CN110582295A - 多种疟疾红前期抗原及其在宿主中引发保护性免疫应答的用途 - Google Patents

多种疟疾红前期抗原及其在宿主中引发保护性免疫应答的用途 Download PDF

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CN110582295A
CN110582295A CN201880021704.3A CN201880021704A CN110582295A CN 110582295 A CN110582295 A CN 110582295A CN 201880021704 A CN201880021704 A CN 201880021704A CN 110582295 A CN110582295 A CN 110582295A
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R·阿米诺
P·沙尔诺
A-S·贝尼翁
C·布兰克
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Abstract

本发明涉及疟原虫抗原多肽，该疟原虫抗原多肽通过使用特别设计的功能性免疫筛选方法进行鉴定。特别地，本发明涉及疟原虫的抗原多肽，其中，所述抗原多肽显示出保护作用，特别是在寄主中引发针对疟原虫子孢子攻击的保护性免疫应答。本发明涉及化合物的组合物，该化合物的组合物包含至少两种不同的活性成分，其中，每种活性成分由疟原虫寄生虫的抗原多肽、编码抗原多肽的多核苷酸或表达疟原虫寄生虫的此种抗原多肽的载体(特别是病毒载体，特别是慢病毒)组成，其中，一种抗原多肽是环子孢子蛋白(CSP)或其多肽衍生物，另一种抗原多肽是蛋白Ag40(11‑09)或蛋白Ag45(11‑10)。

