tyr Tyr Asn Lys Ile Tyr Arg
1850 1855 1860
Lys Ser Asn Lys Glu Ser Glu Asp Gly Lys Asp Tyr Ser Met Ile
1865 1870 1875
Met Glu Pro Thr Val Ile Asp Tyr Leu Asn Lys Arg Cys Asn Gly
1880 1885 1890
Glu Ile Asn Gly Asn Tyr Ile Cys Cys Ser Cys Lys Asn Ile Gly
1895 1900 1905
Glu Asn Ser Thr Ser Gly Thr Val Asn Lys Lys Leu Gln Lys Lys
1910 1915 1920
Glu Thr Gln Cys Glu Asp Asn Lys Gly Pro Leu Asp Leu Met Asn
1925 1930 1935
Lys Val Leu Asn Lys Met Asp Pro Lys Tyr Ser Glu His Lys Met
1940 1945 1950
Lys Cys Thr Glu Val Tyr Leu Glu His Val Glu Glu Gln Leu Lys
1955 1960 1965
Glu Ile Asp Asn Ala Ile Lys Asp Tyr Lys Leu Tyr Pro Leu Asp
1970 1975 1980
Arg Cys Phe Asp Asp Lys Ser Lys Met Lys Val Cys Asp Leu Ile
1985 1990 1995
Gly Asp Ala Ile Gly Cys Lys His Lys Thr Lys Leu Asp Glu Leu
2000 2005 2010
Asp Glu Trp Asn Asp Val Asp Met Arg Asp Pro Tyr Asn Lys Tyr
2015 2020 2025
Lys Gly Val Leu Ile Pro Pro Arg Arg Arg Gln Leu Cys Phe Ser
2030 2035 2040
Arg Ile Val Arg Gly Pro Ala Asn Leu Arg Asn Leu Lys Glu Phe
2045 2050 2055
Lys Glu Glu Ile Leu Lys Gly Ala Gln Ser Glu Gly Lys Phe Leu
2060 2065 2070
Gly Asn Tyr Tyr Asn Glu Asp Lys Asp Lys Glu Lys Ala Leu Glu
2075 2080 2085
Ala Met Lys Asn Ser Phe Tyr Asp Tyr Glu Tyr Ile Ile Lys Gly
2090 2095 2100
Ser Asp Met Leu Thr Asn Ile Gln Phe Lys Asp Ile Lys Arg Lys
2105 2110 2115
Leu Asp Arg Leu Leu Glu Lys Glu Thr Asn Asn Thr Glu Lys Val
2120 2125 2130
Asp Asp Trp Trp Glu Thr Asn Lys Lys Ser Ile Trp Asn Ala Met
2135 2140 2145
Leu Cys Gly Tyr Lys Lys Ser Gly Asn Lys Ile Ile Asp Pro Ser
2150 2155 2160
Trp Cys Thr Ile Pro Thr Thr Glu Thr Pro Pro Gln Phe Leu Arg
2165 2170 2175
Trp Ile Lys Glu Trp Gly Thr Asn Val Cys Ile Gln Lys Glu Glu
2180 2185 2190
His Lys Glu Tyr Val Lys Ser Lys Cys Ser Asn Val Thr Asn Leu
2195 2200 2205
Gly Ala Gln Glu Ser Glu Ser Lys Asn Cys Thr Ser Glu Ile Lys
2210 2215 2220
Lys Tyr Gln Glu Trp Ser Arg Lys Arg Ser Ile Gln Trp Glu Ala
2225 2230 2235
Ile Ser Glu Gly Tyr Lys Lys Tyr Lys Gly Met Asp Glu Phe Lys
2240 2245 2250
Asn Thr Phe Lys Asn Ile Lys Glu Pro Asp Ala Asn Glu Pro Asn
2255 2260 2265
Ala Asn Glu Tyr Leu Lys Lys His Cys Ser Lys Cys Pro Cys Gly
2270 2275 2280
Phe Asn Asp Met Gln Glu Ile Thr Lys Tyr Thr Asn Ile Gly Asn
2285 2290 2295
Glu Ala Phe Lys Gln Ile Lys Glu Gln Val Asp Ile Pro Ala Glu
2300 2305 2310
Leu Glu Asp Val Ile Tyr Arg Leu Lys His His Glu Tyr Asp Lys
2315 2320 2325
Gly Asn Asp Tyr Ile Cys Asn Lys Tyr Lys Asn Ile Asn Val Asn
2330 2335 2340
Met Lys Lys Asn Asn Asp Asp Thr Trp Thr Asp Leu Val Lys Asn
2345 2350 2355
Ser Ser Asp Ile Asn Lys Gly Val Leu Leu Pro Pro Arg Arg Lys
2360 2365 2370
Asn Leu Phe Leu Lys Ile Asp Glu Ser Asp Ile Cys Lys Tyr Lys
2375 2380 2385
Arg Asp Pro Lys Leu Phe Lys Asp Phe Ile Tyr Ser Ser Ala Ile
2390 2395 2400
Ser Glu Val Glu Arg Leu Lys Lys Val Tyr Gly Glu Ala Lys Thr
2405 2410 2415
Lys Val Val His Ala Met Lys Tyr Ser Phe Ala Asp Ile Gly Ser
2420 2425 2430
Ile Ile Lys Gly Asp Asp Met Met Glu Asn Asn Ser Ser Asp Lys
2435 2440 2445
Ile Gly Lys Ile Leu Gly Asp Gly Val Gly Gln Asn Glu Lys Arg
2450 2455 2460
Lys Lys Trp Trp Asp Met Asn Lys Tyr His Ile Trp Glu Ser Met
2465 2470 2475
Leu Cys Gly Tyr Lys His Ala Tyr Gly Asn Ile Ser Glu Asn Asp
2480 2485 2490
Arg Lys Met Leu Asp Ile Pro Asn Asn Asp Asp Glu His Gln Phe
2495 2500 2505
Leu Arg Trp Phe Gln Glu Trp Thr Glu Asn Phe Cys Thr Lys Arg
2510 2515 2520
Asn Glu Leu Tyr Glu Asn Met Val Thr Ala Cys Asn Ser Ala Lys
2525 2530 2535
Cys Asn Thr Ser Asn Gly Ser Val Asp Lys Lys Glu Cys Thr Glu
2540 2545 2550
Ala Cys Lys Asn Tyr Ser Asn Phe Ile Leu Ile Lys Lys Lys Glu
2555 2560 2565
Tyr Gln Ser Leu Asn Ser Gln Tyr Asp Met Asn Tyr Lys Glu Thr
2570 2575 2580
Lys Ala Glu Lys Lys Glu Ser Pro Glu Tyr Phe Lys Asp Lys Cys
2585 2590 2595
Asn Gly Glu Cys Ser Cys Leu Ser Glu Tyr Phe Lys Asp Glu Thr
2600 2605 2610
Arg Trp Lys Asn Pro Tyr Glu Thr Leu Asp Asp Thr Glu Val Lys
2615 2620 2625
Asn Asn Cys Met Cys Lys Pro Pro Pro Pro Ala Ser Asn Asn Thr
2630 2635 2640
Ser Asp Ile Leu Gln Lys Thr Ile Pro Phe Gly Ile Ala Leu Ala
2645 2650 2655
Leu Gly Ser Ile Ala Phe Leu Phe Met Lys Lys Lys Pro Lys Thr
2660 2665 2670
Pro Val Asp Leu Leu Arg Val Leu Asp Ile Pro Lys Gly Asp Tyr
2675 2680 2685
Gly Ile Pro Thr Pro Lys Ser Ser Asn Arg Tyr Ile Pro Tyr Ala
2690 2695 2700
Ser Asp Arg Tyr Lys Gly Lys Thr Tyr Ile Tyr Met Glu Gly Asp
2705 2710 2715
Thr Ser Gly Asp Asp Asp Lys Tyr Ile Trp Asp Leu
2720 2725 2730
<210> 56
<211> 2734
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 56
Met Asp Ser Thr Ser Thr Ile Ala Asn Lys Ile Glu Glu Tyr Leu Gly
1 5 10 15
Ala Lys Ser Asp Asp Ser Lys Ile Asp Glu Leu Leu Lys Ala Asp Pro
20 25 30
Ser Glu Val Glu Tyr Tyr Arg Ser Gly Gly Asp Gly Asp Tyr Leu Lys
35 40 45
Asn Asn Ile Cys Lys Ile Thr Val Asn His Ser Asp Ser Gly Lys Tyr
50 55 60
Asp Pro Cys Glu Lys Lys Leu Pro Pro Tyr Asp Asp Asn Asp Gln Trp
65 70 75 80
Lys Cys Gln Gln Asn Ser Ser Asp Gly Ser Gly Lys Pro Glu Asn Ile
85 90 95
Cys Val Pro Pro Arg Arg Glu Arg Leu Cys Thr Tyr Asn Leu Glu Asn
100 105 110
Leu Lys Phe Asp Lys Ile Arg Asp Asn Asn Ala Phe Leu Ala Asp Val
115 120 125
Leu Leu Thr Ala Arg Asn Glu Gly Glu Lys Ile Val Gln Asn His Pro
130 135 140
Asp Thr Asn Ser Ser Asn Val Cys Asn Ala Leu Glu Arg Ser Phe Ala
145 150 155 160
Asp Leu Ala Asp Ile Ile Arg Gly Thr Asp Gln Trp Lys Gly Thr Asn
165 170 175
Ser Asn Leu Glu Lys Asn Leu Lys Gln Met Phe Ala Lys Ile Arg Glu
180 185 190
Asn Asp Lys Val Leu Gln Asp Lys Tyr Pro Lys Asp Gln Lys Tyr Thr
195 200 205
Lys Leu Arg Glu Ala Trp Trp Asn Ala Asn Arg Gln Lys Val Trp Glu
210 215 220
Val Ile Thr Cys Gly Ala Arg Ser Asn Asp Leu Leu Ile Lys Arg Gly
225 230 235 240
Trp Arg Thr Ser Gly Lys Ser Asp Arg Lys Lys Asn Phe Glu Leu Cys
245 250 255
Arg Lys Cys Gly His Tyr Glu Lys Glu Val Pro Thr Lys Leu Asp Tyr
260 265 270
Val Pro Gln Phe Leu Arg Trp Leu Thr Glu Trp Ile Glu Asp Phe Tyr
275 280 285
Arg Glu Lys Gln Asn Leu Ile Asp Asp Met Glu Arg His Arg Glu Glu
290 295 300
Cys Thr Arg Glu Asp His Lys Ser Lys Glu Gly Thr Ser Tyr Cys Ser
305 310 315 320
Thr Cys Lys Asp Lys Cys Lys Lys Tyr Cys Glu Cys Val Lys Lys Trp
325 330 335
Lys Thr Glu Trp Glu Asn Gln Glu Asn Lys Tyr Lys Asp Leu Tyr Glu
340 345 350
Gln Asn Lys Asn Lys Thr Ser Gln Lys Asn Thr Ser Arg Tyr Asp Asp
355 360 365
Tyr Val Lys Asp Phe Phe Glu Lys Leu Glu Ala Asn Tyr Ser Ser Leu
370 375 380
Glu Asn Tyr Ile Lys Gly Asp Pro Tyr Phe Ala Glu Tyr Ala Thr Lys
385 390 395 400
Leu Ser Phe Ile Leu Asn Pro Ser Asp Ala Asn Asn Pro Ser Gly Glu
405 410 415
Thr Ala Asn His Asn Asp Glu Ala Cys Asn Cys Asn Glu Ser Gly Ile
420 425 430
Ser Ser Val Gly Gln Ala Gln Thr Ser Gly Pro Ser Ser Asn Lys Thr
435 440 445
Cys Ile Thr His Ser Ser Ile Lys Thr Asn Lys Lys Lys Glu Cys Lys
450 455 460
Asp Val Lys Leu Gly Val Arg Glu Asn Asp Lys Asp Leu Lys Ile Cys
465 470 475 480
Val Ile Glu Asp Thr Ser Leu Ser Gly Val Asp Asn Cys Cys Cys Gln
485 490 495
Asp Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Arg Gly
500 505 510
Ser Ser Ser Asn Asp Ser Cys Asp Asn Lys Asn Gln Asp Glu Cys Gln
515 520 525
Lys Lys Leu Glu Lys Val Phe Ala Ser Leu Thr Asn Gly Tyr Lys Cys
530 535 540
Asp Lys Cys Lys Ser Gly Thr Ser Arg Ser Lys Lys Lys Trp Ile Trp
545 550 555 560
Lys Lys Ser Ser Gly Asn Glu Glu Gly Leu Gln Glu Glu Tyr Ala Asn
565 570 575
Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu Gly Asn Leu
580 585 590
Pro Lys Leu Glu Asn Val Cys Glu Asp Val Lys Asp Ile Asn Phe Asp
595 600 605
Thr Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Val Ser Phe His Glu
610 615 620
Gly Lys Asn Leu Lys Lys Arg Tyr Pro Gln Asn Lys Asn Ser Gly Asn
625 630 635 640
Lys Glu Asn Leu Cys Lys Ala Leu Glu Tyr Ser Phe Ala Asp Tyr Gly
645 650 655
Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp
660 665 670
Leu Glu Leu Asn Leu Gln Asn Asn Phe Gly Lys Leu Phe Gly Lys Tyr
675 680 685
Ile Lys Lys Asn Asn Thr Ala Glu Gln Asp Thr Ser Tyr Ser Ser Leu
690 695 700
Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp
705 710 715 720
Thr Ala Met Lys His Gly Ala Glu Met Asn Ile Thr Thr Cys Asn Ala
725 730 735
Asp Gly Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp Ile Pro Thr
740 745 750
Ile Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu
755 760 765
Asn Phe Cys Glu Gln Arg Gln Ala Lys Val Lys Asp Val Ile Thr Asn
770 775 780
Cys Lys Ser Cys Lys Glu Ser Gly Asn Lys Cys Lys Thr Glu Cys Lys
785 790 795 800
Thr Lys Cys Lys Asp Glu Cys Glu Lys Tyr Lys Lys Phe Ile Glu Ala
805 810 815
Cys Gly Thr Ala Gly Gly Gly Ile Gly Thr Ala Gly Ser Pro Trp Ser
820 825 830
Lys Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys His Ile Glu Asp
835 840 845
Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Thr Ser Ser
850 855 860
Thr Thr Asn Ala Ala Ala Ser Thr Asp Glu Asn Lys Cys Val Gln Ser
865 870 875 880
Asp Ile Asp Ser Phe Phe Lys His Leu Ile Asp Ile Gly Leu Thr Thr
885 890 895
Pro Ser Ser Tyr Leu Ser Asn Val Leu Asp Asp Asn Ile Cys Gly Ala
900 905 910
Asp Lys Ala Pro Trp Thr Thr Tyr Thr Thr Tyr Thr Thr Thr Glu Lys
915 920 925
Cys Asn Lys Glu Arg Asp Lys Ser Lys Ser Gln Ser Ser Asp Thr Leu
930 935 940
Val Val Val Asn Val Pro Ser Pro Leu Gly Asn Thr Pro Tyr Arg Tyr
945 950 955 960
Lys Tyr Ala Cys Gln Cys Lys Ile Pro Thr Asn Glu Glu Thr Cys Asp
965 970 975
Asp Arg Lys Glu Tyr Met Asn Gln Trp Ser Cys Gly Ser Ala Arg Thr
980 985 990
Met Lys Arg Gly Tyr Lys Asn Asp Asn Tyr Glu Leu Cys Lys Tyr Asn
995 1000 1005
Gly Val Asp Val Lys Pro Thr Thr Val Arg Ser Asn Ser Ser Lys
1010 1015 1020
Leu Asp Gly Asn Asp Val Thr Phe Phe Asn Leu Phe Glu Gln Trp
1025 1030 1035
Asn Lys Glu Ile Gln Tyr Gln Ile Glu Gln Tyr Met Thr Asn Ala
1040 1045 1050
Asn Ile Ser Cys Ile Asp Glu Lys Glu Val Leu Asp Ser Val Ser
1055 1060 1065
Asp Glu Gly Thr Pro Lys Val Arg Gly Gly Tyr Glu Asp Gly Arg
1070 1075 1080
Asn Asn Asn Thr Asp Gln Gly Thr Asn Cys Lys Glu Lys Cys Lys
1085 1090 1095
Cys Tyr Lys Leu Trp Ile Glu Lys Ile Asn Asp Gln Trp Gly Lys
1100 1105 1110
Gln Lys Asp Asn Tyr Asn Lys Phe Arg Ser Lys Gln Ile Tyr Asp
1115 1120 1125
Ala Asn Lys Gly Ser Gln Asn Lys Lys Val Val Ser Leu Ser Asn
1130 1135 1140
Phe Leu Phe Phe Ser Cys Trp Glu Glu Tyr Ile Gln Lys Tyr Phe
1145 1150 1155
Asn Gly Asp Trp Ser Lys Ile Lys Asn Ile Gly Ser Asp Thr Phe
1160 1165 1170
Glu Phe Leu Ile Lys Lys Cys Gly Asn Asn Ser Ala His Gly Glu
1175 1180 1185
Glu Ile Phe Asn Glu Lys Leu Lys Asn Ala Glu Lys Lys Cys Lys
1190 1195 1200
Glu Asn Glu Ser Thr Asp Thr Asn Ile Asn Lys Ser Glu Thr Ser
1205 1210 1215
Cys Asp Leu Asn Ala Thr Asn Tyr Ile Arg Gly Cys Gln Ser Lys
1220 1225 1230
Thr Tyr Asp Gly Lys Ile Phe Pro Gly Lys Gly Gly Glu Lys Gln
1235 1240 1245
Trp Ile Cys Lys Asp Thr Ile Ile His Gly Asp Thr Asn Gly Ala
1250 1255 1260
Cys Ile Pro Pro Arg Thr Gln Asn Leu Cys Val Gly Glu Leu Trp
1265 1270 1275
Asp Lys Ser Tyr Gly Gly Arg Ser Asn Ile Lys Asn Asp Thr Lys
1280 1285 1290
Glu Leu Leu Lys Glu Lys Ile Lys Asn Ala Ile His Lys Glu Thr
1295 1300 1305
Glu Leu Leu Tyr Glu Tyr His Asp Thr Gly Thr Ala Ile Ile Ser
1310 1315 1320
Lys Asn Asp Lys Lys Gly Gln Lys Gly Lys Asn Asp Pro Asn Gly
1325 1330 1335
Leu Pro Lys Gly Phe Cys His Ala Val Gln Arg Ser Phe Ile Asp
1340 1345 1350
Tyr Lys Asn Met Ile Leu Gly Thr Ser Val Asn Ile Tyr Glu His
1355 1360 1365
Ile Gly Lys Leu Gln Glu Asp Ile Lys Lys Ile Ile Glu Lys Gly
1370 1375 1380
Thr Pro Gln Gln Lys Asp Lys Ile Gly Gly Val Gly Ser Ser Thr
1385 1390 1395
Glu Asn Val Asn Ala Trp Trp Lys Gly Ile Glu Arg Glu Met Trp
1400 1405 1410
Asp Ala Val Arg Cys Ala Ile Thr Lys Ile Asn Lys Lys Asn Asn
1415 1420 1425
Asn Ser Ile Phe Asn Gly Asp Glu Cys Gly Val Ser Pro Pro Thr
1430 1435 1440
Gly Asn Asp Glu Asp Gln Ser Val Ser Trp Phe Lys Glu Trp Gly
1445 1450 1455
Glu Gln Phe Cys Ile Glu Arg Leu Arg Tyr Glu Gln Asn Ile Arg
1460 1465 1470
Glu Ala Cys Thr Ile Asn Gly Lys Asn Glu Lys Lys Cys Ile Asn
1475 1480 1485
Ser Lys Ser Gly Gln Gly Asp Lys Ile Gln Gly Ala Cys Lys Arg
1490 1495 1500
Lys Cys Glu Lys Tyr Lys Lys Tyr Ile Ser Glu Lys Lys Gln Glu
1505 1510 1515
Trp Asp Lys Gln Lys Thr Lys Tyr Glu Asn Lys Tyr Val Gly Lys
1520 1525 1530
Ser Ala Ser Asp Leu Leu Lys Glu Asn Tyr Pro Glu Cys Ile Ser
1535 1540 1545
Ala Asn Phe Asp Phe Ile Phe Asn Asp Asn Ile Glu Tyr Lys Thr
1550 1555 1560
Tyr Tyr Pro Tyr Gly Asp Tyr Ser Ser Ile Cys Ser Cys Glu Gln
1565 1570 1575
Val Lys Tyr Tyr Lys Tyr Asn Asn Ala Glu Lys Lys Asn Asn Lys
1580 1585 1590
Ser Leu Cys Tyr Glu Lys Asp Asn Asp Met Thr Trp Ser Lys Lys
1595 1600 1605
Tyr Ile Lys Lys Leu Glu Asn Gly Arg Ser Leu Glu Gly Val Tyr
1610 1615 1620
Val Pro Pro Arg Arg Gln Gln Leu Cys Leu Tyr Glu Leu Phe Pro
1625 1630 1635
Ile Ile Ile Lys Asn Glu Glu Gly Met Glu Lys Ala Lys Glu Glu
1640 1645 1650
Leu Leu Glu Thr Leu Gln Ile Val Ala Glu Arg Glu Ala Tyr Tyr
1655 1660 1665
Leu Trp Lys Gln Tyr Asn Pro Thr Gly Lys Gly Ile Asp Asp Ala
1670 1675 1680
Asn Lys Lys Ala Cys Cys Ala Ile Arg Gly Ser Phe Tyr Asp Leu
1685 1690 1695
Glu Asp Ile Ile Lys Gly Asn Asp Leu Val His Asp Glu Tyr Thr
1700 1705 1710
Lys Tyr Ile Asp Ser Lys Leu Asn Glu Ile Phe Gly Ser Ser Asp
1715 1720 1725
Thr Asn Asp Ile Asp Thr Lys Arg Ala Arg Thr Asp Trp Trp Glu
1730 1735 1740
Asn Glu Thr Ile Thr Asn Gly Thr Asp Arg Lys Thr Ile Arg Gln
1745 1750 1755
Leu Val Trp Asp Ala Met Gln Ser Gly Val Arg Tyr Ala Val Glu
1760 1765 1770
Glu Lys Asn Glu Asn Phe Pro Leu Cys Met Gly Val Glu His Ile
1775 1780 1785
Gly Ile Ala Lys Pro Gln Phe Ile Arg Trp Leu Glu Glu Trp Thr
1790 1795 1800
Asn Glu Phe Cys Glu Lys Tyr Thr Lys Tyr Phe Glu Asp Met Lys
1805 1810 1815
Ser Lys Cys Asp Pro Pro Lys Arg Ala Asp Thr Cys Gly Asp Asn
1820 1825 1830
Ser Asn Ile Glu Cys Lys Lys Ala Cys Ala Asn Tyr Thr Asn Trp
1835 1840 1845
Leu Asn Pro Lys Arg Ile Glu Trp Asn Gly Met Ser Asn Tyr Tyr
1850 1855 1860
Asn Lys Ile Tyr Arg Lys Ser Asn Lys Glu Ser Glu Gly Gly Lys
1865 1870 1875
Asp Tyr Ser Met Ile Met Ala Pro Thr Val Ile Asp Tyr Leu Asn
1880 1885 1890
Lys Arg Cys His Gly Glu Ile Asn Gly Asn Tyr Ile Cys Cys Ser
1895 1900 1905
Cys Lys Asn Ile Gly Ala Tyr Asn Thr Thr Ser Gly Thr Val Asn
1910 1915 1920
Lys Lys Leu Gln Lys Lys Glu Thr Glu Cys Glu Glu Glu Lys Gly
1925 1930 1935
Pro Leu Asp Leu Met Asn Glu Val Leu Asn Lys Met Asp Lys Lys
1940 1945 1950
Tyr Ser Ala His Lys Met Lys Cys Thr Glu Val Tyr Leu Glu His
1955 1960 1965
Val Glu Glu Gln Leu Asn Glu Ile Asp Asn Ala Ile Lys Asp Tyr
1970 1975 1980
Lys Leu Tyr Pro Leu Asp Arg Cys Phe Asp Asp Gln Thr Lys Met
1985 1990 1995
Lys Val Cys Asp Leu Ile Ala Asp Ala Ile Gly Cys Lys Asp Lys
2000 2005 2010
Thr Lys Leu Asp Glu Leu Asp Glu Trp Asn Asp Met Asp Leu Arg
2015 2020 2025
Gly Thr Tyr Asn Lys His Lys Gly Val Leu Ile Pro Pro Arg Arg
2030 2035 2040
Arg Gln Leu Cys Phe Ser Arg Ile Val Arg Gly Pro Ala Asn Leu
2045 2050 2055
Arg Ser Leu Asn Glu Phe Lys Glu Glu Ile Leu Lys Gly Ala Gln
2060 2065 2070
Ser Glu Gly Lys Phe Leu Gly Asn Tyr Tyr Lys Glu His Lys Asp
2075 2080 2085
Lys Glu Lys Ala Leu Glu Ala Met Lys Asn Ser Phe Tyr Asp Tyr
2090 2095 2100
Glu Asp Ile Ile Lys Gly Thr Asp Met Leu Thr Asn Ile Glu Phe
2105 2110 2115
Lys Asp Ile Lys Ile Lys Leu Asp Arg Leu Leu Glu Lys Glu Thr
2120 2125 2130
Asn Asn Thr Lys Lys Ala Glu Asp Trp Trp Lys Thr Asn Lys Lys
2135 2140 2145
Ser Ile Trp Asn Ala Met Leu Cys Gly Tyr Lys Lys Ser Gly Asn
2150 2155 2160
Lys Ile Ile Asp Pro Ser Trp Cys Thr Ile Pro Thr Thr Glu Thr
2165 2170 2175
Pro Pro Gln Phe Leu Arg Trp Ile Lys Glu Trp Gly Thr Asn Val
2180 2185 2190
Cys Ile Gln Lys Gln Glu His Lys Glu Tyr Val Lys Ser Lys Cys
2195 2200 2205
Ser Asn Val Thr Asn Leu Gly Ala Gln Ala Ser Glu Ser Asn Asn
2210 2215 2220
Cys Thr Ser Glu Ile Lys Lys Tyr Gln Glu Trp Ser Arg Lys Arg
2225 2230 2235
Ser Ile Arg Trp Glu Thr Ile Ser Lys Arg Tyr Lys Lys Tyr Lys
2240 2245 2250
Arg Met Asp Ile Leu Lys Asp Val Lys Glu Pro Asp Ala Asn Thr
2255 2260 2265
Tyr Leu Arg Glu His Cys Ser Lys Cys Pro Cys Gly Phe Asn Asp
2270 2275 2280
Met Glu Glu Met Asn Asn Asn Glu Asp Asn Glu Lys Glu Ala Phe
2285 2290 2295
Lys Gln Ile Lys Glu Gln Val Lys Ile Pro Ala Glu Leu Glu Asp
2300 2305 2310
Val Ile Tyr Arg Ile Lys His His Glu Tyr Asp Lys Gly Asn Asp
2315 2320 2325
Tyr Ile Cys Asn Lys Tyr Lys Asn Ile His Asp Arg Met Lys Lys
2330 2335 2340
Asn Asn Gly Asn Phe Val Thr Asp Asn Phe Val Lys Lys Ser Trp
2345 2350 2355
Glu Ile Ser Asn Gly Val Leu Ile Pro Pro Arg Arg Lys Asn Leu
2360 2365 2370
Phe Leu Tyr Ile Asp Pro Ser Lys Ile Cys Glu Tyr Lys Lys Asp
2375 2380 2385
Pro Lys Leu Phe Lys Asp Phe Ile Tyr Trp Ser Ala Phe Thr Glu
2390 2395 2400
Val Glu Arg Leu Lys Lys Ala Tyr Gly Gly Ala Arg Ala Lys Val
2405 2410 2415
Val His Ala Met Lys Tyr Ser Phe Thr Asp Ile Gly Ser Ile Ile
2420 2425 2430
Lys Gly Asp Asp Met Met Glu Lys Asn Ser Ser Asp Lys Ile Gly
2435 2440 2445
Lys Ile Leu Gly Asp Thr Asp Gly Gln Asn Glu Lys Arg Lys Lys
2450 2455 2460
Trp Trp Asp Met Asn Lys Tyr His Ile Trp Glu Ser Met Leu Cys
2465 2470 2475
Gly Tyr Arg Glu Ala Glu Gly Asp Thr Glu Thr Asn Glu Asn Cys
2480 2485 2490
Arg Phe Pro Asp Ile Glu Ser Val Pro Gln Phe Leu Arg Trp Phe
2495 2500 2505
Gln Glu Trp Ser Glu Asn Phe Cys Asp Arg Arg Gln Lys Leu Tyr
2510 2515 2520
Asp Lys Leu Asn Ser Glu Cys Ile Ser Ala Glu Cys Thr Asn Gly
2525 2530 2535
Ser Val Asp Asn Ser Lys Cys Thr His Ala Cys Val Asn Tyr Lys
2540 2545 2550
Asn Tyr Ile Leu Thr Lys Lys Thr Glu Tyr Glu Ile Gln Thr Asn
2555 2560 2565
Lys Tyr Asp Asn Glu Phe Lys Asn Lys Asn Ser Asn Asp Lys Asp
2570 2575 2580
Ala Pro Asp Tyr Leu Lys Glu Lys Cys Asn Asp Asn Lys Cys Glu
2585 2590 2595
Cys Leu Asn Lys His Ile Asp Asp Lys Asn Lys Thr Trp Lys Asn
2600 2605 2610
Pro Tyr Glu Thr Leu Glu Asp Thr Phe Lys Ser Lys Cys Asp Cys
2615 2620 2625
Pro Lys Pro Leu Pro Ser Pro Ile Lys Pro Asp Asp Leu Pro Pro
2630 2635 2640
Gln Ala Asp Glu Pro Phe Asp Pro Thr Ile Leu Gln Thr Thr Ile
2645 2650 2655
Pro Phe Gly Ile Ala Leu Ala Leu Gly Ser Ile Ala Phe Leu Phe
2660 2665 2670
Met Lys Val Ile Tyr Ile Tyr Ile Tyr Val Cys Cys Ile Cys Met
2675 2680 2685
Tyr Val Cys Met Tyr Val Cys Met Tyr Val Cys Met Tyr Val Cys
2690 2695 2700
Met Tyr Val Cys Met His Val Cys Met Leu Cys Val Tyr Val Ile
2705 2710 2715
Tyr Val Phe Lys Ile Cys Ile Tyr Ile Glu Lys Glu Lys Arg Lys
2720 2725 2730
Lys
<210> 57
<211> 58
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 57
Arg Pro Asp Phe Cys Leu Glu Pro Pro Tyr Thr Gly Pro Cys Lys Ala
1 5 10 15
Arg Ile Ile Arg Tyr Phe Tyr Asn Ala Lys Ala Gly Leu Cys Gln Thr
20 25 30
Phe Val Tyr Gly Gly Cys Arg Ala Lys Arg Asn Asn Phe Lys Ser Ala
35 40 45
Glu Asp Cys Met Arg Thr Cys Gly Gly Ala
50 55
<210> 58
<211> 324
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 58
Arg His Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly Tyr
1 5 10 15
Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser Trp
20 25 30
Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly Ser
35 40 45
Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala Arg
50 55 60
Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val Arg Gln
65 70 75 80
Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Asn Gly Pro Ala Asp Ser
85 90 95
Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu Phe Leu
100 105 110
Gly Asp Gly Gly Asp Ile Ser Phe Ser Thr Arg Gly Thr Gln Asn Trp
115 120 125
Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg Gly
130 135 140
Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln Ser
145 150 155 160
Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala Ile
165 170 175
Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly Tyr
180 185 190
Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly Ala
195 200 205
Leu Leu Arg Val Tyr Val Pro Arg Ser Ser Leu Pro Gly Phe Tyr Arg
210 215 220
Thr Ser Leu Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu Val Glu Arg
225 230 235 240
Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala Ile Thr Gly Pro
245 250 255
Glu Glu Glu Gly Gly Arg Leu Glu Thr Ile Leu Gly Trp Pro Leu Ala
260 265 270
Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr Asp Pro Arg Asn
275 280 285
Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys Glu Gln Ala
290 295 300
Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly Lys Pro Pro Arg
305 310 315 320
Lys Asp Glu Leu
<210> 59
<211> 231
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 59
Arg His Arg Gln Pro Arg Gly Trp Glu Gln Leu Tyr Pro Thr Gly Ala
1 5 10 15
Glu Phe Leu Gly Asp Gly Gly Asp Ile Ser Phe Ser Thr Arg Gly Thr
20 25 30
Gln Asn Trp Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu
35 40 45
Glu Arg Gly Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala
50 55 60
Ala Gln Ser Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu
65 70 75 80
Asp Ala Ile Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala
85 90 95
Tyr Gly Tyr Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg
100 105 110
Asn Gly Ala Leu Leu Arg Val Tyr Val Pro Arg Ser Ser Leu Pro Gly
115 120 125
Phe Tyr Arg Thr Ser Leu Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu
130 135 140
Val Glu Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala Ile
145 150 155 160
Thr Gly Pro Glu Glu Glu Gly Gly Arg Leu Glu Thr Ile Leu Gly Trp
165 170 175
Pro Leu Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr Asp
180 185 190
Pro Arg Asn Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys
195 200 205
Glu Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly Lys
210 215 220
Pro Pro Arg Lys Asp Glu Leu
225 230
<210> 60
<211> 929
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 60
Arg Pro Asp Phe Cys Leu Glu Pro Pro Tyr Thr Gly Pro Cys Lys Ala
1 5 10 15
Arg Ile Ile Arg Tyr Phe Tyr Asn Ala Lys Ala Gly Leu Cys Gln Thr
20 25 30
Phe Val Tyr Gly Gly Cys Arg Ala Lys Arg Asn Asn Phe Lys Ser Ala
35 40 45
Glu Asp Cys Met Arg Thr Cys Gly Gly Ala Asn Tyr Ile Lys Gly Asp
50 55 60
Pro Tyr Phe Ala Glu Tyr Ala Thr Lys Leu Ser Phe Ile Leu Asn Pro
65 70 75 80
Ser Asp Ala Asn Asn Pro Ser Gly Glu Thr Ala Asn His Asn Asp Glu
85 90 95
Ala Cys Asn Cys Asn Glu Ser Gly Ile Ser Ser Val Gly Gln Ala Gln
100 105 110
Thr Ser Gly Pro Ser Ser Asn Lys Thr Cys Ile Thr His Ser Ser Ile
115 120 125
Lys Thr Asn Lys Lys Lys Glu Cys Lys Asp Val Lys Leu Gly Val Arg
130 135 140
Glu Asn Asp Lys Asp Leu Lys Ile Cys Val Ile Glu Asp Thr Ser Leu
145 150 155 160
Ser Gly Val Asp Asn Cys Cys Cys Gln Asp Leu Leu Gly Ile Leu Gln
165 170 175
Glu Asn Cys Ser Asp Asn Lys Arg Gly Ser Ser Ser Asn Asp Ser Cys
180 185 190
Asp Asn Lys Asn Gln Asp Glu Cys Gln Lys Lys Leu Glu Lys Val Phe
195 200 205
Ala Ser Leu Thr Asn Gly Tyr Lys Cys Asp Lys Cys Lys Ser Gly Thr
210 215 220
Ser Arg Ser Lys Lys Lys Trp Ile Trp Lys Lys Ser Ser Gly Asn Glu
225 230 235 240
Glu Gly Leu Gln Glu Glu Tyr Ala Asn Thr Ile Gly Leu Pro Pro Arg
245 250 255
Thr Gln Ser Leu Tyr Leu Gly Asn Leu Pro Lys Leu Glu Asn Val Cys
260 265 270
Glu Asp Val Lys Asp Ile Asn Phe Asp Thr Lys Glu Lys Phe Leu Ala
275 280 285
Gly Cys Leu Ile Val Ser Phe His Glu Gly Lys Asn Leu Lys Lys Arg
290 295 300
Tyr Pro Gln Asn Lys Asn Ser Gly Asn Lys Glu Asn Leu Cys Lys Ala
305 310 315 320
Leu Glu Tyr Ser Phe Ala Asp Tyr Gly Asp Leu Ile Lys Gly Thr Ser
325 330 335
Ile Trp Asp Asn Glu Tyr Thr Lys Asp Leu Glu Leu Asn Leu Gln Asn
340 345 350
Asn Phe Gly Lys Leu Phe Gly Lys Tyr Ile Lys Lys Asn Asn Thr Ala
355 360 365
Glu Gln Asp Thr Ser Tyr Ser Ser Leu Asp Glu Leu Arg Glu Ser Trp
370 375 380
Trp Asn Thr Asn Lys Lys Tyr Ile Trp Thr Ala Met Lys His Gly Ala
385 390 395 400
Glu Met Asn Ile Thr Thr Cys Asn Ala Asp Gly Ser Val Thr Gly Ser
405 410 415
Gly Ser Ser Cys Asp Asp Ile Pro Thr Ile Asp Leu Ile Pro Gln Tyr
420 425 430
Leu Arg Phe Leu Gln Glu Trp Val Glu Asn Phe Cys Glu Gln Arg Gln
435 440 445
Ala Lys Val Lys Asp Val Ile Thr Asn Cys Lys Ser Cys Lys Glu Ser
450 455 460
Gly Asn Lys Cys Lys Thr Glu Cys Lys Thr Lys Cys Lys Asp Glu Cys
465 470 475 480
Glu Lys Tyr Lys Lys Phe Ile Glu Ala Cys Gly Thr Ala Gly Gly Gly
485 490 495
Ile Gly Thr Ala Gly Ser Pro Trp Ser Lys Arg Trp Asp Gln Ile Tyr
500 505 510
Lys Arg Tyr Ser Lys His Ile Glu Asp Ala Lys Arg Asn Arg Lys Ala
515 520 525
Gly Thr Lys Asn Cys Gly Thr Ser Ser Thr Thr Asn Ala Ala Ala Ser
530 535 540
Thr Asp Glu Asn Lys Cys Val Gln Ser Asp Ile Asp Ser Phe Phe Lys
545 550 555 560
His Leu Ile Asp Ile Gly Leu Thr Thr Pro Ser Ser Tyr Leu Ser Asn
565 570 575
Val Leu Asp Asp Asn Ile Cys Gly Ala Asp Lys Ala Pro Trp Thr Thr
580 585 590
Tyr Thr Thr Tyr Thr Thr Thr Glu Lys Cys Asn Lys Glu Arg Asp Lys
595 600 605
Ser Lys Ser Gln Ser Ser Asp Thr Leu Val Val Val Asn Val Pro Ser
610 615 620
Pro Leu Gly Asn Thr Pro Tyr Arg Tyr Lys Tyr Ala Cys Gln Cys Lys
625 630 635 640
Ile Pro Thr Asn Glu Glu Thr Cys Asp Asp Arg Lys Glu Tyr Met Asn
645 650 655
Gln Trp Ser Cys Gly Ser Ala Arg Thr Met Lys Arg Gly Tyr Lys Asn
660 665 670
Asp Asn Tyr Glu Leu Cys Lys Tyr Asn Gly Val Asp Val Lys Pro Thr
675 680 685
Thr Val Arg Ser Asn Ser Ser Lys Leu Asp Arg His Arg Gln Pro Arg
690 695 700
Gly Trp Glu Gln Leu Tyr Pro Thr Gly Ala Glu Phe Leu Gly Asp Gly
705 710 715 720
Gly Asp Ile Ser Phe Ser Thr Arg Gly Thr Gln Asn Trp Thr Val Glu
725 730 735
Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg Gly Tyr Val Phe
740 745 750
Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln Ser Ile Val Phe
755 760 765
Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala Ile Trp Arg Gly
770 775 780
Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly Tyr Ala Gln Asp
785 790 795 800
Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly Ala Leu Leu Arg
805 810 815
Val Tyr Val Pro Arg Ser Ser Leu Pro Gly Phe Tyr Arg Thr Ser Leu
820 825 830
Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu Val Glu Arg Leu Ile Gly
835 840 845
His Pro Leu Pro Leu Arg Leu Asp Ala Ile Thr Gly Pro Glu Glu Glu
850 855 860
Gly Gly Arg Leu Glu Thr Ile Leu Gly Trp Pro Leu Ala Glu Arg Thr
865 870 875 880
Val Val Ile Pro Ser Ala Ile Pro Thr Asp Pro Arg Asn Val Gly Gly
885 890 895
Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys Glu Gln Ala Ile Ser Ala
900 905 910
Leu Pro Asp Tyr Ala Ser Gln Pro Gly Lys Pro Pro Arg Lys Asp Glu
915 920 925
Leu
<210> 61
<211> 1045
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 61
Arg Pro Asp Phe Cys Leu Glu Pro Pro Tyr Thr Gly Pro Cys Lys Ala
1 5 10 15
Arg Ile Ile Arg Tyr Phe Tyr Asn Ala Lys Ala Gly Leu Cys Gln Thr
20 25 30
Phe Val Tyr Gly Gly Cys Arg Ala Lys Arg Asn Asn Phe Lys Ser Ala
35 40 45
Glu Asp Cys Met Arg Thr Cys Gly Gly Ala Asn Tyr Ile Lys Gly Asp
50 55 60
Pro Tyr Phe Ala Glu Tyr Ala Thr Lys Leu Ser Phe Ile Leu Asn Pro
65 70 75 80
Ser Asp Ala Asn Asn Pro Ser Gly Glu Thr Ala Asn His Asn Asp Glu
85 90 95
Ala Cys Asn Cys Asn Glu Ser Gly Ile Ser Ser Val Gly Gln Ala Gln
100 105 110
Thr Ser Gly Pro Ser Ser Asn Lys Thr Cys Ile Thr His Ser Ser Ile
115 120 125
Lys Thr Asn Lys Lys Lys Glu Cys Lys Asp Val Lys Leu Gly Val Arg
130 135 140
Glu Asn Asp Lys Asp Leu Lys Ile Cys Val Ile Glu Asp Thr Ser Leu
145 150 155 160
Ser Gly Val Asp Asn Cys Cys Cys Gln Asp Leu Leu Gly Ile Leu Gln
165 170 175
Glu Asn Cys Ser Asp Asn Lys Arg Gly Ser Ser Ser Asn Asp Ser Cys
180 185 190
Asp Asn Lys Asn Gln Asp Glu Cys Gln Lys Lys Leu Glu Lys Val Phe
195 200 205
Ala Ser Leu Thr Asn Gly Tyr Lys Cys Asp Lys Cys Lys Ser Gly Thr
210 215 220
Ser Arg Ser Lys Lys Lys Trp Ile Trp Lys Lys Ser Ser Gly Asn Glu
225 230 235 240
Glu Gly Leu Gln Glu Glu Tyr Ala Asn Thr Ile Gly Leu Pro Pro Arg
245 250 255
Thr Gln Ser Leu Tyr Leu Gly Asn Leu Pro Lys Leu Glu Asn Val Cys
260 265 270
Glu Asp Val Lys Asp Ile Asn Phe Asp Thr Lys Glu Lys Phe Leu Ala
275 280 285
Gly Cys Leu Ile Val Ser Phe His Glu Gly Lys Asn Leu Lys Lys Arg
290 295 300
Tyr Pro Gln Asn Lys Asn Ser Gly Asn Lys Glu Asn Leu Cys Lys Ala
305 310 315 320
Leu Glu Tyr Ser Phe Ala Asp Tyr Gly Asp Leu Ile Lys Gly Thr Ser
325 330 335
Ile Trp Asp Asn Glu Tyr Thr Lys Asp Leu Glu Leu Asn Leu Gln Asn
340 345 350
Asn Phe Gly Lys Leu Phe Gly Lys Tyr Ile Lys Lys Asn Asn Thr Ala
355 360 365
Glu Gln Asp Thr Ser Tyr Ser Ser Leu Asp Glu Leu Arg Glu Ser Trp
370 375 380
Trp Asn Thr Asn Lys Lys Tyr Ile Trp Thr Ala Met Lys His Gly Ala
385 390 395 400
Glu Met Asn Ile Thr Thr Cys Asn Ala Asp Gly Ser Val Thr Gly Ser
405 410 415
Gly Ser Ser Cys Asp Asp Ile Pro Thr Ile Asp Leu Ile Pro Gln Tyr
420 425 430
Leu Arg Phe Leu Gln Glu Trp Val Glu Asn Phe Cys Glu Gln Arg Gln
435 440 445
Ala Lys Val Lys Asp Val Ile Thr Asn Cys Lys Ser Cys Lys Glu Ser
450 455 460
Gly Asn Lys Cys Lys Thr Glu Cys Lys Thr Lys Cys Lys Asp Glu Cys
465 470 475 480
Glu Lys Tyr Lys Lys Phe Ile Glu Ala Cys Gly Thr Ala Gly Gly Gly
485 490 495
Ile Gly Thr Ala Gly Ser Pro Trp Ser Lys Arg Trp Asp Gln Ile Tyr
500 505 510
Lys Arg Tyr Ser Lys His Ile Glu Asp Ala Lys Arg Asn Arg Lys Ala
515 520 525
Gly Thr Lys Asn Cys Gly Thr Ser Ser Thr Thr Asn Ala Ala Ala Ser
530 535 540
Thr Asp Glu Asn Lys Cys Val Gln Ser Asp Ile Asp Ser Phe Phe Lys
545 550 555 560
His Leu Ile Asp Ile Gly Leu Thr Thr Pro Ser Ser Tyr Leu Ser Asn
565 570 575
Val Leu Asp Asp Asn Ile Cys Gly Ala Asp Lys Ala Pro Trp Thr Thr
580 585 590
Tyr Thr Thr Tyr Thr Thr Thr Glu Lys Cys Asn Lys Glu Arg Asp Lys
595 600 605
Ser Lys Ser Gln Ser Ser Asp Thr Leu Val Val Val Asn Val Pro Ser
610 615 620
Pro Leu Gly Asn Thr Pro Tyr Arg Tyr Lys Tyr Ala Cys Gln Cys Lys
625 630 635 640
Ile Pro Thr Asn Glu Glu Thr Cys Asp Asp Arg Lys Glu Tyr Met Asn
645 650 655
Gln Trp Ser Cys Gly Ser Ala Arg Thr Met Lys Arg Gly Tyr Lys Asn
660 665 670
Asp Asn Tyr Glu Leu Cys Lys Tyr Asn Gly Val Asp Val Lys Pro Thr
675 680 685
Thr Val Arg Ser Asn Ser Ser Lys Leu Asp Pro Glu Gly Gly Ser Leu
690 695 700
Ala Ala Leu Thr Ala His Gln Ala Cys His Leu Pro Leu Glu Thr Phe
705 710 715 720
Thr Arg His Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly
725 730 735
Tyr Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser
740 745 750
Trp Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly
755 760 765
Ser Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala
770 775 780
Arg Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val Arg
785 790 795 800
Gln Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Asn Gly Pro Ala Asp
805 810 815
Ser Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu Phe
820 825 830
Leu Gly Asp Gly Gly Asp Ile Ser Phe Ser Thr Arg Gly Thr Gln Asn
835 840 845
Trp Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg
850 855 860
Gly Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln
865 870 875 880
Ser Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala
885 890 895
Ile Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly
900 905 910
Tyr Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly
915 920 925
Ala Leu Leu Arg Val Tyr Val Pro Arg Ser Ser Leu Pro Gly Phe Tyr
930 935 940
Arg Thr Ser Leu Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu Val Glu
945 950 955 960
Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala Ile Thr Gly
965 970 975
Pro Glu Glu Glu Gly Gly Arg Leu Glu Thr Ile Leu Gly Trp Pro Leu
980 985 990
Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr Asp Pro Arg
995 1000 1005
Asn Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys Glu
1010 1015 1020
Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly Lys
1025 1030 1035
Pro Pro Arg Lys Asp Glu Leu
1040 1045
<210> 62
<211> 987
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 62
Asn Tyr Ile Lys Gly Asp Pro Tyr Phe Ala Glu Tyr Ala Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Pro Ser Asp Ala Asn Asn Pro Ser Gly Glu Thr
20 25 30
Ala Asn His Asn Asp Glu Ala Cys Asn Cys Asn Glu Ser Gly Ile Ser
35 40 45
Ser Val Gly Gln Ala Gln Thr Ser Gly Pro Ser Ser Asn Lys Thr Cys
50 55 60
Ile Thr His Ser Ser Ile Lys Thr Asn Lys Lys Lys Glu Cys Lys Asp
65 70 75 80
Val Lys Leu Gly Val Arg Glu Asn Asp Lys Asp Leu Lys Ile Cys Val
85 90 95
Ile Glu Asp Thr Ser Leu Ser Gly Val Asp Asn Cys Cys Cys Gln Asp
100 105 110
Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Arg Gly Ser
115 120 125
Ser Ser Asn Asp Ser Cys Asp Asn Lys Asn Gln Asp Glu Cys Gln Lys
130 135 140
Lys Leu Glu Lys Val Phe Ala Ser Leu Thr Asn Gly Tyr Lys Cys Asp
145 150 155 160
Lys Cys Lys Ser Gly Thr Ser Arg Ser Lys Lys Lys Trp Ile Trp Lys
165 170 175
Lys Ser Ser Gly Asn Glu Glu Gly Leu Gln Glu Glu Tyr Ala Asn Thr
180 185 190
Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu Gly Asn Leu Pro
195 200 205
Lys Leu Glu Asn Val Cys Glu Asp Val Lys Asp Ile Asn Phe Asp Thr
210 215 220
Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Val Ser Phe His Glu Gly
225 230 235 240
Lys Asn Leu Lys Lys Arg Tyr Pro Gln Asn Lys Asn Ser Gly Asn Lys
245 250 255
Glu Asn Leu Cys Lys Ala Leu Glu Tyr Ser Phe Ala Asp Tyr Gly Asp
260 265 270
Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp Leu
275 280 285
Glu Leu Asn Leu Gln Asn Asn Phe Gly Lys Leu Phe Gly Lys Tyr Ile
290 295 300
Lys Lys Asn Asn Thr Ala Glu Gln Asp Thr Ser Tyr Ser Ser Leu Asp
305 310 315 320
Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Thr
325 330 335
Ala Met Lys His Gly Ala Glu Met Asn Ile Thr Thr Cys Asn Ala Asp
340 345 350
Gly Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp Ile Pro Thr Ile
355 360 365
Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu Asn
370 375 380
Phe Cys Glu Gln Arg Gln Ala Lys Val Lys Asp Val Ile Thr Asn Cys
385 390 395 400
Lys Ser Cys Lys Glu Ser Gly Asn Lys Cys Lys Thr Glu Cys Lys Thr
405 410 415
Lys Cys Lys Asp Glu Cys Glu Lys Tyr Lys Lys Phe Ile Glu Ala Cys
420 425 430
Gly Thr Ala Gly Gly Gly Ile Gly Thr Ala Gly Ser Pro Trp Ser Lys
435 440 445
Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys His Ile Glu Asp Ala
450 455 460
Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Thr Ser Ser Thr
465 470 475 480
Thr Asn Ala Ala Ala Ser Thr Asp Glu Asn Lys Cys Val Gln Ser Asp
485 490 495
Ile Asp Ser Phe Phe Lys His Leu Ile Asp Ile Gly Leu Thr Thr Pro
500 505 510
Ser Ser Tyr Leu Ser Asn Val Leu Asp Asp Asn Ile Cys Gly Ala Asp
515 520 525
Lys Ala Pro Trp Thr Thr Tyr Thr Thr Tyr Thr Thr Thr Glu Lys Cys
530 535 540
Asn Lys Glu Arg Asp Lys Ser Lys Ser Gln Ser Ser Asp Thr Leu Val
545 550 555 560
Val Val Asn Val Pro Ser Pro Leu Gly Asn Thr Pro Tyr Arg Tyr Lys
565 570 575
Tyr Ala Cys Gln Cys Lys Ile Pro Thr Asn Glu Glu Thr Cys Asp Asp
580 585 590
Arg Lys Glu Tyr Met Asn Gln Trp Ser Cys Gly Ser Ala Arg Thr Met
595 600 605
Lys Arg Gly Tyr Lys Asn Asp Asn Tyr Glu Leu Cys Lys Tyr Asn Gly
610 615 620
Val Asp Val Lys Pro Thr Thr Val Arg Ser Asn Ser Ser Lys Leu Asp
625 630 635 640
Pro Glu Gly Gly Ser Leu Ala Ala Leu Thr Ala His Gln Ala Cys His
645 650 655
Leu Pro Leu Glu Thr Phe Thr Arg His Arg Gln Pro Arg Gly Trp Glu
660 665 670
Gln Leu Glu Gln Cys Gly Tyr Pro Val Gln Arg Leu Val Ala Leu Tyr
675 680 685
Leu Ala Ala Arg Leu Ser Trp Asn Gln Val Asp Gln Val Ile Arg Asn
690 695 700
Ala Leu Ala Ser Pro Gly Ser Gly Gly Asp Leu Gly Glu Ala Ile Arg
705 710 715 720
Glu Gln Pro Glu Gln Ala Arg Leu Ala Leu Thr Leu Ala Ala Ala Glu
725 730 735
Ser Glu Arg Phe Val Arg Gln Gly Thr Gly Asn Asp Glu Ala Gly Ala
740 745 750
Ala Asn Gly Pro Ala Asp Ser Gly Asp Ala Leu Leu Glu Arg Asn Tyr
755 760 765
Pro Thr Gly Ala Glu Phe Leu Gly Asp Gly Gly Asp Ile Ser Phe Ser
770 775 780
Thr Arg Gly Thr Gln Asn Trp Thr Val Glu Arg Leu Leu Gln Ala His
785 790 795 800
Arg Gln Leu Glu Glu Arg Gly Tyr Val Phe Val Gly Tyr His Gly Thr
805 810 815
Phe Leu Glu Ala Ala Gln Ser Ile Val Phe Gly Gly Val Arg Ala Arg
820 825 830
Ser Gln Asp Leu Asp Ala Ile Trp Arg Gly Phe Tyr Ile Ala Gly Asp
835 840 845
Pro Ala Leu Ala Tyr Gly Tyr Ala Gln Asp Gln Glu Pro Asp Ala Arg
850 855 860
Gly Arg Ile Arg Asn Gly Ala Leu Leu Arg Val Tyr Val Pro Arg Ser
865 870 875 880
Ser Leu Pro Gly Phe Tyr Arg Thr Ser Leu Thr Leu Ala Ala Pro Glu
885 890 895
Ala Ala Gly Glu Val Glu Arg Leu Ile Gly His Pro Leu Pro Leu Arg
900 905 910
Leu Asp Ala Ile Thr Gly Pro Glu Glu Glu Gly Gly Arg Leu Glu Thr
915 920 925
Ile Leu Gly Trp Pro Leu Ala Glu Arg Thr Val Val Ile Pro Ser Ala
930 935 940
Ile Pro Thr Asp Pro Arg Asn Val Gly Gly Asp Leu Asp Pro Ser Ser
945 950 955 960
Ile Pro Asp Lys Glu Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser
965 970 975
Gln Pro Gly Lys Pro Pro Arg Lys Asp Glu Leu
980 985
<210> 63
<211> 871
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 63
Asn Tyr Ile Lys Gly Asp Pro Tyr Phe Ala Glu Tyr Ala Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Pro Ser Asp Ala Asn Asn Pro Ser Gly Glu Thr
20 25 30
Ala Asn His Asn Asp Glu Ala Cys Asn Cys Asn Glu Ser Gly Ile Ser
35 40 45
Ser Val Gly Gln Ala Gln Thr Ser Gly Pro Ser Ser Asn Lys Thr Cys
50 55 60
Ile Thr His Ser Ser Ile Lys Thr Asn Lys Lys Lys Glu Cys Lys Asp
65 70 75 80
Val Lys Leu Gly Val Arg Glu Asn Asp Lys Asp Leu Lys Ile Cys Val
85 90 95
Ile Glu Asp Thr Ser Leu Ser Gly Val Asp Asn Cys Cys Cys Gln Asp
100 105 110
Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Arg Gly Ser
115 120 125
Ser Ser Asn Asp Ser Cys Asp Asn Lys Asn Gln Asp Glu Cys Gln Lys
130 135 140
Lys Leu Glu Lys Val Phe Ala Ser Leu Thr Asn Gly Tyr Lys Cys Asp
145 150 155 160
Lys Cys Lys Ser Gly Thr Ser Arg Ser Lys Lys Lys Trp Ile Trp Lys
165 170 175
Lys Ser Ser Gly Asn Glu Glu Gly Leu Gln Glu Glu Tyr Ala Asn Thr
180 185 190
Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu Gly Asn Leu Pro
195 200 205
Lys Leu Glu Asn Val Cys Glu Asp Val Lys Asp Ile Asn Phe Asp Thr
210 215 220
Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Val Ser Phe His Glu Gly
225 230 235 240
Lys Asn Leu Lys Lys Arg Tyr Pro Gln Asn Lys Asn Ser Gly Asn Lys
245 250 255
Glu Asn Leu Cys Lys Ala Leu Glu Tyr Ser Phe Ala Asp Tyr Gly Asp
260 265 270
Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp Leu
275 280 285
Glu Leu Asn Leu Gln Asn Asn Phe Gly Lys Leu Phe Gly Lys Tyr Ile
290 295 300
Lys Lys Asn Asn Thr Ala Glu Gln Asp Thr Ser Tyr Ser Ser Leu Asp
305 310 315 320
Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Thr
325 330 335
Ala Met Lys His Gly Ala Glu Met Asn Ile Thr Thr Cys Asn Ala Asp
340 345 350
Gly Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp Ile Pro Thr Ile
355 360 365
Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu Asn
370 375 380
Phe Cys Glu Gln Arg Gln Ala Lys Val Lys Asp Val Ile Thr Asn Cys
385 390 395 400
Lys Ser Cys Lys Glu Ser Gly Asn Lys Cys Lys Thr Glu Cys Lys Thr
405 410 415
Lys Cys Lys Asp Glu Cys Glu Lys Tyr Lys Lys Phe Ile Glu Ala Cys
420 425 430
Gly Thr Ala Gly Gly Gly Ile Gly Thr Ala Gly Ser Pro Trp Ser Lys
435 440 445
Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys His Ile Glu Asp Ala
450 455 460
Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Thr Ser Ser Thr
465 470 475 480
Thr Asn Ala Ala Ala Ser Thr Asp Glu Asn Lys Cys Val Gln Ser Asp
485 490 495
Ile Asp Ser Phe Phe Lys His Leu Ile Asp Ile Gly Leu Thr Thr Pro
500 505 510
Ser Ser Tyr Leu Ser Asn Val Leu Asp Asp Asn Ile Cys Gly Ala Asp
515 520 525
Lys Ala Pro Trp Thr Thr Tyr Thr Thr Tyr Thr Thr Thr Glu Lys Cys
530 535 540
Asn Lys Glu Arg Asp Lys Ser Lys Ser Gln Ser Ser Asp Thr Leu Val
545 550 555 560
Val Val Asn Val Pro Ser Pro Leu Gly Asn Thr Pro Tyr Arg Tyr Lys
565 570 575
Tyr Ala Cys Gln Cys Lys Ile Pro Thr Asn Glu Glu Thr Cys Asp Asp
580 585 590
Arg Lys Glu Tyr Met Asn Gln Trp Ser Cys Gly Ser Ala Arg Thr Met
595 600 605
Lys Arg Gly Tyr Lys Asn Asp Asn Tyr Glu Leu Cys Lys Tyr Asn Gly
610 615 620
Val Asp Val Lys Pro Thr Thr Val Arg Ser Asn Ser Ser Lys Leu Asp
625 630 635 640
Arg His Arg Gln Pro Arg Gly Trp Glu Gln Leu Tyr Pro Thr Gly Ala
645 650 655
Glu Phe Leu Gly Asp Gly Gly Asp Ile Ser Phe Ser Thr Arg Gly Thr
660 665 670
Gln Asn Trp Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu
675 680 685
Glu Arg Gly Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala
690 695 700
Ala Gln Ser Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu
705 710 715 720
Asp Ala Ile Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala
725 730 735
Tyr Gly Tyr Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg
740 745 750
Asn Gly Ala Leu Leu Arg Val Tyr Val Pro Arg Ser Ser Leu Pro Gly
755 760 765
Phe Tyr Arg Thr Ser Leu Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu
770 775 780
Val Glu Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala Ile
785 790 795 800
Thr Gly Pro Glu Glu Glu Gly Gly Arg Leu Glu Thr Ile Leu Gly Trp
805 810 815
Pro Leu Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr Asp
820 825 830
Pro Arg Asn Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys
835 840 845
Glu Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly Lys
850 855 860
Pro Pro Arg Lys Asp Glu Leu
865 870
<210> 64
<211> 1258
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 64
Arg Pro Asp Phe Cys Leu Glu Pro Pro Tyr Thr Gly Pro Cys Lys Ala
1 5 10 15
Arg Ile Ile Arg Tyr Phe Tyr Asn Ala Lys Ala Gly Leu Cys Gln Thr
20 25 30
Phe Val Tyr Gly Gly Cys Arg Ala Lys Arg Asn Asn Phe Lys Ser Ala
35 40 45
Glu Asp Cys Met Arg Thr Cys Gly Gly Ala Asn His Ser Asp Ser Gly
50 55 60
Lys Tyr Asp Pro Cys Glu Lys Lys Leu Pro Pro Tyr Asp Asp Asn Asp
65 70 75 80
Gln Trp Lys Cys Gln Gln Asn Ser Ser Asp Gly Ser Gly Lys Pro Glu
85 90 95
Asn Ile Cys Val Pro Pro Arg Arg Glu Arg Leu Cys Thr Tyr Asn Leu
100 105 110
Glu Asn Leu Lys Phe Asp Lys Ile Arg Asp Asn Asn Ala Phe Leu Ala
115 120 125
Asp Val Leu Leu Thr Ala Arg Asn Glu Gly Glu Lys Ile Val Gln Asn
130 135 140
His Pro Asp Thr Asn Ser Ser Asn Val Cys Asn Ala Leu Glu Arg Ser
145 150 155 160
Phe Ala Asp Leu Ala Asp Ile Ile Arg Gly Thr Asp Gln Trp Lys Gly
165 170 175
Thr Asn Ser Asn Leu Glu Lys Asn Leu Lys Gln Met Phe Ala Lys Ile
180 185 190
Arg Glu Asn Asp Lys Val Leu Gln Asp Lys Tyr Pro Lys Asp Gln Lys
195 200 205
Tyr Thr Lys Leu Arg Glu Ala Trp Trp Asn Ala Asn Arg Gln Lys Val
210 215 220
Trp Glu Val Ile Thr Cys Gly Ala Arg Ser Asn Asp Leu Leu Ile Lys
225 230 235 240
Arg Gly Trp Arg Thr Ser Gly Lys Ser Asp Arg Lys Lys Asn Phe Glu
245 250 255
Leu Cys Arg Lys Cys Gly His Tyr Glu Lys Glu Val Pro Thr Lys Leu
260 265 270
Asp Tyr Val Pro Gln Phe Leu Arg Trp Leu Thr Glu Trp Ile Glu Asp
275 280 285
Phe Tyr Arg Glu Lys Gln Asn Leu Ile Asp Asp Met Glu Arg His Arg
290 295 300
Glu Glu Cys Thr Arg Glu Asp His Lys Ser Lys Glu Gly Thr Ser Tyr
305 310 315 320
Cys Ser Thr Cys Lys Asp Lys Cys Lys Lys Tyr Cys Glu Cys Val Lys
325 330 335
Lys Trp Lys Thr Glu Trp Glu Asn Gln Glu Asn Lys Tyr Lys Asp Leu
340 345 350
Tyr Glu Gln Asn Lys Asn Lys Thr Ser Gln Lys Asn Thr Ser Arg Tyr
355 360 365
Asp Asp Tyr Val Lys Asp Phe Phe Glu Lys Leu Glu Ala Asn Tyr Ser
370 375 380
Ser Leu Glu Asn Tyr Ile Lys Gly Asp Pro Tyr Phe Ala Glu Tyr Ala
385 390 395 400
Thr Lys Leu Ser Phe Ile Leu Asn Pro Ser Asp Ala Asn Asn Pro Ser
405 410 415
Gly Glu Thr Ala Asn His Asn Asp Glu Ala Cys Asn Cys Asn Glu Ser
420 425 430
Gly Ile Ser Ser Val Gly Gln Ala Gln Thr Ser Gly Pro Ser Ser Asn
435 440 445
Lys Thr Cys Ile Thr His Ser Ser Ile Lys Thr Asn Lys Lys Lys Glu
450 455 460
Cys Lys Asp Val Lys Leu Gly Val Arg Glu Asn Asp Lys Asp Leu Lys
465 470 475 480
Ile Cys Val Ile Glu Asp Thr Ser Leu Ser Gly Val Asp Asn Cys Cys
485 490 495
Cys Gln Asp Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys
500 505 510
Arg Gly Ser Ser Ser Asn Asp Ser Cys Asp Asn Lys Asn Gln Asp Glu
515 520 525
Cys Gln Lys Lys Leu Glu Lys Val Phe Ala Ser Leu Thr Asn Gly Tyr
530 535 540
Lys Cys Asp Lys Cys Lys Ser Gly Thr Ser Arg Ser Lys Lys Lys Trp
545 550 555 560
Ile Trp Lys Lys Ser Ser Gly Asn Glu Glu Gly Leu Gln Glu Glu Tyr
565 570 575
Ala Asn Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu Gly
580 585 590
Asn Leu Pro Lys Leu Glu Asn Val Cys Glu Asp Val Lys Asp Ile Asn
595 600 605
Phe Asp Thr Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Val Ser Phe
610 615 620
His Glu Gly Lys Asn Leu Lys Lys Arg Tyr Pro Gln Asn Lys Asn Ser
625 630 635 640
Gly Asn Lys Glu Asn Leu Cys Lys Ala Leu Glu Tyr Ser Phe Ala Asp
645 650 655
Tyr Gly Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr
660 665 670
Lys Asp Leu Glu Leu Asn Leu Gln Asn Asn Phe Gly Lys Leu Phe Gly
675 680 685
Lys Tyr Ile Lys Lys Asn Asn Thr Ala Glu Gln Asp Thr Ser Tyr Ser
690 695 700
Ser Leu Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr
705 710 715 720
Ile Trp Thr Ala Met Lys His Gly Ala Glu Met Asn Ile Thr Thr Cys
725 730 735
Asn Ala Asp Gly Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp Ile
740 745 750
Pro Thr Ile Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp
755 760 765
Val Glu Asn Phe Cys Glu Gln Arg Gln Ala Lys Val Lys Asp Val Ile
770 775 780
Thr Asn Cys Lys Ser Cys Lys Glu Ser Gly Asn Lys Cys Lys Thr Glu
785 790 795 800
Cys Lys Thr Lys Cys Lys Asp Glu Cys Glu Lys Tyr Lys Lys Phe Ile
805 810 815
Glu Ala Cys Gly Thr Ala Gly Gly Gly Ile Gly Thr Ala Gly Ser Pro
820 825 830
Trp Ser Lys Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys His Ile
835 840 845
Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Thr
850 855 860
Ser Ser Thr Thr Asn Ala Ala Ala Ser Thr Asp Glu Asn Lys Cys Val
865 870 875 880
Gln Ser Asp Ile Asp Ser Phe Phe Lys His Leu Ile Asp Ile Gly Leu
885 890 895
Thr Thr Pro Ser Ser Tyr Leu Ser Asn Val Leu Asp Asp Asn Ile Cys
900 905 910
Gly Ala Asp Lys Ala Pro Trp Thr Thr Tyr Thr Thr Tyr Thr Thr Thr
915 920 925
Glu Lys Cys Asn Lys Glu Arg Asp Lys Ser Lys Ser Gln Ser Ser Asp
930 935 940
Thr Leu Val Val Val Asn Val Pro Ser Pro Leu Gly Asn Thr Pro Tyr
945 950 955 960
Arg Tyr Lys Tyr Ala Cys Gln Cys Lys Ile Pro Thr Asn Glu Glu Thr
965 970 975
Cys Asp Asp Arg Lys Glu Tyr Met Asn Gln Trp Ser Cys Gly Ser Ala
980 985 990
Arg Thr Met Lys Arg Gly Tyr Lys Asn Asp Asn Tyr Glu Leu Cys Lys
995 1000 1005
Tyr Asn Gly Val Asp Val Lys Pro Thr Thr Val Arg Ser Asn Ser
1010 1015 1020
Ser Lys Leu Asp Arg His Arg Gln Pro Arg Gly Trp Glu Gln Leu
1025 1030 1035
Tyr Pro Thr Gly Ala Glu Phe Leu Gly Asp Gly Gly Asp Ile Ser
1040 1045 1050
Phe Ser Thr Arg Gly Thr Gln Asn Trp Thr Val Glu Arg Leu Leu
1055 1060 1065
Gln Ala His Arg Gln Leu Glu Glu Arg Gly Tyr Val Phe Val Gly
1070 1075 1080
Tyr His Gly Thr Phe Leu Glu Ala Ala Gln Ser Ile Val Phe Gly
1085 1090 1095
Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala Ile Trp Arg Gly
1100 1105 1110
Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly Tyr Ala Gln
1115 1120 1125
Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly Ala Leu
1130 1135 1140
Leu Arg Val Tyr Val Pro Arg Ser Ser Leu Pro Gly Phe Tyr Arg
1145 1150 1155
Thr Ser Leu Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu Val Glu
1160 1165 1170
Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala Ile Thr
1175 1180 1185
Gly Pro Glu Glu Glu Gly Gly Arg Leu Glu Thr Ile Leu Gly Trp
1190 1195 1200
Pro Leu Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr
1205 1210 1215
Asp Pro Arg Asn Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro
1220 1225 1230
Asp Lys Glu Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln
1235 1240 1245
Pro Gly Lys Pro Pro Arg Lys Asp Glu Leu
1250 1255
<210> 65
<211> 1374
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 65
Arg Pro Asp Phe Cys Leu Glu Pro Pro Tyr Thr Gly Pro Cys Lys Ala
1 5 10 15
Arg Ile Ile Arg Tyr Phe Tyr Asn Ala Lys Ala Gly Leu Cys Gln Thr
20 25 30
Phe Val Tyr Gly Gly Cys Arg Ala Lys Arg Asn Asn Phe Lys Ser Ala
35 40 45
Glu Asp Cys Met Arg Thr Cys Gly Gly Ala Asn His Ser Asp Ser Gly
50 55 60
Lys Tyr Asp Pro Cys Glu Lys Lys Leu Pro Pro Tyr Asp Asp Asn Asp
65 70 75 80
Gln Trp Lys Cys Gln Gln Asn Ser Ser Asp Gly Ser Gly Lys Pro Glu
85 90 95
Asn Ile Cys Val Pro Pro Arg Arg Glu Arg Leu Cys Thr Tyr Asn Leu
100 105 110
Glu Asn Leu Lys Phe Asp Lys Ile Arg Asp Asn Asn Ala Phe Leu Ala
115 120 125
Asp Val Leu Leu Thr Ala Arg Asn Glu Gly Glu Lys Ile Val Gln Asn
130 135 140
His Pro Asp Thr Asn Ser Ser Asn Val Cys Asn Ala Leu Glu Arg Ser
145 150 155 160
Phe Ala Asp Leu Ala Asp Ile Ile Arg Gly Thr Asp Gln Trp Lys Gly
165 170 175
Thr Asn Ser Asn Leu Glu Lys Asn Leu Lys Gln Met Phe Ala Lys Ile
180 185 190
Arg Glu Asn Asp Lys Val Leu Gln Asp Lys Tyr Pro Lys Asp Gln Lys
195 200 205
Tyr Thr Lys Leu Arg Glu Ala Trp Trp Asn Ala Asn Arg Gln Lys Val
210 215 220
Trp Glu Val Ile Thr Cys Gly Ala Arg Ser Asn Asp Leu Leu Ile Lys
225 230 235 240
Arg Gly Trp Arg Thr Ser Gly Lys Ser Asp Arg Lys Lys Asn Phe Glu
245 250 255
Leu Cys Arg Lys Cys Gly His Tyr Glu Lys Glu Val Pro Thr Lys Leu
260 265 270
Asp Tyr Val Pro Gln Phe Leu Arg Trp Leu Thr Glu Trp Ile Glu Asp
275 280 285
Phe Tyr Arg Glu Lys Gln Asn Leu Ile Asp Asp Met Glu Arg His Arg
290 295 300
Glu Glu Cys Thr Arg Glu Asp His Lys Ser Lys Glu Gly Thr Ser Tyr
305 310 315 320
Cys Ser Thr Cys Lys Asp Lys Cys Lys Lys Tyr Cys Glu Cys Val Lys
325 330 335
Lys Trp Lys Thr Glu Trp Glu Asn Gln Glu Asn Lys Tyr Lys Asp Leu
340 345 350
Tyr Glu Gln Asn Lys Asn Lys Thr Ser Gln Lys Asn Thr Ser Arg Tyr
355 360 365
Asp Asp Tyr Val Lys Asp Phe Phe Glu Lys Leu Glu Ala Asn Tyr Ser
370 375 380
Ser Leu Glu Asn Tyr Ile Lys Gly Asp Pro Tyr Phe Ala Glu Tyr Ala
385 390 395 400
Thr Lys Leu Ser Phe Ile Leu Asn Pro Ser Asp Ala Asn Asn Pro Ser
405 410 415
Gly Glu Thr Ala Asn His Asn Asp Glu Ala Cys Asn Cys Asn Glu Ser
420 425 430
Gly Ile Ser Ser Val Gly Gln Ala Gln Thr Ser Gly Pro Ser Ser Asn
435 440 445
Lys Thr Cys Ile Thr His Ser Ser Ile Lys Thr Asn Lys Lys Lys Glu
450 455 460
Cys Lys Asp Val Lys Leu Gly Val Arg Glu Asn Asp Lys Asp Leu Lys
465 470 475 480
Ile Cys Val Ile Glu Asp Thr Ser Leu Ser Gly Val Asp Asn Cys Cys
485 490 495
Cys Gln Asp Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys
500 505 510
Arg Gly Ser Ser Ser Asn Asp Ser Cys Asp Asn Lys Asn Gln Asp Glu
515 520 525
Cys Gln Lys Lys Leu Glu Lys Val Phe Ala Ser Leu Thr Asn Gly Tyr
530 535 540
Lys Cys Asp Lys Cys Lys Ser Gly Thr Ser Arg Ser Lys Lys Lys Trp
545 550 555 560
Ile Trp Lys Lys Ser Ser Gly Asn Glu Glu Gly Leu Gln Glu Glu Tyr
565 570 575
Ala Asn Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu Gly
580 585 590
Asn Leu Pro Lys Leu Glu Asn Val Cys Glu Asp Val Lys Asp Ile Asn
595 600 605
Phe Asp Thr Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Val Ser Phe
610 615 620
His Glu Gly Lys Asn Leu Lys Lys Arg Tyr Pro Gln Asn Lys Asn Ser
625 630 635 640
Gly Asn Lys Glu Asn Leu Cys Lys Ala Leu Glu Tyr Ser Phe Ala Asp
645 650 655
Tyr Gly Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr
660 665 670
Lys Asp Leu Glu Leu Asn Leu Gln Asn Asn Phe Gly Lys Leu Phe Gly
675 680 685
Lys Tyr Ile Lys Lys Asn Asn Thr Ala Glu Gln Asp Thr Ser Tyr Ser
690 695 700
Ser Leu Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr
705 710 715 720
Ile Trp Thr Ala Met Lys His Gly Ala Glu Met Asn Ile Thr Thr Cys
725 730 735
Asn Ala Asp Gly Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp Ile
740 745 750
Pro Thr Ile Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp
755 760 765
Val Glu Asn Phe Cys Glu Gln Arg Gln Ala Lys Val Lys Asp Val Ile
770 775 780
Thr Asn Cys Lys Ser Cys Lys Glu Ser Gly Asn Lys Cys Lys Thr Glu
785 790 795 800
Cys Lys Thr Lys Cys Lys Asp Glu Cys Glu Lys Tyr Lys Lys Phe Ile
805 810 815
Glu Ala Cys Gly Thr Ala Gly Gly Gly Ile Gly Thr Ala Gly Ser Pro
820 825 830
Trp Ser Lys Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys His Ile
835 840 845
Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Thr
850 855 860
Ser Ser Thr Thr Asn Ala Ala Ala Ser Thr Asp Glu Asn Lys Cys Val
865 870 875 880
Gln Ser Asp Ile Asp Ser Phe Phe Lys His Leu Ile Asp Ile Gly Leu
885 890 895
Thr Thr Pro Ser Ser Tyr Leu Ser Asn Val Leu Asp Asp Asn Ile Cys
900 905 910
Gly Ala Asp Lys Ala Pro Trp Thr Thr Tyr Thr Thr Tyr Thr Thr Thr
915 920 925
Glu Lys Cys Asn Lys Glu Arg Asp Lys Ser Lys Ser Gln Ser Ser Asp
930 935 940
Thr Leu Val Val Val Asn Val Pro Ser Pro Leu Gly Asn Thr Pro Tyr
945 950 955 960
Arg Tyr Lys Tyr Ala Cys Gln Cys Lys Ile Pro Thr Asn Glu Glu Thr
965 970 975
Cys Asp Asp Arg Lys Glu Tyr Met Asn Gln Trp Ser Cys Gly Ser Ala
980 985 990
Arg Thr Met Lys Arg Gly Tyr Lys Asn Asp Asn Tyr Glu Leu Cys Lys
995 1000 1005
Tyr Asn Gly Val Asp Val Lys Pro Thr Thr Val Arg Ser Asn Ser
1010 1015 1020
Ser Lys Leu Asp Pro Glu Gly Gly Ser Leu Ala Ala Leu Thr Ala
1025 1030 1035
His Gln Ala Cys His Leu Pro Leu Glu Thr Phe Thr Arg His Arg
1040 1045 1050
Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly Tyr Pro Val
1055 1060 1065
Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser Trp Asn
1070 1075 1080
Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly Ser
1085 1090 1095
Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala
1100 1105 1110
Arg Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val
1115 1120 1125
Arg Gln Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Asn Gly Pro
1130 1135 1140
Ala Asp Ser Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly
1145 1150 1155
Ala Glu Phe Leu Gly Asp Gly Gly Asp Ile Ser Phe Ser Thr Arg
1160 1165 1170
Gly Thr Gln Asn Trp Thr Val Glu Arg Leu Leu Gln Ala His Arg
1175 1180 1185
Gln Leu Glu Glu Arg Gly Tyr Val Phe Val Gly Tyr His Gly Thr
1190 1195 1200
Phe Leu Glu Ala Ala Gln Ser Ile Val Phe Gly Gly Val Arg Ala
1205 1210 1215
Arg Ser Gln Asp Leu Asp Ala Ile Trp Arg Gly Phe Tyr Ile Ala
1220 1225 1230
Gly Asp Pro Ala Leu Ala Tyr Gly Tyr Ala Gln Asp Gln Glu Pro
1235 1240 1245
Asp Ala Arg Gly Arg Ile Arg Asn Gly Ala Leu Leu Arg Val Tyr
1250 1255 1260
Val Pro Arg Ser Ser Leu Pro Gly Phe Tyr Arg Thr Ser Leu Thr
1265 1270 1275
Leu Ala Ala Pro Glu Ala Ala Gly Glu Val Glu Arg Leu Ile Gly
1280 1285 1290
His Pro Leu Pro Leu Arg Leu Asp Ala Ile Thr Gly Pro Glu Glu
1295 1300 1305
Glu Gly Gly Arg Leu Glu Thr Ile Leu Gly Trp Pro Leu Ala Glu
1310 1315 1320
Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr Asp Pro Arg Asn
1325 1330 1335
Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys Glu Gln
1340 1345 1350
Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly Lys Pro
1355 1360 1365
Pro Arg Lys Asp Glu Leu
1370
<210> 66
<211> 1200
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 66
Asn His Ser Asp Ser Gly Lys Tyr Asp Pro Cys Glu Lys Lys Leu Pro
1 5 10 15
Pro Tyr Asp Asp Asn Asp Gln Trp Lys Cys Gln Gln Asn Ser Ser Asp
20 25 30
Gly Ser Gly Lys Pro Glu Asn Ile Cys Val Pro Pro Arg Arg Glu Arg
35 40 45
Leu Cys Thr Tyr Asn Leu Glu Asn Leu Lys Phe Asp Lys Ile Arg Asp
50 55 60
Asn Asn Ala Phe Leu Ala Asp Val Leu Leu Thr Ala Arg Asn Glu Gly
65 70 75 80
Glu Lys Ile Val Gln Asn His Pro Asp Thr Asn Ser Ser Asn Val Cys
85 90 95
Asn Ala Leu Glu Arg Ser Phe Ala Asp Leu Ala Asp Ile Ile Arg Gly
100 105 110
Thr Asp Gln Trp Lys Gly Thr Asn Ser Asn Leu Glu Lys Asn Leu Lys
115 120 125
Gln Met Phe Ala Lys Ile Arg Glu Asn Asp Lys Val Leu Gln Asp Lys
130 135 140
Tyr Pro Lys Asp Gln Lys Tyr Thr Lys Leu Arg Glu Ala Trp Trp Asn
145 150 155 160
Ala Asn Arg Gln Lys Val Trp Glu Val Ile Thr Cys Gly Ala Arg Ser
165 170 175
Asn Asp Leu Leu Ile Lys Arg Gly Trp Arg Thr Ser Gly Lys Ser Asp
180 185 190
Arg Lys Lys Asn Phe Glu Leu Cys Arg Lys Cys Gly His Tyr Glu Lys
195 200 205
Glu Val Pro Thr Lys Leu Asp Tyr Val Pro Gln Phe Leu Arg Trp Leu
210 215 220
Thr Glu Trp Ile Glu Asp Phe Tyr Arg Glu Lys Gln Asn Leu Ile Asp
225 230 235 240
Asp Met Glu Arg His Arg Glu Glu Cys Thr Arg Glu Asp His Lys Ser
245 250 255
Lys Glu Gly Thr Ser Tyr Cys Ser Thr Cys Lys Asp Lys Cys Lys Lys
260 265 270
Tyr Cys Glu Cys Val Lys Lys Trp Lys Thr Glu Trp Glu Asn Gln Glu
275 280 285
Asn Lys Tyr Lys Asp Leu Tyr Glu Gln Asn Lys Asn Lys Thr Ser Gln
290 295 300
Lys Asn Thr Ser Arg Tyr Asp Asp Tyr Val Lys Asp Phe Phe Glu Lys
305 310 315 320
Leu Glu Ala Asn Tyr Ser Ser Leu Glu Asn Tyr Ile Lys Gly Asp Pro
325 330 335
Tyr Phe Ala Glu Tyr Ala Thr Lys Leu Ser Phe Ile Leu Asn Pro Ser
340 345 350
Asp Ala Asn Asn Pro Ser Gly Glu Thr Ala Asn His Asn Asp Glu Ala
355 360 365
Cys Asn Cys Asn Glu Ser Gly Ile Ser Ser Val Gly Gln Ala Gln Thr
370 375 380
Ser Gly Pro Ser Ser Asn Lys Thr Cys Ile Thr His Ser Ser Ile Lys
385 390 395 400
Thr Asn Lys Lys Lys Glu Cys Lys Asp Val Lys Leu Gly Val Arg Glu
405 410 415
Asn Asp Lys Asp Leu Lys Ile Cys Val Ile Glu Asp Thr Ser Leu Ser
420 425 430
Gly Val Asp Asn Cys Cys Cys Gln Asp Leu Leu Gly Ile Leu Gln Glu
435 440 445
Asn Cys Ser Asp Asn Lys Arg Gly Ser Ser Ser Asn Asp Ser Cys Asp
450 455 460
Asn Lys Asn Gln Asp Glu Cys Gln Lys Lys Leu Glu Lys Val Phe Ala
465 470 475 480
Ser Leu Thr Asn Gly Tyr Lys Cys Asp Lys Cys Lys Ser Gly Thr Ser
485 490 495
Arg Ser Lys Lys Lys Trp Ile Trp Lys Lys Ser Ser Gly Asn Glu Glu
500 505 510
Gly Leu Gln Glu Glu Tyr Ala Asn Thr Ile Gly Leu Pro Pro Arg Thr
515 520 525
Gln Ser Leu Tyr Leu Gly Asn Leu Pro Lys Leu Glu Asn Val Cys Glu
530 535 540
Asp Val Lys Asp Ile Asn Phe Asp Thr Lys Glu Lys Phe Leu Ala Gly
545 550 555 560
Cys Leu Ile Val Ser Phe His Glu Gly Lys Asn Leu Lys Lys Arg Tyr
565 570 575
Pro Gln Asn Lys Asn Ser Gly Asn Lys Glu Asn Leu Cys Lys Ala Leu
580 585 590
Glu Tyr Ser Phe Ala Asp Tyr Gly Asp Leu Ile Lys Gly Thr Ser Ile
595 600 605
Trp Asp Asn Glu Tyr Thr Lys Asp Leu Glu Leu Asn Leu Gln Asn Asn
610 615 620
Phe Gly Lys Leu Phe Gly Lys Tyr Ile Lys Lys Asn Asn Thr Ala Glu
625 630 635 640
Gln Asp Thr Ser Tyr Ser Ser Leu Asp Glu Leu Arg Glu Ser Trp Trp
645 650 655
Asn Thr Asn Lys Lys Tyr Ile Trp Thr Ala Met Lys His Gly Ala Glu
660 665 670
Met Asn Ile Thr Thr Cys Asn Ala Asp Gly Ser Val Thr Gly Ser Gly
675 680 685
Ser Ser Cys Asp Asp Ile Pro Thr Ile Asp Leu Ile Pro Gln Tyr Leu
690 695 700
Arg Phe Leu Gln Glu Trp Val Glu Asn Phe Cys Glu Gln Arg Gln Ala
705 710 715 720
Lys Val Lys Asp Val Ile Thr Asn Cys Lys Ser Cys Lys Glu Ser Gly
725 730 735
Asn Lys Cys Lys Thr Glu Cys Lys Thr Lys Cys Lys Asp Glu Cys Glu
740 745 750
Lys Tyr Lys Lys Phe Ile Glu Ala Cys Gly Thr Ala Gly Gly Gly Ile
755 760 765
Gly Thr Ala Gly Ser Pro Trp Ser Lys Arg Trp Asp Gln Ile Tyr Lys
770 775 780
Arg Tyr Ser Lys His Ile Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly
785 790 795 800
Thr Lys Asn Cys Gly Thr Ser Ser Thr Thr Asn Ala Ala Ala Ser Thr
805 810 815
Asp Glu Asn Lys Cys Val Gln Ser Asp Ile Asp Ser Phe Phe Lys His
820 825 830
Leu Ile Asp Ile Gly Leu Thr Thr Pro Ser Ser Tyr Leu Ser Asn Val
835 840 845
Leu Asp Asp Asn Ile Cys Gly Ala Asp Lys Ala Pro Trp Thr Thr Tyr
850 855 860
Thr Thr Tyr Thr Thr Thr Glu Lys Cys Asn Lys Glu Arg Asp Lys Ser
865 870 875 880
Lys Ser Gln Ser Ser Asp Thr Leu Val Val Val Asn Val Pro Ser Pro
885 890 895
Leu Gly Asn Thr Pro Tyr Arg Tyr Lys Tyr Ala Cys Gln Cys Lys Ile
900 905 910
Pro Thr Asn Glu Glu Thr Cys Asp Asp Arg Lys Glu Tyr Met Asn Gln
915 920 925
Trp Ser Cys Gly Ser Ala Arg Thr Met Lys Arg Gly Tyr Lys Asn Asp
930 935 940
Asn Tyr Glu Leu Cys Lys Tyr Asn Gly Val Asp Val Lys Pro Thr Thr
945 950 955 960
Val Arg Ser Asn Ser Ser Lys Leu Asp Arg His Arg Gln Pro Arg Gly
965 970 975
Trp Glu Gln Leu Tyr Pro Thr Gly Ala Glu Phe Leu Gly Asp Gly Gly
980 985 990
Asp Ile Ser Phe Ser Thr Arg Gly Thr Gln Asn Trp Thr Val Glu Arg
995 1000 1005
Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg Gly Tyr Val Phe
1010 1015 1020
Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln Ser Ile Val
1025 1030 1035
Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala Ile Trp
1040 1045 1050
Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly Tyr
1055 1060 1065
Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly
1070 1075 1080
Ala Leu Leu Arg Val Tyr Val Pro Arg Ser Ser Leu Pro Gly Phe
1085 1090 1095
Tyr Arg Thr Ser Leu Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu
1100 1105 1110
Val Glu Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala
1115 1120 1125
Ile Thr Gly Pro Glu Glu Glu Gly Gly Arg Leu Glu Thr Ile Leu
1130 1135 1140
Gly Trp Pro Leu Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile
1145 1150 1155
Pro Thr Asp Pro Arg Asn Val Gly Gly Asp Leu Asp Pro Ser Ser
1160 1165 1170
Ile Pro Asp Lys Glu Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala
1175 1180 1185
Ser Gln Pro Gly Lys Pro Pro Arg Lys Asp Glu Leu
1190 1195 1200
<210> 67
<211> 1316
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 67
Asn His Ser Asp Ser Gly Lys Tyr Asp Pro Cys Glu Lys Lys Leu Pro
1 5 10 15
Pro Tyr Asp Asp Asn Asp Gln Trp Lys Cys Gln Gln Asn Ser Ser Asp
20 25 30
Gly Ser Gly Lys Pro Glu Asn Ile Cys Val Pro Pro Arg Arg Glu Arg
35 40 45
Leu Cys Thr Tyr Asn Leu Glu Asn Leu Lys Phe Asp Lys Ile Arg Asp
50 55 60
Asn Asn Ala Phe Leu Ala Asp Val Leu Leu Thr Ala Arg Asn Glu Gly
65 70 75 80
Glu Lys Ile Val Gln Asn His Pro Asp Thr Asn Ser Ser Asn Val Cys
85 90 95
Asn Ala Leu Glu Arg Ser Phe Ala Asp Leu Ala Asp Ile Ile Arg Gly
100 105 110
Thr Asp Gln Trp Lys Gly Thr Asn Ser Asn Leu Glu Lys Asn Leu Lys
115 120 125
Gln Met Phe Ala Lys Ile Arg Glu Asn Asp Lys Val Leu Gln Asp Lys
130 135 140
Tyr Pro Lys Asp Gln Lys Tyr Thr Lys Leu Arg Glu Ala Trp Trp Asn
145 150 155 160
Ala Asn Arg Gln Lys Val Trp Glu Val Ile Thr Cys Gly Ala Arg Ser
165 170 175
Asn Asp Leu Leu Ile Lys Arg Gly Trp Arg Thr Ser Gly Lys Ser Asp
180 185 190
Arg Lys Lys Asn Phe Glu Leu Cys Arg Lys Cys Gly His Tyr Glu Lys
195 200 205
Glu Val Pro Thr Lys Leu Asp Tyr Val Pro Gln Phe Leu Arg Trp Leu
210 215 220
Thr Glu Trp Ile Glu Asp Phe Tyr Arg Glu Lys Gln Asn Leu Ile Asp
225 230 235 240
Asp Met Glu Arg His Arg Glu Glu Cys Thr Arg Glu Asp His Lys Ser
245 250 255
Lys Glu Gly Thr Ser Tyr Cys Ser Thr Cys Lys Asp Lys Cys Lys Lys
260 265 270
Tyr Cys Glu Cys Val Lys Lys Trp Lys Thr Glu Trp Glu Asn Gln Glu
275 280 285
Asn Lys Tyr Lys Asp Leu Tyr Glu Gln Asn Lys Asn Lys Thr Ser Gln
290 295 300
Lys Asn Thr Ser Arg Tyr Asp Asp Tyr Val Lys Asp Phe Phe Glu Lys
305 310 315 320
Leu Glu Ala Asn Tyr Ser Ser Leu Glu Asn Tyr Ile Lys Gly Asp Pro
325 330 335
Tyr Phe Ala Glu Tyr Ala Thr Lys Leu Ser Phe Ile Leu Asn Pro Ser
340 345 350
Asp Ala Asn Asn Pro Ser Gly Glu Thr Ala Asn His Asn Asp Glu Ala
355 360 365
Cys Asn Cys Asn Glu Ser Gly Ile Ser Ser Val Gly Gln Ala Gln Thr
370 375 380
Ser Gly Pro Ser Ser Asn Lys Thr Cys Ile Thr His Ser Ser Ile Lys
385 390 395 400
Thr Asn Lys Lys Lys Glu Cys Lys Asp Val Lys Leu Gly Val Arg Glu
405 410 415
Asn Asp Lys Asp Leu Lys Ile Cys Val Ile Glu Asp Thr Ser Leu Ser
420 425 430
Gly Val Asp Asn Cys Cys Cys Gln Asp Leu Leu Gly Ile Leu Gln Glu
435 440 445
Asn Cys Ser Asp Asn Lys Arg Gly Ser Ser Ser Asn Asp Ser Cys Asp
450 455 460
Asn Lys Asn Gln Asp Glu Cys Gln Lys Lys Leu Glu Lys Val Phe Ala
465 470 475 480
Ser Leu Thr Asn Gly Tyr Lys Cys Asp Lys Cys Lys Ser Gly Thr Ser
485 490 495
Arg Ser Lys Lys Lys Trp Ile Trp Lys Lys Ser Ser Gly Asn Glu Glu
500 505 510
Gly Leu Gln Glu Glu Tyr Ala Asn Thr Ile Gly Leu Pro Pro Arg Thr
515 520 525
Gln Ser Leu Tyr Leu Gly Asn Leu Pro Lys Leu Glu Asn Val Cys Glu
530 535 540
Asp Val Lys Asp Ile Asn Phe Asp Thr Lys Glu Lys Phe Leu Ala Gly
545 550 555 560
Cys Leu Ile Val Ser Phe His Glu Gly Lys Asn Leu Lys Lys Arg Tyr
565 570 575
Pro Gln Asn Lys Asn Ser Gly Asn Lys Glu Asn Leu Cys Lys Ala Leu
580 585 590
Glu Tyr Ser Phe Ala Asp Tyr Gly Asp Leu Ile Lys Gly Thr Ser Ile
595 600 605
Trp Asp Asn Glu Tyr Thr Lys Asp Leu Glu Leu Asn Leu Gln Asn Asn
610 615 620
Phe Gly Lys Leu Phe Gly Lys Tyr Ile Lys Lys Asn Asn Thr Ala Glu
625 630 635 640
Gln Asp Thr Ser Tyr Ser Ser Leu Asp Glu Leu Arg Glu Ser Trp Trp
645 650 655
Asn Thr Asn Lys Lys Tyr Ile Trp Thr Ala Met Lys His Gly Ala Glu
660 665 670
Met Asn Ile Thr Thr Cys Asn Ala Asp Gly Ser Val Thr Gly Ser Gly
675 680 685
Ser Ser Cys Asp Asp Ile Pro Thr Ile Asp Leu Ile Pro Gln Tyr Leu
690 695 700
Arg Phe Leu Gln Glu Trp Val Glu Asn Phe Cys Glu Gln Arg Gln Ala
705 710 715 720
Lys Val Lys Asp Val Ile Thr Asn Cys Lys Ser Cys Lys Glu Ser Gly
725 730 735
Asn Lys Cys Lys Thr Glu Cys Lys Thr Lys Cys Lys Asp Glu Cys Glu
740 745 750
Lys Tyr Lys Lys Phe Ile Glu Ala Cys Gly Thr Ala Gly Gly Gly Ile
755 760 765
Gly Thr Ala Gly Ser Pro Trp Ser Lys Arg Trp Asp Gln Ile Tyr Lys
770 775 780
Arg Tyr Ser Lys His Ile Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly
785 790 795 800
Thr Lys Asn Cys Gly Thr Ser Ser Thr Thr Asn Ala Ala Ala Ser Thr
805 810 815
Asp Glu Asn Lys Cys Val Gln Ser Asp Ile Asp Ser Phe Phe Lys His
820 825 830
Leu Ile Asp Ile Gly Leu Thr Thr Pro Ser Ser Tyr Leu Ser Asn Val
835 840 845
Leu Asp Asp Asn Ile Cys Gly Ala Asp Lys Ala Pro Trp Thr Thr Tyr
850 855 860
Thr Thr Tyr Thr Thr Thr Glu Lys Cys Asn Lys Glu Arg Asp Lys Ser
865 870 875 880
Lys Ser Gln Ser Ser Asp Thr Leu Val Val Val Asn Val Pro Ser Pro
885 890 895
Leu Gly Asn Thr Pro Tyr Arg Tyr Lys Tyr Ala Cys Gln Cys Lys Ile
900 905 910
Pro Thr Asn Glu Glu Thr Cys Asp Asp Arg Lys Glu Tyr Met Asn Gln
915 920 925
Trp Ser Cys Gly Ser Ala Arg Thr Met Lys Arg Gly Tyr Lys Asn Asp
930 935 940
Asn Tyr Glu Leu Cys Lys Tyr Asn Gly Val Asp Val Lys Pro Thr Thr
945 950 955 960
Val Arg Ser Asn Ser Ser Lys Leu Asp Pro Glu Gly Gly Ser Leu Ala
965 970 975
Ala Leu Thr Ala His Gln Ala Cys His Leu Pro Leu Glu Thr Phe Thr
980 985 990
Arg His Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly Tyr
995 1000 1005
Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser
1010 1015 1020
Trp Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro
1025 1030 1035
Gly Ser Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu
1040 1045 1050
Gln Ala Arg Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg
1055 1060 1065
Phe Val Arg Gln Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Asn
1070 1075 1080
Gly Pro Ala Asp Ser Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro
1085 1090 1095
Thr Gly Ala Glu Phe Leu Gly Asp Gly Gly Asp Ile Ser Phe Ser
1100 1105 1110
Thr Arg Gly Thr Gln Asn Trp Thr Val Glu Arg Leu Leu Gln Ala
1115 1120 1125
His Arg Gln Leu Glu Glu Arg Gly Tyr Val Phe Val Gly Tyr His
1130 1135 1140
Gly Thr Phe Leu Glu Ala Ala Gln Ser Ile Val Phe Gly Gly Val
1145 1150 1155
Arg Ala Arg Ser Gln Asp Leu Asp Ala Ile Trp Arg Gly Phe Tyr
1160 1165 1170
Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly Tyr Ala Gln Asp Gln
1175 1180 1185
Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly Ala Leu Leu Arg
1190 1195 1200
Val Tyr Val Pro Arg Ser Ser Leu Pro Gly Phe Tyr Arg Thr Ser
1205 1210 1215
Leu Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu Val Glu Arg Leu
1220 1225 1230
Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala Ile Thr Gly Pro
1235 1240 1245
Glu Glu Glu Gly Gly Arg Leu Glu Thr Ile Leu Gly Trp Pro Leu
1250 1255 1260
Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr Asp Pro
1265 1270 1275
Arg Asn Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys
1280 1285 1290
Glu Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly
1295 1300 1305
Lys Pro Pro Arg Lys Asp Glu Leu
1310 1315
<210> 68
<211> 964
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 68
Leu Ser Phe Ile Leu Asn Ser Ser Asp Ala Asn Asn Pro Ser Glu Lys
1 5 10 15
Ile Gln Lys Asn Asn Asp Glu Val Cys Asn Cys Asn Glu Ser Gly Ile
20 25 30
Ala Ser Val Glu Gln Glu Gln Ile Ser Asp Pro Ser Ser Asn Lys Thr
35 40 45
Cys Ile Thr His Ser Ser Ile Lys Ala Asn Lys Lys Lys Val Cys Lys
50 55 60
His Val Lys Leu Gly Val Arg Glu Asn Asp Lys Asp Leu Arg Val Cys
65 70 75 80
Val Ile Glu His Thr Ser Leu Ser Gly Val Glu Asn Cys Cys Cys Gln
85 90 95
Asp Phe Leu Arg Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Ser Gly
100 105 110
Ser Ser Ser Asn Gly Ser Cys Asn Asn Lys Asn Gln Glu Ala Cys Glu
115 120 125
Lys Asn Leu Glu Lys Val Leu Ala Ser Leu Thr Asn Cys Tyr Lys Cys
130 135 140
Asp Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Lys Asn Trp Ile
145 150 155 160
Trp Lys Lys Ser Ser Gly Lys Glu Gly Gly Leu Gln Lys Glu Tyr Ala
165 170 175
Asn Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Cys Leu Val Val
180 185 190
Cys Leu Asp Glu Lys Gly Lys Lys Thr Gln Glu Leu Lys Asn Ile Arg
195 200 205
Thr Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu
210 215 220
Gly Lys Asn Leu Lys Pro Ser His Glu Lys Lys Asn Asp Asp Asn Gly
225 230 235 240
Lys Lys Leu Cys Lys Ala Leu Glu Tyr Ser Phe Ala Asp Tyr Gly Asp
245 250 255
Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp Leu
260 265 270
Glu Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe Arg Lys Tyr Ile
275 280 285
Lys Lys Asn Asn Thr Ala Glu Gln Asp Thr Ser Tyr Ser Ser Leu Asp
290 295 300
Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Leu
305 310 315 320
Ala Met Lys His Gly Ala Gly Met Asn Ser Thr Thr Cys Cys Gly Asp
325 330 335
Gly Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp Ile Pro Thr Ile
340 345 350
Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His
355 360 365
Phe Cys Lys Gln Arg Gln Glu Lys Val Lys Pro Val Ile Glu Asn Cys
370 375 380
Lys Ser Cys Lys Glu Ser Gly Gly Thr Cys Asn Gly Glu Cys Lys Thr
385 390 395 400
Glu Cys Lys Asn Lys Cys Glu Val Tyr Lys Lys Phe Ile Glu Asp Cys
405 410 415
Lys Gly Gly Asp Gly Thr Ala Gly Ser Ser Trp Val Lys Arg Trp Asp
420 425 430
Gln Ile Tyr Lys Arg Tyr Ser Lys Tyr Ile Glu Asp Ala Lys Arg Asn
435 440 445
Arg Lys Ala Gly Thr Lys Asn Cys Gly Pro Ser Ser Thr Thr Asn Ala
450 455 460
Ala Glu Asn Lys Cys Val Gln Ser Asp Ile Asp Ser Phe Phe Lys His
465 470 475 480
Leu Ile Asp Ile Gly Leu Thr Thr Pro Ser Ser Tyr Leu Ser Ile Val
485 490 495
Leu Asp Asp Asn Ile Cys Gly Ala Asp Lys Ala Pro Trp Thr Thr Tyr
500 505 510
Thr Thr Tyr Thr Thr Thr Glu Lys Cys Asn Lys Glu Thr Asp Lys Ser
515 520 525
Lys Leu Gln Gln Cys Asn Thr Ala Val Val Val Asn Val Pro Ser Pro
530 535 540
Leu Gly Asn Thr Pro His Gly Tyr Lys Tyr Ala Cys Gln Cys Lys Ile
545 550 555 560
Pro Thr Asn Glu Glu Thr Cys Asp Asp Arg Lys Glu Tyr Met Asn Gln
565 570 575
Trp Ser Cys Gly Ser Ala Arg Thr Met Lys Arg Gly Tyr Lys Asn Asp
580 585 590
Asn Tyr Glu Leu Cys Lys Tyr Asn Gly Val Asp Val Lys Pro Thr Thr
595 600 605
Val Arg Ser Asn Ser Ser Lys Leu Asp Pro Glu Gly Gly Ser Leu Ala
610 615 620
Ala Leu Thr Ala His Gln Ala Cys His Leu Pro Leu Glu Thr Phe Thr
625 630 635 640
Arg His Arg Gln Pro Arg Gly Trp Glu Gln Leu Glu Gln Cys Gly Tyr
645 650 655
Pro Val Gln Arg Leu Val Ala Leu Tyr Leu Ala Ala Arg Leu Ser Trp
660 665 670
Asn Gln Val Asp Gln Val Ile Arg Asn Ala Leu Ala Ser Pro Gly Ser
675 680 685
Gly Gly Asp Leu Gly Glu Ala Ile Arg Glu Gln Pro Glu Gln Ala Arg
690 695 700
Leu Ala Leu Thr Leu Ala Ala Ala Glu Ser Glu Arg Phe Val Arg Gln
705 710 715 720
Gly Thr Gly Asn Asp Glu Ala Gly Ala Ala Asn Gly Pro Ala Asp Ser
725 730 735
Gly Asp Ala Leu Leu Glu Arg Asn Tyr Pro Thr Gly Ala Glu Phe Leu
740 745 750
Gly Asp Gly Gly Asp Ile Ser Phe Ser Thr Arg Gly Thr Gln Asn Trp
755 760 765
Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg Gly
770 775 780
Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln Ser
785 790 795 800
Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala Ile
805 810 815
Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly Tyr
820 825 830
Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly Ala
835 840 845
Leu Leu Arg Val Tyr Val Pro Arg Ser Ser Leu Pro Gly Phe Tyr Arg
850 855 860
Thr Ser Leu Thr Leu Ala Ala Pro Glu Ala Ala Gly Glu Val Glu Arg
865 870 875 880
Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala Ile Thr Gly Pro
885 890 895
Glu Glu Glu Gly Gly Arg Leu Glu Thr Ile Leu Gly Trp Pro Leu Ala
900 905 910
Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr Asp Pro Arg Asn
915 920 925
Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp Lys Glu Gln Ala
930 935 940
Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly Lys Pro Pro Arg
945 950 955 960
Lys Asp Glu Leu
<210> 69
<211> 848
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 69
Leu Ser Phe Ile Leu Asn Ser Ser Asp Ala Asn Asn Pro Ser Glu Lys
1 5 10 15
Ile Gln Lys Asn Asn Asp Glu Val Cys Asn Cys Asn Glu Ser Gly Ile
20 25 30
Ala Ser Val Glu Gln Glu Gln Ile Ser Asp Pro Ser Ser Asn Lys Thr
35 40 45
Cys Ile Thr His Ser Ser Ile Lys Ala Asn Lys Lys Lys Val Cys Lys
50 55 60
His Val Lys Leu Gly Val Arg Glu Asn Asp Lys Asp Leu Arg Val Cys
65 70 75 80
Val Ile Glu His Thr Ser Leu Ser Gly Val Glu Asn Cys Cys Cys Gln
85 90 95
Asp Phe Leu Arg Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Ser Gly
100 105 110
Ser Ser Ser Asn Gly Ser Cys Asn Asn Lys Asn Gln Glu Ala Cys Glu
115 120 125
Lys Asn Leu Glu Lys Val Leu Ala Ser Leu Thr Asn Cys Tyr Lys Cys
130 135 140
Asp Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Lys Asn Trp Ile
145 150 155 160
Trp Lys Lys Ser Ser Gly Lys Glu Gly Gly Leu Gln Lys Glu Tyr Ala
165 170 175
Asn Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Cys Leu Val Val
180 185 190
Cys Leu Asp Glu Lys Gly Lys Lys Thr Gln Glu Leu Lys Asn Ile Arg
195 200 205
Thr Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu
210 215 220
Gly Lys Asn Leu Lys Pro Ser His Glu Lys Lys Asn Asp Asp Asn Gly
225 230 235 240
Lys Lys Leu Cys Lys Ala Leu Glu Tyr Ser Phe Ala Asp Tyr Gly Asp
245 250 255
Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp Leu
260 265 270
Glu Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe Arg Lys Tyr Ile
275 280 285
Lys Lys Asn Asn Thr Ala Glu Gln Asp Thr Ser Tyr Ser Ser Leu Asp
290 295 300
Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Leu
305 310 315 320
Ala Met Lys His Gly Ala Gly Met Asn Ser Thr Thr Cys Cys Gly Asp
325 330 335
Gly Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp Ile Pro Thr Ile
340 345 350
Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu His
355 360 365
Phe Cys Lys Gln Arg Gln Glu Lys Val Lys Pro Val Ile Glu Asn Cys
370 375 380
Lys Ser Cys Lys Glu Ser Gly Gly Thr Cys Asn Gly Glu Cys Lys Thr
385 390 395 400
Glu Cys Lys Asn Lys Cys Glu Val Tyr Lys Lys Phe Ile Glu Asp Cys
405 410 415
Lys Gly Gly Asp Gly Thr Ala Gly Ser Ser Trp Val Lys Arg Trp Asp
420 425 430
Gln Ile Tyr Lys Arg Tyr Ser Lys Tyr Ile Glu Asp Ala Lys Arg Asn
435 440 445
Arg Lys Ala Gly Thr Lys Asn Cys Gly Pro Ser Ser Thr Thr Asn Ala
450 455 460
Ala Glu Asn Lys Cys Val Gln Ser Asp Ile Asp Ser Phe Phe Lys His
465 470 475 480
Leu Ile Asp Ile Gly Leu Thr Thr Pro Ser Ser Tyr Leu Ser Ile Val
485 490 495
Leu Asp Asp Asn Ile Cys Gly Ala Asp Lys Ala Pro Trp Thr Thr Tyr
500 505 510
Thr Thr Tyr Thr Thr Thr Glu Lys Cys Asn Lys Glu Thr Asp Lys Ser
515 520 525
Lys Leu Gln Gln Cys Asn Thr Ala Val Val Val Asn Val Pro Ser Pro
530 535 540
Leu Gly Asn Thr Pro His Gly Tyr Lys Tyr Ala Cys Gln Cys Lys Ile
545 550 555 560
Pro Thr Asn Glu Glu Thr Cys Asp Asp Arg Lys Glu Tyr Met Asn Gln
565 570 575
Trp Ser Cys Gly Ser Ala Arg Thr Met Lys Arg Gly Tyr Lys Asn Asp
580 585 590
Asn Tyr Glu Leu Cys Lys Tyr Asn Gly Val Asp Val Lys Pro Thr Thr
595 600 605
Val Arg Ser Asn Ser Ser Lys Leu Asp Arg His Arg Gln Pro Arg Gly
610 615 620
Trp Glu Gln Leu Tyr Pro Thr Gly Ala Glu Phe Leu Gly Asp Gly Gly
625 630 635 640
Asp Ile Ser Phe Ser Thr Arg Gly Thr Gln Asn Trp Thr Val Glu Arg
645 650 655
Leu Leu Gln Ala His Arg Gln Leu Glu Glu Arg Gly Tyr Val Phe Val
660 665 670
Gly Tyr His Gly Thr Phe Leu Glu Ala Ala Gln Ser Ile Val Phe Gly
675 680 685
Gly Val Arg Ala Arg Ser Gln Asp Leu Asp Ala Ile Trp Arg Gly Phe
690 695 700
Tyr Ile Ala Gly Asp Pro Ala Leu Ala Tyr Gly Tyr Ala Gln Asp Gln
705 710 715 720
Glu Pro Asp Ala Arg Gly Arg Ile Arg Asn Gly Ala Leu Leu Arg Val
725 730 735
Tyr Val Pro Arg Ser Ser Leu Pro Gly Phe Tyr Arg Thr Ser Leu Thr
740 745 750
Leu Ala Ala Pro Glu Ala Ala Gly Glu Val Glu Arg Leu Ile Gly His
755 760 765
Pro Leu Pro Leu Arg Leu Asp Ala Ile Thr Gly Pro Glu Glu Glu Gly
770 775 780
Gly Arg Leu Glu Thr Ile Leu Gly Trp Pro Leu Ala Glu Arg Thr Val
785 790 795 800
Val Ile Pro Ser Ala Ile Pro Thr Asp Pro Arg Asn Val Gly Gly Asp
805 810 815
Leu Asp Pro Ser Ser Ile Pro Asp Lys Glu Gln Ala Ile Ser Ala Leu
820 825 830
Pro Asp Tyr Ala Ser Gln Pro Gly Lys Pro Pro Arg Lys Asp Glu Leu
835 840 845
<210> 70
<211> 808
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 70
Asn Tyr Ile Lys Gly Asp Pro Tyr Phe Ala Glu Tyr Ala Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Pro Ser Asp Ala Asn Asn Pro Ser Gly Glu Thr
20 25 30
Ala Asn His Asn Asp Glu Ala Cys Asn Cys Asn Glu Ser Gly Ile Ser
35 40 45
Ser Val Gly Gln Ala Gln Thr Ser Gly Pro Ser Ser Asn Lys Thr Cys
50 55 60
Ile Thr His Ser Ser Ile Lys Thr Asn Lys Lys Lys Glu Cys Lys Asp
65 70 75 80
Val Lys Leu Gly Val Arg Glu Asn Asp Lys Asp Leu Lys Ile Cys Val
85 90 95
Ile Glu Asp Thr Ser Leu Ser Gly Val Asp Asn Cys Cys Cys Gln Asp
100 105 110
Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Arg Gly Ser
115 120 125
Ser Ser Asn Asp Ser Cys Asp Asn Lys Asn Gln Asp Glu Cys Gln Lys
130 135 140
Lys Leu Glu Lys Val Phe Ala Ser Leu Thr Asn Gly Tyr Lys Cys Asp
145 150 155 160
Lys Cys Lys Ser Gly Thr Ser Arg Ser Lys Lys Lys Trp Ile Trp Lys
165 170 175
Lys Ser Ser Gly Asn Glu Glu Gly Leu Gln Glu Glu Tyr Ala Asn Thr
180 185 190
Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu Gly Asn Leu Pro
195 200 205
Lys Leu Glu Asn Val Cys Glu Asp Val Lys Asp Ile Asn Phe Asp Thr
210 215 220
Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Val Ser Phe His Glu Gly
225 230 235 240
Lys Asn Leu Lys Lys Arg Tyr Pro Gln Asn Lys Asn Ser Gly Asn Lys
245 250 255
Glu Asn Leu Cys Lys Ala Leu Glu Tyr Ser Phe Ala Asp Tyr Gly Asp
260 265 270
Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp Leu
275 280 285
Glu Leu Asn Leu Gln Asn Asn Phe Gly Lys Leu Phe Gly Lys Tyr Ile
290 295 300
Lys Lys Asn Asn Thr Ala Glu Gln Asp Thr Ser Tyr Ser Ser Leu Asp
305 310 315 320
Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Thr
325 330 335
Ala Met Lys His Gly Ala Glu Met Asn Ile Thr Thr Cys Asn Ala Asp
340 345 350
Gly Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp Ile Pro Thr Ile
355 360 365
Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu Asn
370 375 380
Phe Cys Glu Gln Arg Gln Ala Lys Val Lys Asp Val Ile Thr Asn Cys
385 390 395 400
Lys Ser Cys Lys Glu Ser Gly Asn Lys Cys Lys Thr Glu Cys Lys Thr
405 410 415
Lys Cys Lys Asp Glu Cys Glu Lys Tyr Lys Lys Phe Ile Glu Ala Cys
420 425 430
Gly Thr Ala Gly Gly Gly Ile Gly Thr Ala Gly Ser Pro Trp Ser Lys
435 440 445
Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys His Ile Glu Asp Ala
450 455 460
Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Thr Ser Ser Thr
465 470 475 480
Thr Asn Ala Ala Ala Ser Thr Asp Glu Asn Lys Cys Val Gln Ser Asp
485 490 495
Ile Asp Ser Phe Phe Lys His Leu Ile Asp Ile Gly Leu Thr Thr Pro
500 505 510
Ser Ser Tyr Leu Ser Asn Val Leu Asp Asp Asn Ile Cys Gly Ala Asp
515 520 525
Lys Ala Pro Trp Thr Thr Tyr Thr Thr Tyr Thr Thr Thr Glu Lys Cys
530 535 540
Asn Lys Glu Arg Asp Lys Ser Lys Ser Gln Ser Ser Asp Thr Leu Val
545 550 555 560
Val Val Asn Val Pro Ser Pro Leu Gly Asn Thr Pro Tyr Arg Tyr Lys
565 570 575
Tyr Arg His Arg Gln Pro Arg Gly Trp Glu Gln Leu Tyr Pro Thr Gly
580 585 590
Ala Glu Phe Leu Gly Asp Gly Gly Asp Ile Ser Phe Ser Thr Arg Gly
595 600 605
Thr Gln Asn Trp Thr Val Glu Arg Leu Leu Gln Ala His Arg Gln Leu
610 615 620
Glu Glu Arg Gly Tyr Val Phe Val Gly Tyr His Gly Thr Phe Leu Glu
625 630 635 640
Ala Ala Gln Ser Ile Val Phe Gly Gly Val Arg Ala Arg Ser Gln Asp
645 650 655
Leu Asp Ala Ile Trp Arg Gly Phe Tyr Ile Ala Gly Asp Pro Ala Leu
660 665 670
Ala Tyr Gly Tyr Ala Gln Asp Gln Glu Pro Asp Ala Arg Gly Arg Ile
675 680 685
Arg Asn Gly Ala Leu Leu Arg Val Tyr Val Pro Arg Ser Ser Leu Pro
690 695 700
Gly Phe Tyr Arg Thr Ser Leu Thr Leu Ala Ala Pro Glu Ala Ala Gly
705 710 715 720
Glu Val Glu Arg Leu Ile Gly His Pro Leu Pro Leu Arg Leu Asp Ala
725 730 735
Ile Thr Gly Pro Glu Glu Glu Gly Gly Arg Leu Glu Thr Ile Leu Gly
740 745 750
Trp Pro Leu Ala Glu Arg Thr Val Val Ile Pro Ser Ala Ile Pro Thr
755 760 765
Asp Pro Arg Asn Val Gly Gly Asp Leu Asp Pro Ser Ser Ile Pro Asp
770 775 780
Lys Glu Gln Ala Ile Ser Ala Leu Pro Asp Tyr Ala Ser Gln Pro Gly
785 790 795 800
Lys Pro Pro Arg Lys Asp Glu Leu
805
<210> 71
<211> 393
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 71
Met Gly Ala Asp Asp Val Val Asp Ser Ser Lys Ser Phe Val Met Glu
1 5 10 15
Asn Phe Ser Ser Tyr His Gly Thr Lys Pro Gly Tyr Val Asp Ser Ile
20 25 30
Gln Lys Gly Ile Gln Lys Pro Lys Ser Gly Thr Gln Gly Asn Tyr Asp
35 40 45
Asp Asp Trp Lys Gly Phe Tyr Ser Thr Asp Asn Lys Tyr Asp Ala Ala
50 55 60
Gly Tyr Ser Val Asp Asn Glu Asn Pro Leu Ser Gly Lys Ala Gly Gly
65 70 75 80
Val Val Lys Val Thr Tyr Pro Gly Leu Thr Lys Val Leu Ala Leu Lys
85 90 95
Val Asp Asn Ala Glu Thr Ile Lys Lys Glu Leu Gly Leu Ser Leu Thr
100 105 110
Glu Pro Leu Met Glu Gln Val Gly Thr Glu Glu Phe Ile Lys Arg Phe
115 120 125
Gly Asp Gly Ala Ser Arg Val Val Leu Ser Leu Pro Phe Ala Glu Gly
130 135 140
Ser Ser Ser Val Glu Tyr Ile Asn Asn Trp Glu Gln Ala Lys Ala Leu
145 150 155 160
Ser Val Glu Leu Glu Ile Asn Phe Glu Thr Arg Gly Lys Arg Gly Gln
165 170 175
Asp Ala Met Tyr Glu Tyr Met Ala Gln Ala Cys Ala Gly Asn Arg Val
180 185 190
Arg Arg Ser Val Gly Ser Ser Leu Ser Cys Ile Asn Leu Asp Trp Asp
195 200 205
Val Ile Arg Asp Lys Thr Lys Thr Lys Ile Glu Ser Leu Lys Glu His
210 215 220
Gly Pro Ile Lys Asn Lys Met Ser Glu Ser Pro Asn Lys Thr Val Ser
225 230 235 240
Glu Glu Lys Ala Lys Gln Tyr Leu Glu Glu Phe His Gln Thr Ala Leu
245 250 255
Glu His Pro Glu Leu Ser Glu Leu Lys Thr Val Thr Gly Thr Asn Pro
260 265 270
Val Phe Ala Gly Ala Asn Tyr Ala Ala Trp Ala Val Asn Val Ala Gln
275 280 285
Val Ile Asp Ser Glu Thr Ala Asp Asn Leu Glu Lys Thr Thr Ala Ala
290 295 300
Leu Ser Ile Leu Pro Gly Ile Gly Ser Val Met Gly Ile Ala Asp Gly
305 310 315 320
Ala Val His His Asn Thr Glu Glu Ile Val Ala Gln Ser Ile Ala Leu
325 330 335
Ser Ser Leu Met Val Ala Gln Ala Ile Pro Leu Val Gly Glu Leu Val
340 345 350
Asp Ile Gly Phe Ala Ala Tyr Asn Phe Val Glu Ser Ile Ile Asn Leu
355 360 365
Phe Gln Val Val His Asn Ser Tyr Asn Arg Pro Ala Tyr Ser Pro Gly
370 375 380
His Lys Thr Gln Pro Met His Glu Phe
385 390
<210> 72
<211> 1356
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 72
Met Gly Ala Asp Asp Val Val Asp Ser Ser Lys Ser Phe Val Met Glu
1 5 10 15
Asn Phe Ser Ser Tyr His Gly Thr Lys Pro Gly Tyr Val Asp Ser Ile
20 25 30
Gln Lys Gly Ile Gln Lys Pro Lys Ser Gly Thr Gln Gly Asn Tyr Asp
35 40 45
Asp Asp Trp Lys Gly Phe Tyr Ser Thr Asp Asn Lys Tyr Asp Ala Ala
50 55 60
Gly Tyr Ser Val Asp Asn Glu Asn Pro Leu Ser Gly Lys Ala Gly Gly
65 70 75 80
Val Val Lys Val Thr Tyr Pro Gly Leu Thr Lys Val Leu Ala Leu Lys
85 90 95
Val Asp Asn Ala Glu Thr Ile Lys Lys Glu Leu Gly Leu Ser Leu Thr
100 105 110
Glu Pro Leu Met Glu Gln Val Gly Thr Glu Glu Phe Ile Lys Arg Phe
115 120 125
Gly Asp Gly Ala Ser Arg Val Val Leu Ser Leu Pro Phe Ala Glu Gly
130 135 140
Ser Ser Ser Val Glu Tyr Ile Asn Asn Trp Glu Gln Ala Lys Ala Leu
145 150 155 160
Ser Val Glu Leu Glu Ile Asn Phe Glu Thr Arg Gly Lys Arg Gly Gln
165 170 175
Asp Ala Met Tyr Glu Tyr Met Ala Gln Ala Cys Ala Gly Asn Arg Val
180 185 190
Arg Arg Ser Val Gly Ser Ser Leu Ser Cys Ile Asn Leu Asp Trp Asp
195 200 205
Val Ile Arg Asp Lys Thr Lys Thr Lys Ile Glu Ser Leu Lys Glu His
210 215 220
Gly Pro Ile Lys Asn Lys Met Ser Glu Ser Pro Asn Lys Thr Val Ser
225 230 235 240
Glu Glu Lys Ala Lys Gln Tyr Leu Glu Glu Phe His Gln Thr Ala Leu
245 250 255
Glu His Pro Glu Leu Ser Glu Leu Lys Thr Val Thr Gly Thr Asn Pro
260 265 270
Val Phe Ala Gly Ala Asn Tyr Ala Ala Trp Ala Val Asn Val Ala Gln
275 280 285
Val Ile Asp Ser Glu Thr Ala Asp Asn Leu Glu Lys Thr Thr Ala Ala
290 295 300
Leu Ser Ile Leu Pro Gly Ile Gly Ser Val Met Gly Ile Ala Asp Gly
305 310 315 320
Ala Val His His Asn Thr Glu Glu Ile Val Ala Gln Ser Ile Ala Leu
325 330 335
Ser Ser Leu Met Val Ala Gln Ala Ile Pro Leu Val Gly Glu Leu Val
340 345 350
Asp Ile Gly Phe Ala Ala Tyr Asn Phe Val Glu Ser Ile Ile Asn Leu
355 360 365
Phe Gln Val Val His Asn Ser Tyr Asn Arg Pro Ala Tyr Ser Pro Gly
370 375 380
His Lys Thr Gln Pro Met His Glu Phe His Ser Asp Ser Gly Thr Asn
385 390 395 400
Asp Pro Cys Asp Arg Ile Pro Pro Pro Tyr Gly Asp Asn Asp Gln Trp
405 410 415
Lys Cys Ala Ile Ile Leu Ser Lys Val Ser Glu Lys Pro Glu Asn Val
420 425 430
Phe Val Pro Pro Arg Arg Gln Arg Met Cys Ile Asn Asn Leu Glu Lys
435 440 445
Leu Asn Val Asp Lys Ile Arg Asp Lys His Ala Phe Leu Ala Asp Val
450 455 460
Leu Leu Thr Ala Arg Asn Glu Gly Glu Arg Ile Val Gln Asn His Pro
465 470 475 480
Asp Thr Asn Ser Ser Asn Val Cys Asn Ala Leu Glu Arg Ser Phe Ala
485 490 495
Asp Ile Ala Asp Ile Ile Arg Gly Thr Asp Leu Trp Lys Gly Thr Asn
500 505 510
Ser Asn Leu Glu Gln Asn Leu Lys Gln Met Phe Ala Lys Ile Arg Glu
515 520 525
Asn Asp Lys Val Leu Gln Asp Lys Tyr Pro Lys Asp Gln Asn Tyr Arg
530 535 540
Lys Leu Arg Glu Asp Trp Trp Asn Ala Asn Arg Gln Lys Val Trp Glu
545 550 555 560
Val Ile Thr Cys Gly Ala Arg Ser Asn Asp Leu Leu Ile Lys Arg Gly
565 570 575
Trp Arg Thr Ser Gly Lys Ser Asn Gly Asp Asn Lys Leu Glu Leu Cys
580 585 590
Arg Lys Cys Gly His Tyr Glu Glu Lys Val Pro Thr Lys Leu Asp Tyr
595 600 605
Val Pro Gln Phe Leu Arg Trp Leu Thr Glu Trp Ile Glu Asp Phe Tyr
610 615 620
Arg Glu Lys Gln Asn Leu Ile Asp Asp Met Glu Arg His Arg Glu Glu
625 630 635 640
Cys Thr Ser Glu Asp His Lys Ser Lys Glu Gly Thr Ser Tyr Cys Ser
645 650 655
Thr Cys Lys Asp Lys Cys Lys Lys Tyr Cys Glu Cys Val Lys Lys Trp
660 665 670
Lys Ser Glu Trp Glu Asn Gln Lys Asn Lys Tyr Thr Glu Leu Tyr Gln
675 680 685
Gln Asn Lys Asn Glu Thr Ser Gln Lys Asn Thr Ser Arg Tyr Asp Asp
690 695 700
Tyr Val Lys Asp Phe Phe Lys Lys Leu Glu Ala Asn Tyr Ser Ser Leu
705 710 715 720
Glu Asn Tyr Ile Lys Gly Asp Pro Tyr Phe Ala Glu Tyr Ala Thr Lys
725 730 735
Leu Ser Phe Ile Leu Asn Ser Ser Asp Ala Asn Asn Pro Ser Glu Lys
740 745 750
Ile Gln Lys Asn Asn Asp Glu Val Cys Asn Cys Asn Glu Ser Gly Ile
755 760 765
Ala Ser Val Glu Gln Glu Gln Ile Ser Asp Pro Ser Ser Asn Lys Thr
770 775 780
Cys Ile Thr His Ser Ser Ile Lys Ala Asn Lys Lys Lys Val Cys Lys
785 790 795 800
His Val Lys Leu Gly Val Arg Glu Asn Asp Lys Asp Leu Arg Val Cys
805 810 815
Val Ile Glu His Thr Ser Leu Ser Gly Val Glu Asn Cys Cys Cys Gln
820 825 830
Asp Phe Leu Arg Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Ser Gly
835 840 845
Ser Ser Ser Asn Gly Ser Cys Asn Asn Lys Asn Gln Glu Ala Cys Glu
850 855 860
Lys Asn Leu Glu Lys Val Leu Ala Ser Leu Thr Asn Cys Tyr Lys Cys
865 870 875 880
Asp Lys Cys Lys Ser Glu Gln Ser Lys Lys Asn Asn Lys Asn Trp Ile
885 890 895
Trp Lys Lys Ser Ser Gly Lys Glu Gly Gly Leu Gln Lys Glu Tyr Ala
900 905 910
Asn Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Cys Leu Val Val
915 920 925
Cys Leu Asp Glu Lys Gly Lys Lys Thr Gln Glu Leu Lys Asn Ile Arg
930 935 940
Thr Asn Ser Glu Leu Leu Lys Glu Trp Ile Ile Ala Ala Phe His Glu
945 950 955 960
Gly Lys Asn Leu Lys Pro Ser His Glu Lys Lys Asn Asp Asp Asn Gly
965 970 975
Lys Lys Leu Cys Lys Ala Leu Glu Tyr Ser Phe Ala Asp Tyr Gly Asp
980 985 990
Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp Leu
995 1000 1005
Glu Leu Asn Leu Gln Lys Ile Phe Gly Lys Leu Phe Arg Lys Tyr
1010 1015 1020
Ile Lys Lys Asn Asn Thr Ala Glu Gln Asp Thr Ser Tyr Ser Ser
1025 1030 1035
Leu Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr
1040 1045 1050
Ile Trp Leu Ala Met Lys His Gly Ala Gly Met Asn Ser Thr Thr
1055 1060 1065
Cys Cys Gly Asp Gly Ser Val Thr Gly Ser Gly Ser Ser Cys Asp
1070 1075 1080
Asp Ile Pro Thr Ile Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu
1085 1090 1095
Gln Glu Trp Val Glu His Phe Cys Lys Gln Arg Gln Glu Lys Val
1100 1105 1110
Lys Pro Val Ile Glu Asn Cys Lys Ser Cys Lys Glu Ser Gly Gly
1115 1120 1125
Thr Cys Asn Gly Glu Cys Lys Thr Glu Cys Lys Asn Lys Cys Glu
1130 1135 1140
Val Tyr Lys Lys Phe Ile Glu Asp Cys Lys Gly Gly Asp Gly Thr
1145 1150 1155
Ala Gly Ser Ser Trp Val Lys Arg Trp Asp Gln Ile Tyr Lys Arg
1160 1165 1170
Tyr Ser Lys Tyr Ile Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly
1175 1180 1185
Thr Lys Asn Cys Gly Pro Ser Ser Thr Thr Asn Ala Ala Glu Asn
1190 1195 1200
Lys Cys Val Gln Ser Asp Ile Asp Ser Phe Phe Lys His Leu Ile
1205 1210 1215
Asp Ile Gly Leu Thr Thr Pro Ser Ser Tyr Leu Ser Ile Val Leu
1220 1225 1230
Asp Asp Asn Ile Cys Gly Ala Asp Lys Ala Pro Trp Thr Thr Tyr
1235 1240 1245
Thr Thr Tyr Thr Thr Thr Glu Lys Cys Asn Lys Glu Thr Asp Lys
1250 1255 1260
Ser Lys Leu Gln Gln Cys Asn Thr Ala Val Val Val Asn Val Pro
1265 1270 1275
Ser Pro Leu Gly Asn Thr Pro His Gly Tyr Lys Tyr Ala Cys Gln
1280 1285 1290
Cys Lys Ile Pro Thr Asn Glu Glu Thr Cys Asp Asp Arg Lys Glu
1295 1300 1305
Tyr Met Asn Gln Trp Ser Cys Gly Ser Ala Arg Thr Met Lys Arg
1310 1315 1320
Gly Tyr Lys Asn Asp Asn Tyr Glu Leu Cys Lys Tyr Asn Gly Val
1325 1330 1335
Asp Val Lys Pro Thr Thr Val Arg Ser Asn Ser Ser Lys Leu Asp
1340 1345 1350
Ser Gly Arg
1355
<210> 73
<211> 1364
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 73
Met Gly Ala Asp Asp Val Val Asp Ser Ser Lys Ser Phe Val Met Glu
1 5 10 15
Asn Phe Ser Ser Tyr His Gly Thr Lys Pro Gly Tyr Val Asp Ser Ile
20 25 30
Gln Lys Gly Ile Gln Lys Pro Lys Ser Gly Thr Gln Gly Asn Tyr Asp
35 40 45
Asp Asp Trp Lys Gly Phe Tyr Ser Thr Asp Asn Lys Tyr Asp Ala Ala
50 55 60
Gly Tyr Ser Val Asp Asn Glu Asn Pro Leu Ser Gly Lys Ala Gly Gly
65 70 75 80
Val Val Lys Val Thr Tyr Pro Gly Leu Thr Lys Val Leu Ala Leu Lys
85 90 95
Val Asp Asn Ala Glu Thr Ile Lys Lys Glu Leu Gly Leu Ser Leu Thr
100 105 110
Glu Pro Leu Met Glu Gln Val Gly Thr Glu Glu Phe Ile Lys Arg Phe
115 120 125
Gly Asp Gly Ala Ser Arg Val Val Leu Ser Leu Pro Phe Ala Glu Gly
130 135 140
Ser Ser Ser Val Glu Tyr Ile Asn Asn Trp Glu Gln Ala Lys Ala Leu
145 150 155 160
Ser Val Glu Leu Glu Ile Asn Phe Glu Thr Arg Gly Lys Arg Gly Gln
165 170 175
Asp Ala Met Tyr Glu Tyr Met Ala Gln Ala Cys Ala Gly Asn Arg Val
180 185 190
Arg Arg Ser Val Gly Ser Ser Leu Ser Cys Ile Asn Leu Asp Trp Asp
195 200 205
Val Ile Arg Asp Lys Thr Lys Thr Lys Ile Glu Ser Leu Lys Glu His
210 215 220
Gly Pro Ile Lys Asn Lys Met Ser Glu Ser Pro Asn Lys Thr Val Ser
225 230 235 240
Glu Glu Lys Ala Lys Gln Tyr Leu Glu Glu Phe His Gln Thr Ala Leu
245 250 255
Glu His Pro Glu Leu Ser Glu Leu Lys Thr Val Thr Gly Thr Asn Pro
260 265 270
Val Phe Ala Gly Ala Asn Tyr Ala Ala Trp Ala Val Asn Val Ala Gln
275 280 285
Val Ile Asp Ser Glu Thr Ala Asp Asn Leu Glu Lys Thr Thr Ala Ala
290 295 300
Leu Ser Ile Leu Pro Gly Ile Gly Ser Val Met Gly Ile Ala Asp Gly
305 310 315 320
Ala Val His His Asn Thr Glu Glu Ile Val Ala Gln Ser Ile Ala Leu
325 330 335
Ser Ser Leu Met Val Ala Gln Ala Ile Pro Leu Val Gly Glu Leu Val
340 345 350
Asp Ile Gly Phe Ala Ala Tyr Asn Phe Val Glu Ser Ile Ile Asn Leu
355 360 365
Phe Gln Val Val His Asn Ser Tyr Asn Arg Pro Ala Tyr Ser Pro Gly
370 375 380
His Lys Thr Gln Pro Met His Glu Phe His Ser Asp Ser Gly Lys Tyr
385 390 395 400
Asp Pro Cys Glu Lys Lys Leu Pro Pro Tyr Asp Asp Asn Asp Gln Trp
405 410 415
Lys Cys Gln Gln Asn Ser Ser Asp Gly Ser Gly Lys Pro Glu Asn Ile
420 425 430
Cys Val Pro Pro Arg Arg Glu Arg Leu Cys Thr Tyr Asn Leu Glu Asn
435 440 445
Leu Lys Phe Asp Lys Ile Arg Asp Asn Asn Ala Phe Leu Ala Asp Val
450 455 460
Leu Leu Thr Ala Arg Asn Glu Gly Glu Lys Ile Val Gln Asn His Pro
465 470 475 480
Asp Thr Asn Ser Ser Asn Val Cys Asn Ala Leu Glu Arg Ser Phe Ala
485 490 495
Asp Leu Ala Asp Ile Ile Arg Gly Thr Asp Gln Trp Lys Gly Thr Asn
500 505 510
Ser Asn Leu Glu Lys Asn Leu Lys Gln Met Phe Ala Lys Ile Arg Glu
515 520 525
Asn Asp Lys Val Leu Gln Asp Lys Tyr Pro Lys Asp Gln Lys Tyr Thr
530 535 540
Lys Leu Arg Glu Ala Trp Trp Asn Ala Asn Arg Gln Lys Val Trp Glu
545 550 555 560
Val Ile Thr Cys Gly Ala Arg Ser Asn Asp Leu Leu Ile Lys Arg Gly
565 570 575
Trp Arg Thr Ser Gly Lys Ser Asp Arg Lys Lys Asn Phe Glu Leu Cys
580 585 590
Arg Lys Cys Gly His Tyr Glu Lys Glu Val Pro Thr Lys Leu Asp Tyr
595 600 605
Val Pro Gln Phe Leu Arg Trp Leu Thr Glu Trp Ile Glu Asp Phe Tyr
610 615 620
Arg Glu Lys Gln Asn Leu Ile Asp Asp Met Glu Arg His Arg Glu Glu
625 630 635 640
Cys Thr Arg Glu Asp His Lys Ser Lys Glu Gly Thr Ser Tyr Cys Ser
645 650 655
Thr Cys Lys Asp Lys Cys Lys Lys Tyr Cys Glu Cys Val Lys Lys Trp
660 665 670
Lys Thr Glu Trp Glu Asn Gln Glu Asn Lys Tyr Lys Asp Leu Tyr Glu
675 680 685
Gln Asn Lys Asn Lys Thr Ser Gln Lys Asn Thr Ser Arg Tyr Asp Asp
690 695 700
Tyr Val Lys Asp Phe Phe Glu Lys Leu Glu Ala Asn Tyr Ser Ser Leu
705 710 715 720
Glu Asn Tyr Ile Lys Gly Asp Pro Tyr Phe Ala Glu Tyr Ala Thr Lys
725 730 735
Leu Ser Phe Ile Leu Asn Pro Ser Asp Ala Asn Asn Pro Ser Gly Glu
740 745 750
Thr Ala Asn His Asn Asp Glu Ala Cys Asn Cys Asn Glu Ser Gly Ile
755 760 765
Ser Ser Val Gly Gln Ala Gln Thr Ser Gly Pro Ser Ser Asn Lys Thr
770 775 780
Cys Ile Thr His Ser Ser Ile Lys Thr Asn Lys Lys Lys Glu Cys Lys
785 790 795 800
Asp Val Lys Leu Gly Val Arg Glu Asn Asp Lys Asp Leu Lys Ile Cys
805 810 815
Val Ile Glu Asp Thr Ser Leu Ser Gly Val Asp Asn Cys Cys Cys Gln
820 825 830
Asp Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Arg Gly
835 840 845
Ser Ser Ser Asn Asp Ser Cys Asp Asn Lys Asn Gln Asp Glu Cys Gln
850 855 860
Lys Lys Leu Glu Lys Val Phe Ala Ser Leu Thr Asn Gly Tyr Lys Cys
865 870 875 880
Asp Lys Cys Lys Ser Gly Thr Ser Arg Ser Lys Lys Lys Trp Ile Trp
885 890 895
Lys Lys Ser Ser Gly Asn Glu Glu Gly Leu Gln Glu Glu Tyr Ala Asn
900 905 910
Thr Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu Gly Asn Leu
915 920 925
Pro Lys Leu Glu Asn Val Cys Glu Asp Val Lys Asp Ile Asn Phe Asp
930 935 940
Thr Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Val Ser Phe His Glu
945 950 955 960
Gly Lys Asn Leu Lys Lys Arg Tyr Pro Gln Asn Lys Asn Ser Gly Asn
965 970 975
Lys Glu Asn Leu Cys Lys Ala Leu Glu Tyr Ser Phe Ala Asp Tyr Gly
980 985 990
Asp Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp
995 1000 1005
Leu Glu Leu Asn Leu Gln Asn Asn Phe Gly Lys Leu Phe Gly Lys
1010 1015 1020
Tyr Ile Lys Lys Asn Asn Thr Ala Glu Gln Asp Thr Ser Tyr Ser
1025 1030 1035
Ser Leu Asp Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys
1040 1045 1050
Tyr Ile Trp Thr Ala Met Lys His Gly Ala Glu Met Asn Ile Thr
1055 1060 1065
Thr Cys Asn Ala Asp Gly Ser Val Thr Gly Ser Gly Ser Ser Cys
1070 1075 1080
Asp Asp Ile Pro Thr Ile Asp Leu Ile Pro Gln Tyr Leu Arg Phe
1085 1090 1095
Leu Gln Glu Trp Val Glu Asn Phe Cys Glu Gln Arg Gln Ala Lys
1100 1105 1110
Val Lys Asp Val Ile Thr Asn Cys Lys Ser Cys Lys Glu Ser Gly
1115 1120 1125
Asn Lys Cys Lys Thr Glu Cys Lys Thr Lys Cys Lys Asp Glu Cys
1130 1135 1140
Glu Lys Tyr Lys Lys Phe Ile Glu Ala Cys Gly Thr Ala Gly Gly
1145 1150 1155
Gly Ile Gly Thr Ala Gly Ser Pro Trp Ser Lys Arg Trp Asp Gln
1160 1165 1170
Ile Tyr Lys Arg Tyr Ser Lys His Ile Glu Asp Ala Lys Arg Asn
1175 1180 1185
Arg Lys Ala Gly Thr Lys Asn Cys Gly Thr Ser Ser Thr Thr Asn
1190 1195 1200
Ala Ala Ala Ser Thr Asp Glu Asn Lys Cys Val Gln Ser Asp Ile
1205 1210 1215
Asp Ser Phe Phe Lys His Leu Ile Asp Ile Gly Leu Thr Thr Pro
1220 1225 1230
Ser Ser Tyr Leu Ser Asn Val Leu Asp Asp Asn Ile Cys Gly Ala
1235 1240 1245
Asp Lys Ala Pro Trp Thr Thr Tyr Thr Thr Tyr Thr Thr Thr Glu
1250 1255 1260
Lys Cys Asn Lys Glu Arg Asp Lys Ser Lys Ser Gln Ser Ser Asp
1265 1270 1275
Thr Leu Val Val Val Asn Val Pro Ser Pro Leu Gly Asn Thr Pro
1280 1285 1290
Tyr Arg Tyr Lys Tyr Ala Cys Gln Cys Lys Ile Pro Thr Asn Glu
1295 1300 1305
Glu Thr Cys Asp Asp Arg Lys Glu Tyr Met Asn Gln Trp Ser Cys
1310 1315 1320
Gly Ser Ala Arg Thr Met Lys Arg Gly Tyr Lys Asn Asp Asn Tyr
1325 1330 1335
Glu Leu Cys Lys Tyr Asn Gly Val Asp Val Lys Pro Thr Thr Val
1340 1345 1350
Arg Ser Asn Ser Ser Lys Leu Asp Ser Gly Arg
1355 1360
<210> 74
<211> 1013
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 74
Met Gly Ala Asp Asp Val Val Asp Ser Ser Lys Ser Phe Val Met Glu
1 5 10 15
Asn Phe Ser Ser Tyr His Gly Thr Lys Pro Gly Tyr Val Asp Ser Ile
20 25 30
Gln Lys Gly Ile Gln Lys Pro Lys Ser Gly Thr Gln Gly Asn Tyr Asp
35 40 45
Asp Asp Trp Lys Gly Phe Tyr Ser Thr Asp Asn Lys Tyr Asp Ala Ala
50 55 60
Gly Tyr Ser Val Asp Asn Glu Asn Pro Leu Ser Gly Lys Ala Gly Gly
65 70 75 80
Val Val Lys Val Thr Tyr Pro Gly Leu Thr Lys Val Leu Ala Leu Lys
85 90 95
Val Asp Asn Ala Glu Thr Ile Lys Lys Glu Leu Gly Leu Ser Leu Thr
100 105 110
Glu Pro Leu Met Glu Gln Val Gly Thr Glu Glu Phe Ile Lys Arg Phe
115 120 125
Gly Asp Gly Ala Ser Arg Val Val Leu Ser Leu Pro Phe Ala Glu Gly
130 135 140
Ser Ser Ser Val Glu Tyr Ile Asn Asn Trp Glu Gln Ala Lys Ala Leu
145 150 155 160
Ser Val Glu Leu Glu Ile Asn Phe Glu Thr Arg Gly Lys Arg Gly Gln
165 170 175
Asp Ala Met Tyr Glu Tyr Met Ala Gln Ala Cys Ala Gly Asn Arg Val
180 185 190
Arg Arg Ser Val Gly Ser Ser Leu Ser Cys Ile Asn Leu Asp Trp Asp
195 200 205
Val Ile Arg Asp Lys Thr Lys Thr Lys Ile Glu Ser Leu Lys Glu His
210 215 220
Gly Pro Ile Lys Asn Lys Met Ser Glu Ser Pro Asn Lys Thr Val Ser
225 230 235 240
Glu Glu Lys Ala Lys Gln Tyr Leu Glu Glu Phe His Gln Thr Ala Leu
245 250 255
Glu His Pro Glu Leu Ser Glu Leu Lys Thr Val Thr Gly Thr Asn Pro
260 265 270
Val Phe Ala Gly Ala Asn Tyr Ala Ala Trp Ala Val Asn Val Ala Gln
275 280 285
Val Ile Asp Ser Glu Thr Ala Asp Asn Leu Glu Lys Thr Thr Ala Ala
290 295 300
Leu Ser Ile Leu Pro Gly Ile Gly Ser Val Met Gly Ile Ala Asp Gly
305 310 315 320
Ala Val His His Asn Thr Glu Glu Ile Val Ala Gln Ser Ile Ala Leu
325 330 335
Ser Ser Leu Met Val Ala Gln Ala Ile Pro Leu Val Gly Glu Leu Val
340 345 350
Asp Ile Gly Phe Ala Ala Tyr Asn Phe Val Glu Ser Ile Ile Asn Leu
355 360 365
Phe Gln Val Val His Asn Ser Tyr Asn Arg Pro Ala Tyr Ser Pro Gly
370 375 380
His Lys Thr Gln Pro Met His Glu Phe Leu Ser Phe Ile Leu Asn Ser
385 390 395 400
Ser Asp Ala Asn Asn Pro Ser Glu Lys Ile Gln Lys Asn Asn Asp Glu
405 410 415
Val Cys Asn Cys Asn Glu Ser Gly Ile Ala Ser Val Glu Gln Glu Gln
420 425 430
Ile Ser Asp Pro Ser Ser Asn Lys Thr Cys Ile Thr His Ser Ser Ile
435 440 445
Lys Ala Asn Lys Lys Lys Val Cys Lys His Val Lys Leu Gly Val Arg
450 455 460
Glu Asn Asp Lys Asp Leu Arg Val Cys Val Ile Glu His Thr Ser Leu
465 470 475 480
Ser Gly Val Glu Asn Cys Cys Cys Gln Asp Phe Leu Arg Ile Leu Gln
485 490 495
Glu Asn Cys Ser Asp Asn Lys Ser Gly Ser Ser Ser Asn Gly Ser Cys
500 505 510
Asn Asn Lys Asn Gln Glu Ala Cys Glu Lys Asn Leu Glu Lys Val Leu
515 520 525
Ala Ser Leu Thr Asn Cys Tyr Lys Cys Asp Lys Cys Lys Ser Glu Gln
530 535 540
Ser Lys Lys Asn Asn Lys Asn Trp Ile Trp Lys Lys Ser Ser Gly Lys
545 550 555 560
Glu Gly Gly Leu Gln Lys Glu Tyr Ala Asn Thr Ile Gly Leu Pro Pro
565 570 575
Arg Thr Gln Ser Leu Cys Leu Val Val Cys Leu Asp Glu Lys Gly Lys
580 585 590
Lys Thr Gln Glu Leu Lys Asn Ile Arg Thr Asn Ser Glu Leu Leu Lys
595 600 605
Glu Trp Ile Ile Ala Ala Phe His Glu Gly Lys Asn Leu Lys Pro Ser
610 615 620
His Glu Lys Lys Asn Asp Asp Asn Gly Lys Lys Leu Cys Lys Ala Leu
625 630 635 640
Glu Tyr Ser Phe Ala Asp Tyr Gly Asp Leu Ile Lys Gly Thr Ser Ile
645 650 655
Trp Asp Asn Glu Tyr Thr Lys Asp Leu Glu Leu Asn Leu Gln Lys Ile
660 665 670
Phe Gly Lys Leu Phe Arg Lys Tyr Ile Lys Lys Asn Asn Thr Ala Glu
675 680 685
Gln Asp Thr Ser Tyr Ser Ser Leu Asp Glu Leu Arg Glu Ser Trp Trp
690 695 700
Asn Thr Asn Lys Lys Tyr Ile Trp Leu Ala Met Lys His Gly Ala Gly
705 710 715 720
Met Asn Ser Thr Thr Cys Cys Gly Asp Gly Ser Val Thr Gly Ser Gly
725 730 735
Ser Ser Cys Asp Asp Ile Pro Thr Ile Asp Leu Ile Pro Gln Tyr Leu
740 745 750
Arg Phe Leu Gln Glu Trp Val Glu His Phe Cys Lys Gln Arg Gln Glu
755 760 765
Lys Val Lys Pro Val Ile Glu Asn Cys Lys Ser Cys Lys Glu Ser Gly
770 775 780
Gly Thr Cys Asn Gly Glu Cys Lys Thr Glu Cys Lys Asn Lys Cys Glu
785 790 795 800
Val Tyr Lys Lys Phe Ile Glu Asp Cys Lys Gly Gly Asp Gly Thr Ala
805 810 815
Gly Ser Ser Trp Val Lys Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser
820 825 830
Lys Tyr Ile Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn
835 840 845
Cys Gly Pro Ser Ser Thr Thr Asn Ala Ala Glu Asn Lys Cys Val Gln
850 855 860
Ser Asp Ile Asp Ser Phe Phe Lys His Leu Ile Asp Ile Gly Leu Thr
865 870 875 880
Thr Pro Ser Ser Tyr Leu Ser Ile Val Leu Asp Asp Asn Ile Cys Gly
885 890 895
Ala Asp Lys Ala Pro Trp Thr Thr Tyr Thr Thr Tyr Thr Thr Thr Glu
900 905 910
Lys Cys Asn Lys Glu Thr Asp Lys Ser Lys Leu Gln Gln Cys Asn Thr
915 920 925
Ala Val Val Val Asn Val Pro Ser Pro Leu Gly Asn Thr Pro His Gly
930 935 940
Tyr Lys Tyr Ala Cys Gln Cys Lys Ile Pro Thr Asn Glu Glu Thr Cys
945 950 955 960
Asp Asp Arg Lys Glu Tyr Met Asn Gln Trp Ser Cys Gly Ser Ala Arg
965 970 975
Thr Met Lys Arg Gly Tyr Lys Asn Asp Asn Tyr Glu Leu Cys Lys Tyr
980 985 990
Asn Gly Val Asp Val Lys Pro Thr Thr Val Arg Ser Asn Ser Ser Lys
995 1000 1005
Leu Asp Ser Gly Arg
1010
<210> 75
<211> 1036
<212> PRT
<213> 人工
<220>
<223> 人工构建体
<400> 75
Met Gly Ala Asp Asp Val Val Asp Ser Ser Lys Ser Phe Val Met Glu
1 5 10 15
Asn Phe Ser Ser Tyr His Gly Thr Lys Pro Gly Tyr Val Asp Ser Ile
20 25 30
Gln Lys Gly Ile Gln Lys Pro Lys Ser Gly Thr Gln Gly Asn Tyr Asp
35 40 45
Asp Asp Trp Lys Gly Phe Tyr Ser Thr Asp Asn Lys Tyr Asp Ala Ala
50 55 60
Gly Tyr Ser Val Asp Asn Glu Asn Pro Leu Ser Gly Lys Ala Gly Gly
65 70 75 80
Val Val Lys Val Thr Tyr Pro Gly Leu Thr Lys Val Leu Ala Leu Lys
85 90 95
Val Asp Asn Ala Glu Thr Ile Lys Lys Glu Leu Gly Leu Ser Leu Thr
100 105 110
Glu Pro Leu Met Glu Gln Val Gly Thr Glu Glu Phe Ile Lys Arg Phe
115 120 125
Gly Asp Gly Ala Ser Arg Val Val Leu Ser Leu Pro Phe Ala Glu Gly
130 135 140
Ser Ser Ser Val Glu Tyr Ile Asn Asn Trp Glu Gln Ala Lys Ala Leu
145 150 155 160
Ser Val Glu Leu Glu Ile Asn Phe Glu Thr Arg Gly Lys Arg Gly Gln
165 170 175
Asp Ala Met Tyr Glu Tyr Met Ala Gln Ala Cys Ala Gly Asn Arg Val
180 185 190
Arg Arg Ser Val Gly Ser Ser Leu Ser Cys Ile Asn Leu Asp Trp Asp
195 200 205
Val Ile Arg Asp Lys Thr Lys Thr Lys Ile Glu Ser Leu Lys Glu His
210 215 220
Gly Pro Ile Lys Asn Lys Met Ser Glu Ser Pro Asn Lys Thr Val Ser
225 230 235 240
Glu Glu Lys Ala Lys Gln Tyr Leu Glu Glu Phe His Gln Thr Ala Leu
245 250 255
Glu His Pro Glu Leu Ser Glu Leu Lys Thr Val Thr Gly Thr Asn Pro
260 265 270
Val Phe Ala Gly Ala Asn Tyr Ala Ala Trp Ala Val Asn Val Ala Gln
275 280 285
Val Ile Asp Ser Glu Thr Ala Asp Asn Leu Glu Lys Thr Thr Ala Ala
290 295 300
Leu Ser Ile Leu Pro Gly Ile Gly Ser Val Met Gly Ile Ala Asp Gly
305 310 315 320
Ala Val His His Asn Thr Glu Glu Ile Val Ala Gln Ser Ile Ala Leu
325 330 335
Ser Ser Leu Met Val Ala Gln Ala Ile Pro Leu Val Gly Glu Leu Val
340 345 350
Asp Ile Gly Phe Ala Ala Tyr Asn Phe Val Glu Ser Ile Ile Asn Leu
355 360 365
Phe Gln Val Val His Asn Ser Tyr Asn Arg Pro Ala Tyr Ser Pro Gly
370 375 380
His Lys Thr Gln Pro Met His Glu Phe Asn Tyr Ile Lys Gly Asp Pro
385 390 395 400
Tyr Phe Ala Glu Tyr Ala Thr Lys Leu Ser Phe Ile Leu Asn Pro Ser
405 410 415
Asp Ala Asn Asn Pro Ser Gly Glu Thr Ala Asn His Asn Asp Glu Ala
420 425 430
Cys Asn Cys Asn Glu Ser Gly Ile Ser Ser Val Gly Gln Ala Gln Thr
435 440 445
Ser Gly Pro Ser Ser Asn Lys Thr Cys Ile Thr His Ser Ser Ile Lys
450 455 460
Thr Asn Lys Lys Lys Glu Cys Lys Asp Val Lys Leu Gly Val Arg Glu
465 470 475 480
Asn Asp Lys Asp Leu Lys Ile Cys Val Ile Glu Asp Thr Ser Leu Ser
485 490 495
Gly Val Asp Asn Cys Cys Cys Gln Asp Leu Leu Gly Ile Leu Gln Glu
500 505 510
Asn Cys Ser Asp Asn Lys Arg Gly Ser Ser Ser Asn Asp Ser Cys Asp
515 520 525
Asn Lys Asn Gln Asp Glu Cys Gln Lys Lys Leu Glu Lys Val Phe Ala
530 535 540
Ser Leu Thr Asn Gly Tyr Lys Cys Asp Lys Cys Lys Ser Gly Thr Ser
545 550 555 560
Arg Ser Lys Lys Lys Trp Ile Trp Lys Lys Ser Ser Gly Asn Glu Glu
565 570 575
Gly Leu Gln Glu Glu Tyr Ala Asn Thr Ile Gly Leu Pro Pro Arg Thr
580 585 590
Gln Ser Leu Tyr Leu Gly Asn Leu Pro Lys Leu Glu Asn Val Cys Glu
595 600 605
Asp Val Lys Asp Ile Asn Phe Asp Thr Lys Glu Lys Phe Leu Ala Gly
610 615 620
Cys Leu Ile Val Ser Phe His Glu Gly Lys Asn Leu Lys Lys Arg Tyr
625 630 635 640
Pro Gln Asn Lys Asn Ser Gly Asn Lys Glu Asn Leu Cys Lys Ala Leu
645 650 655
Glu Tyr Ser Phe Ala Asp Tyr Gly Asp Leu Ile Lys Gly Thr Ser Ile
660 665 670
Trp Asp Asn Glu Tyr Thr Lys Asp Leu Glu Leu Asn Leu Gln Asn Asn
675 680 685
Phe Gly Lys Leu Phe Gly Lys Tyr Ile Lys Lys Asn Asn Thr Ala Glu
690 695 700
Gln Asp Thr Ser Tyr Ser Ser Leu Asp Glu Leu Arg Glu Ser Trp Trp
705 710 715 720
Asn Thr Asn Lys Lys Tyr Ile Trp Thr Ala Met Lys His Gly Ala Glu
725 730 735
Met Asn Ile Thr Thr Cys Asn Ala Asp Gly Ser Val Thr Gly Ser Gly
740 745 750
Ser Ser Cys Asp Asp Ile Pro Thr Ile Asp Leu Ile Pro Gln Tyr Leu
755 760 765
Arg Phe Leu Gln Glu Trp Val Glu Asn Phe Cys Glu Gln Arg Gln Ala
770 775 780
Lys Val Lys Asp Val Ile Thr Asn Cys Lys Ser Cys Lys Glu Ser Gly
785 790 795 800
Asn Lys Cys Lys Thr Glu Cys Lys Thr Lys Cys Lys Asp Glu Cys Glu
805 810 815
Lys Tyr Lys Lys Phe Ile Glu Ala Cys Gly Thr Ala Gly Gly Gly Ile
820 825 830
Gly Thr Ala Gly Ser Pro Trp Ser Lys Arg Trp Asp Gln Ile Tyr Lys
835 840 845
Arg Tyr Ser Lys His Ile Glu Asp Ala Lys Arg Asn Arg Lys Ala Gly
850 855 860
Thr Lys Asn Cys Gly Thr Ser Ser Thr Thr Asn Ala Ala Ala Ser Thr
865 870 875 880
Asp Glu Asn Lys Cys Val Gln Ser Asp Ile Asp Ser Phe Phe Lys His
885 890 895
Leu Ile Asp Ile Gly Leu Thr Thr Pro Ser Ser Tyr Leu Ser Asn Val
900 905 910
Leu Asp Asp Asn Ile Cys Gly Ala Asp Lys Ala Pro Trp Thr Thr Tyr
915 920 925
Thr Thr Tyr Thr Thr Thr Glu Lys Cys Asn Lys Glu Arg Asp Lys Ser
930 935 940
Lys Ser Gln Ser Ser Asp Thr Leu Val Val Val Asn Val Pro Ser Pro
945 950 955 960
Leu Gly Asn Thr Pro Tyr Arg Tyr Lys Tyr Ala Cys Gln Cys Lys Ile
965 970 975
Pro Thr Asn Glu Glu Thr Cys Asp Asp Arg Lys Glu Tyr Met Asn Gln
980 985 990
Trp Ser Cys Gly Ser Ala Arg Thr Met Lys Arg Gly Tyr Lys Asn Asp
995 1000 1005
Asn Tyr Glu Leu Cys Lys Tyr Asn Gly Val Asp Val Lys Pro Thr
1010 1015 1020
Thr Val Arg Ser Asn Ser Ser Lys Leu Asp Ser Gly Arg
1025 1030 1035
<210> 76
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 76
aactacatca agggcgac 18
<210> 77
<211> 21
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 77
cttgttgata ttggtgtcgg t 21
<210> 78
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 78
cacagcgata gcggcaag 18
<210> 79
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 79
gtccagcttg ctggagtt 18
<210> 80
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 80
aactacatca agggcgac 18
<210> 81
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 81
gtccagcttg ctggagtt 18
<210> 82
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 82
aactacatca agggcgac 18
<210> 83
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 83
agcggcgttg gtggtgga 18
<210> 84
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 84
aactacatca agggcgac 18
<210> 85
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 85
gtacttgtac cggtaggg 18
<210> 86
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 86
cacagcgata gcggcaag 18
<210> 87
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 87
gtacttgtac cggtaggg 18
<210> 88
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 88
ctgaccaact gctacaag 18
<210> 89
<211> 19
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 89
ggtccagagg gtacagctt 19
<210> 90
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 90
ctgtccttca tcctgaac 18
<210> 91
<211> 21
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 91
ttcagcgttg ttgtactcgt a 21
<210> 92
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 92
ctgtccttca tcctgaac 18
<210> 93
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 93
gtccagaggg tacagctt 18
<210> 94
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 94
cactctgact ctggcacc 18
<210> 95
<211> 24
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 95
agaggacttc atcttgttgt tggt 24
<210> 96
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 96
ctgtccttca tcctgaac 18
<210> 97
<211> 24
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 97
agaggacttc atcttgttgt tggt 24
<210> 98
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 98
cactctgact ctggcacc 18
<210> 99
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 99
gtccagctta gaggagtt 18
<210> 100
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 100
ctgtccttca tcctgaac 18
<210> 101
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 101
gtccagctta gaggagtt 18
<210> 102
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 102
cactctgact ctggcacc 18
<210> 103
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 103
ggcggcgttg gtggtaga 18
<210> 104
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 104
ctgtccttca tcctgaac 18
<210> 105
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 105
ggcggcgttg gtggtaga 18
<210> 106
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 106
cactctgact ctggcacc 18
<210> 107
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 107
gtacttgtat ccgtgggg 18
<210> 108
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 108
ctgtccttca tcctgaac 18
<210> 109
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 109
gtacttgtat ccgtgggg 18
<210> 110
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 110
cacagcgata gcggcaag 18
<210> 111
<211> 36
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 111
ggtgtcgaag ttgatgtcgg gcagattgcc caggta 36
<210> 112
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 112
cacagcgata gcggcaag 18
<210> 113
<211> 36
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 113
agctgcggcc agattagcgc cctcgtggaa ggacac 36
<210> 114
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 114
cacagcgata gcggcaag 18
<210> 115
<211> 36
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 115
agcgcattca gctgcggcgt tggtcttgat ggagct 36
<210> 116
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 116
cacagcgata gcggcaag 18
<210> 117
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 117
gtccagcttg ctggagtt 18
<210> 118
<211> 36
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 118
gctaatctgg ccgcagctta cccccagaat aagaac 36
<210> 119
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 119
gtccagcttg ctggagtt 18
<210> 120
<211> 36
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 120
gccgcagctg aatgcgctga cgtgaagctg ggcgtg 36
<210> 121
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 121
gtccagcttg ctggagtt 18
<210> 122
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 122
cacagcgata gcggcaag 18
<210> 123
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 123
gtccagcttg ctggagtt 18
<210> 124
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 124
cacagcgata gcggcaag 18
<210> 125
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 125
gtccagcttg ctggagtt 18
<210> 126
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 126
cacagcgata gcggcaag 18
<210> 127
<211> 18
<212> DNA
<213> 人工
<220>
<223> 引物序列
<400> 127
gtccagcttg ctggagtt 18
<210> 128
<211> 640
<212> PRT
<213> 人工序列
<220>
<223> 人工构建体
<400> 128
Asn Tyr Ile Lys Gly Asp Pro Tyr Phe Ala Glu Tyr Ala Thr Lys Leu
1 5 10 15
Ser Phe Ile Leu Asn Pro Ser Asp Ala Asn Asn Pro Ser Gly Glu Thr
20 25 30
Ala Asn His Asn Asp Glu Ala Cys Asn Cys Asn Glu Ser Gly Ile Ser
35 40 45
Ser Val Gly Gln Ala Gln Thr Ser Gly Pro Ser Ser Gln Lys Thr Cys
50 55 60
Ile Thr His Ser Ser Ile Lys Thr Asn Lys Lys Lys Glu Cys Lys Asp
65 70 75 80
Val Lys Leu Gly Val Arg Glu Asn Asp Lys Asp Leu Lys Ile Cys Val
85 90 95
Ile Glu Asp Thr Ser Leu Ser Gly Val Asp Asn Cys Cys Cys Gln Asp
100 105 110
Leu Leu Gly Ile Leu Gln Glu Asn Cys Ser Asp Asn Lys Arg Gly Ser
115 120 125
Ser Ser Asn Asp Ser Cys Asp Asn Lys Asn Gln Asp Glu Cys Gln Lys
130 135 140
Lys Leu Glu Lys Val Phe Ala Ser Leu Thr Asn Gly Tyr Lys Cys Asp
145 150 155 160
Lys Cys Lys Ser Gly Thr Ser Arg Ser Lys Lys Lys Trp Ile Trp Lys
165 170 175
Lys Ser Ser Gly Asn Glu Glu Gly Leu Gln Glu Glu Tyr Ala Asn Thr
180 185 190
Ile Gly Leu Pro Pro Arg Thr Gln Ser Leu Tyr Leu Gly Asn Leu Pro
195 200 205
Lys Leu Glu Asn Val Cys Glu Asp Val Lys Asp Ile Asn Phe Asp Thr
210 215 220
Lys Glu Lys Phe Leu Ala Gly Cys Leu Ile Val Ser Phe His Glu Gly
225 230 235 240
Lys Asn Leu Lys Lys Arg Tyr Pro Gln Asn Lys Asn Ser Gly Asn Lys
245 250 255
Glu Asn Leu Cys Lys Ala Leu Glu Tyr Ser Phe Ala Asp Tyr Gly Asp
260 265 270
Leu Ile Lys Gly Thr Ser Ile Trp Asp Asn Glu Tyr Thr Lys Asp Leu
275 280 285
Glu Leu Asn Leu Gln Asn Asn Phe Gly Lys Leu Phe Gly Lys Tyr Ile
290 295 300
Lys Lys Gln Asn Thr Ala Glu Gln Asp Thr Ser Tyr Ser Ser Leu Asp
305 310 315 320
Glu Leu Arg Glu Ser Trp Trp Asn Thr Asn Lys Lys Tyr Ile Trp Thr
325 330 335
Ala Met Lys His Gly Ala Glu Met Gln Ile Thr Thr Cys Asn Ala Asp
340 345 350
Gly Ser Val Thr Gly Ser Gly Ser Ser Cys Asp Asp Ile Pro Thr Ile
355 360 365
Asp Leu Ile Pro Gln Tyr Leu Arg Phe Leu Gln Glu Trp Val Glu Asn
370 375 380
Phe Cys Glu Gln Arg Gln Ala Lys Val Lys Asp Val Ile Thr Asn Cys
385 390 395 400
Lys Ser Cys Lys Glu Ser Gly Asn Lys Cys Lys Thr Glu Cys Lys Thr
405 410 415
Lys Cys Lys Asp Glu Cys Glu Lys Tyr Lys Lys Phe Ile Glu Ala Cys
420 425 430
Gly Thr Ala Gly Gly Gly Ile Gly Thr Ala Gly Ser Pro Trp Ser Lys
435 440 445
Arg Trp Asp Gln Ile Tyr Lys Arg Tyr Ser Lys His Ile Glu Asp Ala
450 455 460
Lys Arg Asn Arg Lys Ala Gly Thr Lys Asn Cys Gly Thr Ser Ser Thr
465 470 475 480
Thr Asn Ala Ala Ala Ser Thr Asp Glu Asn Lys Cys Val Gln Ser Asp
485 490 495
Ile Asp Ser Phe Phe Lys His Leu Ile Asp Ile Gly Leu Thr Thr Pro
500 505 510
Ser Ser Tyr Leu Ser Asn Val Leu Asp Asp Asn Ile Cys Gly Ala Asp
515 520 525
Lys Ala Pro Trp Thr Thr Tyr Thr Thr Tyr Thr Thr Thr Glu Lys Cys
530 535 540
Asn Lys Glu Arg Asp Lys Ser Lys Ser Gln Ser Ser Asp Thr Leu Val
545 550 555 560
Val Val Asn Val Pro Ser Pro Leu Gly Asn Thr Pro Tyr Arg Tyr Lys
565 570 575
Tyr Ala Cys Gln Cys Lys Ile Pro Thr Asn Glu Glu Thr Cys Asp Asp
580 585 590
Arg Lys Glu Tyr Met Asn Gln Trp Ser Cys Gly Ser Ala Arg Thr Met
595 600 605
Lys Arg Gly Tyr Lys Asn Asp Asn Tyr Glu Leu Cys Lys Tyr Asn Gly
610 615 620
Val Asp Val Lys Pro Thr Thr Val Arg Ser Asn Ser Ser Lys Leu Asp
625 630 635 640
<210> 129
<211> 116
<212> PRT
<213> 人工序列
<220>
<223> 人工构建体
<400> 129
Gly Ala Met Val Asp Thr Leu Ser Gly Leu Ser Ser Glu Gln Gly Gln
1 5 10 15
Ser Gly Asp Met Thr Ile Glu Glu Asp Ser Ala Thr His Ile Lys Phe
20 25 30
Ser Lys Arg Asp Glu Asp Gly Lys Glu Leu Ala Gly Ala Thr Met Glu
35 40 45
Leu Arg Asp Ser Ser Gly Lys Thr Ile Ser Thr Trp Ile Ser Asp Gly
50 55 60
Gln Val Lys Asp Phe Tyr Leu Tyr Pro Gly Lys Tyr Thr Phe Val Glu
65 70 75 80
Thr Ala Ala Pro Asp Gly Tyr Glu Val Ala Thr Ala Ile Thr Phe Thr
85 90 95
Val Asn Glu Gln Gly Gln Val Thr Val Asn Gly Lys Ala Thr Lys Gly
100 105 110
Asp Ala His Ile
115
<210> 130
<211> 13
<212> PRT
<213> 人工序列
<220>
<223> 人工Spytag序列
<400> 130
Ala His Ile Val Met Val Asp Ala Tyr Lys Pro Thr Lys
1 5 10
<210> 131
<211> 8
<212> PRT
<213> 人工序列
<220>
<223> 人工Spytag序列
<400> 131
Ala His Ile Val Met Val Asp Ala
1 5
<210> 132
<211> 10
<212> PRT
<213> 人工序列
<220>
<223> The beta-strand of CnaB2
<400> 132
Ala Thr His Ile Lys Phe Ser Lys Arg Asp
1 5 10
<210> 133
<211> 104
<212> PRT
<213> 人工序列
<220>
<223> SpyLigase序列
<400> 133
His His His His His His Asp Tyr Asp Gly Gln Ser Gly Asp Gly Lys
1 5 10 15
Glu Leu Ala Gly Ala Thr Met Glu Leu Arg Asp Ser Ser Gly Lys Thr
20 25 30
Ile Ser Thr Trp Ile Ser Asp Gly Gln Val Lys Asp Phe Tyr Leu Tyr
35 40 45
Pro Gly Lys Tyr Thr Phe Val Glu Thr Ala Ala Pro Asp Gly Tyr Glu
50 55 60
Val Ala Thr Ala Ile Thr Phe Thr Val Asn Glu Gln Gly Gln Val Thr
65 70 75 80
Val Asn Gly Lys Ala Thr Lys Gly Gly Ser Gly Gly Ser Gly Gly Ser
85 90 95
Gly Glu Asp Ser Ala Thr His Ile
100
<210> 134
<211> 16
<212> PRT
<213> 人工序列
<220>
<223> 异肽Spy0128序列
<400> 134
Thr Asp Lys Asp Met Thr Ile Thr Phe Thr Asn Lys Lys Asp Ala Glu
1 5 10 15
<210> 135
<211> 282
<212> PRT
<213> 人工序列
<220>
<223> Split-Spy0128序列
<400> 135
Ala Thr Thr Val His Gly Glu Thr Val Val Asn Gly Ala Lys Leu Thr
1 5 10 15
Val Thr Lys Asn Leu Asp Leu Val Asn Ser Asn Ala Leu Ile Pro Asn
20 25 30
Thr Asp Phe Thr Phe Lys Ile Glu Pro Asp Thr Thr Val Asn Glu Asp
35 40 45
Gly Asn Lys Phe Lys Gly Val Ala Leu Asn Thr Pro Met Thr Lys Val
50 55 60
Thr Tyr Thr Asn Ser Asp Lys Gly Gly Ser Asn Thr Lys Thr Ala Glu
65 70 75 80
Phe Asp Phe Ser Glu Val Thr Phe Glu Lys Pro Gly Val Tyr Tyr Tyr
85 90 95
Lys Val Thr Glu Glu Lys Ile Asp Lys Val Pro Gly Val Ser Tyr Asp
100 105 110
Thr Thr Ser Tyr Thr Val Gln Val His Val Leu Trp Asn Glu Glu Gln
115 120 125
Gln Lys Pro Val Ala Thr Tyr Ile Val Gly Tyr Lys Glu Gly Ser Lys
130 135 140
Val Pro Ile Gln Phe Lys Asn Ser Leu Asp Ser Thr Thr Leu Thr Val
145 150 155 160
Lys Lys Lys Val Ser Gly Thr Gly Gly Asp Arg Ser Lys Asp Phe Asn
165 170 175
Phe Gly Leu Thr Leu Lys Ala Asn Gln Tyr Tyr Lys Ala Ser Glu Lys
180 185 190
Val Met Ile Glu Lys Thr Thr Lys Gly Gly Gln Ala Pro Val Gln Thr
195 200 205
Glu Ala Ser Ile Asp Gln Leu Tyr His Phe Thr Leu Lys Asp Gly Glu
210 215 220
Ser Ile Lys Val Thr Asn Leu Pro Val Gly Val Asp Tyr Val Val Thr
225 230 235 240
Glu Asp Asp Tyr Lys Ser Glu Lys Tyr Thr Thr Asn Val Glu Val Ser
245 250 255
Pro Gln Asp Gly Ala Val Lys Asn Ile Ala Gly Asn Ser Thr Glu Gln
260 265 270
Glu Thr Ser Thr Asp Lys Asp Met Thr Ile
275 280
<210> 136
<211> 93
<212> PRT
<213> 人工序列
<220>
<223> SpyCatcher-delta-N序列
<400> 136
Glu Asp Ser Ala Thr His Ile Lys Phe Ser Lys Arg Asp Glu Asp Gly
1 5 10 15
Lys Glu Leu Ala Gly Ala Thr Met Glu Leu Arg Asp Ser Ser Gly Lys
20 25 30
Thr Ile Ser Thr Trp Ile Ser Asp Gly Gln Val Lys Asp Phe Tyr Leu
35 40 45
Tyr Pro Gly Lys Tyr Thr Phe Val Glu Thr Ala Ala Pro Asp Gly Tyr
50 55 60
Glu Val Ala Thr Ala Ile Thr Phe Thr Val Asn Glu Gln Gly Gln Val
65 70 75 80
Thr Val Asn Gly Lys Ala Thr Lys Gly Asp Ala His Ile
85 90
<210> 137
<211> 13
<212> PRT
<213> 人工序列
<220>
<223> 反向spytaq序列
<400> 137
Lys Thr Pro Lys Tyr Ala Asp Val Met Val Ile His Ala
1 5 10

Claims (34)

1.一种多肽，其包含i)第一多肽结构域，其是嵌合抗原受体(CAR)的跨膜结构域和胞内结构域，以及ii)选自VAR2CSA多肽或分割蛋白结合系统的肽部分的第二胞外多肽结构域。
2.权利要求1的多肽，其中包含所述CAR和分割蛋白结合系统的肽部分的多肽可以在体外或体内与分割蛋白标记的肿瘤靶向剂例如抗体组合。
3.一种多肽，其包含i)第一多肽结构域，其是VAR2CSA多肽或其它肿瘤靶向剂，例如单链scFv，和ii)选自分割蛋白结合系统的肽部分、单结构域抗体例如scFv或纳米抗体中的一个或多个的第二多肽结构域。
4.权利要求1或2中任一项的多肽，其中所述分割蛋白结合系统的一个肽部分选自K-Tag(SEQ ID NO：132)，SpyCatcher(SEQ ID NO：129)，SpyCatcher-ΔN(SEQ ID NO：136)，SpyTag(SEQ ID NO：130)，最小Spytag序列(SEQ ID NO：131)，split-Spy0128(SEQ ID NO：135)，异肽Spy0128(SEQ ID NO：134)或其任何反向序列或其具有至少约80％，例如至少约82,84,86,88,90,92,94,96,98或99％的序列同一性的变体。
5.权利要求1-3中任一项的多肽，其中所述多肽还包含一个或多个接头或铰链序列，例如GC接头。
6.权利要求1-4中任一项的多肽，其中所述VAR2CSA多肽由SEQ ID NO：55或SEQ ID NO：56或其具有结合硫酸软骨素A(CSA)的能力的片段或变体组成，所述硫酸软骨素A(CSA)可以被呈现在蛋白聚糖(CSPG)上。
7.权利要求1-5中任一项的多肽，其中所述VAR2CSA多肽是VAR2CSA的片段，其由以下的连续氨基酸序列组成：
a)ID1，和
b)DBL2Xb，以及任选的
c)ID2a。
8.权利要求1-6中任一项的多肽，其中所述VAR2CSA多肽以通过小于100nM，例如小于80nM，例如小于70nM，例如小于60nM，例如小于50nM，例如小于40nM，例如小于30nM，例如小于26nM，例如小于24nM，例如小于22nM，例如小于20nM，例如小于18nM，例如小于16nM，例如小于14nM，例如小于12nM，例如小于10nM，例如小于9nM，例如小于8nM，例如小于7nM，例如小于6nM，或小于4nM的KD测量的亲和力结合蛋白聚糖(CSPG)上的硫酸软骨素A(CSA)。
9.权利要求1-7中任一项的多肽，其中所述VAR2CSA多肽包含与SEQ ID NO：1的1-577，SEQ ID NO：3的1-592，SEQ ID NO：4的1-579，SEQ ID NO：5的1-576，SEQ ID NO：10的1-586，SEQ ID NO：11的1-579，SEQ ID NO：29的1-565，SEQ ID NO：34的1-584，SEQ ID NO：36的1-569，SEQ ID NO：37的1-575，SEQ ID NO：38的1-592，SEQ ID NO：41的1-603，SEQ ID NO：43的1-588，SEQ ID NO：44的1-565，SEQ ID NO：45的1-589，SEQ ID NO：48的1-573，SEQ IDNO：53的1-583，SEQ ID NO：54的1-569或SEQ ID NO：128的1-640的任何一个氨基酸序列具有至少70％序列同一性的氨基酸序列。
10.权利要求1-8中任一项的多肽，其中所述VAR2CSA多肽包含与SEQ ID NO：1的578-640，SEQ ID NO：3的593-656，SEQ ID NO：4的580-643，SEQ ID NO：5的577-640，SEQ ID NO：10的587-650，SEQ ID NO：11的580-643，SEQ ID NO：29的566-628，SEQ ID NO：34的585-647，SEQ ID NO：36的570-632，SEQ ID NO：37的576-639，SEQ ID NO：38的593-655，SEQ IDNO：41的604-667，SEQ ID NO：43的589-652，SEQ ID NO：44的566-628，SEQ ID NO：45的590-653，SEQ ID NO：48的574-637，SEQ ID NO：53的584-646或SEQ ID NO：54的570-632的氨基酸序列具有至少70％序列同一性的氨基酸序列。
11.权利要求1-9中任一项的多肽，其中所述VAR2CSA多肽包含与SEQ ID NO：2,6,8,9,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,30,31,32,33,35,39,40,42,46,47,49,50,51,52,或128的氨基酸序列具有至少70％序列同一性的氨基酸序列。
12.权利要求1-10中任一项的多肽，其中所述VAR2CSA多肽由与SEQ ID NO：1的1-577，SEQ ID NO：3的1-592，SEQ ID NO：4的1-579，SEQ ID NO：5的1-576，SEQ ID NO：10的1-586，SEQ ID NO：11的1-579，SEQ ID NO：29的1-565，SEQ ID NO：34的1-584，SEQ ID NO：36的1-569，SEQ ID NO：37的1-575，SEQ ID NO：38的1-592，SEQ ID NO：41的1-603，SEQ ID NO：43的1-588，SEQ ID NO：44的1-565，SEQ ID NO：45的1-589，SEQ ID NO：48的1-573，SEQ IDNO：53的1-583，SEQ ID NO：54的1-569或SEQ ID NO：128的1-640的任何一个氨基酸序列具有至少70％序列同一性的氨基酸序列组成。
13.权利要求1-11中任一项的多肽，其中所述VAR2CSA多肽由选自SEQ ID NO：1,3-5,10,11,29,34,36-38,41,43-45,48,53,54和128的氨基酸序列组成。
14.权利要求1-12中任一项的多肽，其中所述VAR2CSA多肽由长度小于700个氨基酸，例如小于690个氨基酸，例如小于680个氨基酸，例如小于670个氨基酸，例如小于660个氨基酸，例如小于650个氨基酸，例如小于640个氨基酸，例如小于630个氨基酸，例如小于620个氨基酸，例如小于610个氨基酸，例如小于600个氨基酸，例如小于590个氨基酸，例如小于580个氨基酸，例如小于570个氨基酸的氨基酸序列组成。
15.权利要求1-13中任一项的多肽，其中所述VAR2CSA多肽具有在非还原条件下在SDS-PAGE上的小于约100kDa的分子量。
16.权利要求1-14中任一项的多肽，其中所述VAR2CSA多肽是非糖基化的。
17.权利要求1-15中任一项的多肽，其中嵌合抗原受体(CAR)的所述跨膜结构域和/或胞内结构域来源于第1代、第2代或第3代CAR。
18.权利要求1-15中任一项的多肽，其中包含嵌合抗原受体(CAR)的跨膜结构域和胞内结构域作为第一多肽结构域的所述多肽还包含其它调节元件或自杀基因系统，例如诱导型半胱天冬酶-9(iCASP9)或睡美人(SB)转座子系统。
19.权利要求1-16中任一项的多肽，其中嵌合抗原受体(CAR)的所述跨膜结构域和/或胞内结构域包含CD28和/或CD3ζ信号转导结构域。
20.权利要求1-3中任一项的多肽，其中嵌合抗原受体(CAR)的所述跨膜结构域和/或胞内结构域是如WO2014127261，WO13126729，WO13126733，WO14055442，WO14055771，WO2014039523，WO14179759和WO14138704中任一个所描述的。
21.一种分离的核酸分子，其包含编码权利要求1-19的多肽的核苷酸序列。
22.一种表达载体，其包含权利要求21的分离的核酸分子。
23.一种细胞，其包含权利要求21的载体。
24.权利要求21的细胞，其中所述细胞选自αβT细胞，γδT细胞，自然杀伤(NK)细胞，细胞毒性T淋巴细胞(CTL)，调节性T细胞或其任何组合。
25.一种双组分治疗剂，其包含
a)第一组分，其是多肽，所述多肽包含i)第一多肽结构域，其是嵌合抗原受体(CAR)的跨膜结构域和胞内结构域，以及ii)第二多肽结构域，其是连接至所述CAR的胞外部分的分割蛋白结合系统的肽部分；和
b)第二组分，其是多肽，所述多肽包含i)第一多肽结构域，其是VAR2CSA多肽或任何其它肿瘤靶向剂，例如抗体或scFv，以及ii)第二多肽结构域，其是分割蛋白结合系统的肽部分；任选地还包含iii)单结构域抗体，例如scFv或纳米抗体，
其中由组分a)和b)定义的分割蛋白结合系统的所述肽部分自发形成不可逆的异肽键。
26.双组分治疗剂，其中所述多肽如权利要求1-19中任一项所定义。
27.一种治疗或预防受试者中的与例如癌症中CSA的表达例如不适当表达相关的任何适应症或病况的发生的方法，所述方法包括：(a)在促进活化的条件下培养细胞毒性淋巴细胞群；(b)用编码多肽的载体转染(a)的淋巴细胞群，所述多肽包含i)第一多肽结构域，其是嵌合抗原受体(CAR)的跨膜结构域和胞内结构域，以及ii)第二多肽结构域，其选自VAR2CSA多肽或任何其它靶向剂，例如抗体；(c)将治疗或预防有效量的(b)的经转染的淋巴细胞施用于具有所述适应症或处于具有所述适应症的风险中的受试者；其中包含CAR的结构域的多肽具有针对靶向部分的结合特异性，或可被靶向部分结合；从而治疗或预防所述受试者的所述适应症。
28.一种治疗或预防受试者中的与例如癌症中CSA的表达例如不适当表达相关的任何适应症或病况的发生的方法，所述方法包括：(a)在促进活化的条件下培养细胞毒性淋巴细胞群；(b)用编码多肽的载体转染(a)的淋巴细胞群，所述多肽包含i)第一多肽结构域，其是嵌合抗原受体(CAR)的跨膜结构域和胞内结构域，以及ii)第二多肽结构域，其是连接至所述CAR的胞外部分的分割蛋白结合系统的肽部分；(c)将治疗或预防有效量的(b)的经转染的淋巴细胞施用于具有所述适应症或处于具有所述适应症的风险中的受试者；和(d)施用多肽，所述多肽包含i)第一多肽结构域，其为VAR2CSA多肽或其它靶向剂，例如抗体，和ii)第二多肽结构域，其是分割蛋白结合系统的肽部分，其中在所使用的两种不同的多肽中定义的分割蛋白结合系统的所述肽部分自发形成不可逆的异肽键；从而治疗或预防所述受试者的所述适应症。
29.权利要求1-19中任一项的多肽或权利要求24-25中任一项的双组分治疗剂，其用作药物。
30.一种药物组合物，其包含权利要求1-19中任一项的多肽或权利要求24-25中任一项的双组分治疗剂。
31.权利要求1-19中任一项的多肽或权利要求24-25中任一项的双组分治疗剂用于制备药物的用途。
32.权利要求1-19中任一项的多肽或权利要求24-25中任一项的双组分治疗剂或权利要求29的药物组合物，用于治疗与例如癌症中涉及CSA的表达例如不适当表达的病况相关的任何适应症。
33.权利要求31的多肽或双组分治疗剂或权利要求26或27的方法，其中所述癌症选自所有表达CSA的恶性肿瘤，包括癌(包括但不限于乳腺癌，胰腺癌，卵巢癌，子宫内膜癌，肝细胞癌，肺癌，结肠癌，前列腺癌，宫颈癌，睾丸癌，基底细胞皮肤癌，透明细胞肾细胞癌，头颈鳞状细胞癌，皮肤鳞状细胞癌，外阴角质化鳞状细胞癌和外阴基底细胞癌)，肉瘤(包括但不限于纤维肉瘤，去分化的软骨和脂肪肉瘤，平滑肌肉瘤，脂肪肉瘤，粘液样脂肪肉瘤，子宫体平滑肌肉瘤，骨肉瘤，尤文肉瘤和横纹肌肉瘤，滑膜肉瘤，孤立性纤维瘤)，造血系统癌(包括但不限于慢性淋巴细胞白血病(CLL)，急性淋巴细胞白血病(ALL)，急性骨髓性白血病(AML)，B细胞、T细胞和大颗粒淋巴瘤)，神经上皮组织的肿瘤，例如但不限于星形细胞瘤(多形性黄色星形细胞瘤，纤维性星形细胞瘤，间变性星形细胞瘤，多形性成胶质细胞瘤)，少突神经胶质瘤，室管膜瘤，脉络丛肿瘤，少突星形细胞瘤，神经胶质肉瘤，神经节胶质细胞瘤，视网膜母细胞瘤，神经细胞瘤，神经母细胞瘤(成感觉神经细胞瘤和神经节成神经细胞瘤)，髓母细胞瘤，非典型畸胎样横纹肌样瘤和所有类型的神经内分泌癌以及任何其它表达CSA的癌症。
34.权利要求1-19中任一项的多肽或权利要求24-25中任一项的双组分治疗剂或权利要求29的药物组合物，其与任何其它合适的抗癌剂组合。
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CN114206944A (zh) * 2019-07-12 2022-03-18 哥本哈根大学 用于治疗cmv感染相关疾病的融合毒素蛋白
WO2022016375A1 (zh) * 2020-07-21 2022-01-27 深圳先进技术研究院 异位子宫内膜识别多肽及其衍生物和应用
CN114075298A (zh) * 2022-01-07 2022-02-22 广州中科蓝华生物科技有限公司 一种索烃化的var2csa重组蛋白及其制备方法和应用
CN114957432A (zh) * 2022-06-06 2022-08-30 山东大学 一种能特异性识别肝素类结构的蛋白探针var2hp及其编码基因与应用
CN114957432B (zh) * 2022-06-06 2023-07-21 山东大学 一种能特异性识别肝素类结构的蛋白探针var2hp及其编码基因与应用
WO2024120537A1 (zh) * 2022-12-09 2024-06-13 中山大学肿瘤防治中心（中山大学附属肿瘤医院、中山大学肿瘤研究所） 一种重组var2csa及其在肿瘤型硫酸软骨素蛋白聚糖检测中的应用
WO2024120538A1 (zh) * 2022-12-09 2024-06-13 中山大学肿瘤防治中心（中山大学附属肿瘤医院、中山大学肿瘤研究所） 一种var2csa重组蛋白及其制备方法和应用

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