Description

多种疟疾红前期抗原及其在宿主中引发保护性免疫应答的 用途
技术领域
本发明涉及使用特别设计的功能性免疫筛选实验(functional immunizationscreening assay)鉴别的疟原虫(Plasmodium)抗原多肽。特别地，本发明涉及疟原虫(malaria parasite)的抗原多肽，其中，所述抗原多肽在寄主中针对疟原虫子孢子(sporozoite)的攻击显示出保护作用，特别是引发保护性免疫应答。因此，此种被鉴别的抗原多肽可以构成适用于候选疫苗设计的活性成分，特别是适用于人类寄主的疫苗。
背景技术
在过去的15年中，疟疾控制措施使此种蚊媒病(mosquito-borne disease)造成的全球死亡人数减少了48％。尽管死亡率显著下降，但WHO估计约有2.15亿的疟疾临床发作(clinical episode)，导致2015年死亡人数超过40万。实际的疟疾控制计划主要依靠使用杀虫剂和抗疟原虫药物，但耐药蚊子和寄生虫的出现和传播使这些干预措施的效力受到威胁1。在此种情况下，有效的疟疾疫苗可以成为控制并最终消除疟疾的重要辅助工具。
据悉，自60年代以来，使用经辐照的子孢子的多种免疫可以引发针对疟疾感染的无菌保护。然而，在过去的50年中，仅鉴别了少数保护性抗原，但它们中没有任何一种(无论是单独还是组合)能与经辐照的寄生虫引起的强力保护(robust protection)相配。
最先进的疟疾疫苗RTS,S(Mosquirix，GSK)靶向恶性疟原虫(Plasmodiumfalciparum)环子孢子蛋白(CSP，circumsporozoite protein)，CSP是子孢子的主要表面蛋白，子孢子是感染性蚊虫叮咬期间接种到皮肤的游动期(motile stage)。此种亚单位疫苗(subunit vaccine)使非洲婴儿和儿童的疟疾临床病例减少了26％至36％2。此种局部保护主要与高滴度的抗CSP抗体相关3，尽管显著，但远未达到WHO疟疾疫苗路线图(WHO malariavaccine road map)所制定的标准，该路线图宣告开发抗临床疟疾功效为至少75％的疫苗，理想地针对发病率、死亡率和寄生虫传播4。
另一方面，活的经辐照的子孢子可以侵入但在肝期(liver-stage)早期在肝细胞内被抑制，从而产生针对同源子孢子攻击的无菌免疫5。遗憾的是，在贫穷的热带国家，这些活寄生虫的生产、储存和递送相关的技术和经济障碍仍阻碍它们用于大规模疫苗接种。此种无菌保护似乎主要依赖于CD8+T细胞，因为几个实验模型中CD8+T细胞的耗竭消除了无菌免疫，然而，引起此种强力保护的抗原的身份尚不清楚6。迄今为止，在由红前(PE，pre-erythrocytic)期、子孢子和随后的肝期表达的数千种可能的蛋白质中，已知保护性抗原的数量极为有限；而且这些抗原在人类中仅提供了比单独的CSP制剂或多抗原(multi-antigenic)制剂更弱的保护作用7。迄今为止，尽管筛选了数千种PE肽、小基因(mini-gene)和基因8-10，但从减毒(attenuated)活子孢子中鉴别新的保护性抗原的尝试尚未得到合适的候选物。
发明内容
作为本发明的活性成分提供的抗原多肽
为了鉴别关键的保护性抗原，本发明提供了基于慢病毒的免疫筛选，其设计用于选择能够保护易感(susceptible)小鼠免受严重的子孢子攻击的疟原虫保守抗原(plasmodial-conserved antigen)。如实施例所示，使用的此功能性筛选，发明人从55种测试抗原中鉴别出8种保护性抗原，包括已知的候选疫苗CSP和TRAP。值得注意的是，发明人表明：7种灭菌抗原的组合保护了超过85％(18/21)的受攻击动物，而在CSP免疫组中保护了5％(1/20)。此外，核心的5种抗原被确定为该有效无菌保护的来源。这些研究结果适用于源自感染人类的疟原虫寄生虫的抗原，为开发多抗原的第二代PE疟疾疫苗铺平了道路。
因此，本发明涉及化合物的组合物，该组合物包含至少两种不同的活性成分，其中，每种活性成分由疟原虫寄生虫的抗原多肽、编码抗原多肽的多核苷酸或载体(特别是病毒载体，特别是慢病毒载体)组成；其中，该载体表达疟原虫寄生虫的此种抗原多肽；其中，一种抗原多肽是环子孢子蛋白(CSP)或它的多肽衍生物，另一种抗原多肽是蛋白Ag40(11-09)(其具有SEQ ID NO：67、68、70、71、73或74的序列之一)或它的多肽衍生物，或蛋白Ag45(11-10)(其具有SEQ ID NO：76、77、79、80、82或83的序列之一或彼此独立)或它的多肽衍生物，前提是组合物中每种多肽衍生物保持衍生其的抗原的保护性。
在本发明的一个具体实施方式中，该组合物进一步包含以下物质作为活性成分：一种以上疟原虫寄生虫的抗原多肽、编码抗原多肽的多核苷酸，或载体(特别是病毒载体，特别是慢病毒载体)；其中，该载体表达疟原虫寄生虫的此种抗原多肽；其中，每种抗原多肽选自：血小板反应蛋白相关匿名蛋白(TRAP，thrombospondin related anonymousprotein)、半胱氨酸蛋白酶抑制剂(ICP)、金属肽酶(Bergheilysin/Falcilysin)、GPI锚定蛋白(GPI-anchored protein)P113、成孔样蛋白(pore-forming like protein)SPECT2，或者分别且彼此独立地，任何这些抗原多肽的多肽衍生物；其中，本发明的化合物的组合物中的所述多肽衍生物保持衍生其的抗原的保护性。
因此，该化合物的组合物包含2、3、4、5、6、7或8种抗原，或编码此种抗原多肽的多核苷酸，或载体(特别是病毒载体，特别是慢病毒载体)，其中，该载体表达此种抗原；或可选地，该化合物的组合物由2、3、4、5、6、7或8种抗原的组合或表达此种抗原的病毒载体(特别是慢病毒载体)组成。在一个具体实施方式中，其中，如本文所公开的，该化合物的组合物包含至少3种抗原或表达至少3种抗原的病毒载体，这些抗原至少由环子孢子蛋白(CSP)、蛋白Ag40(11-09)、蛋白Ag45(11-10)和/或它们的衍生物组成。
在本发明的一个具体实施方式中，该化合物的组合物是一组不同的活性成分，其中，每种活性成分由疟原虫寄生虫的抗原多肽或编码此种抗原多肽的多核苷酸组成，或者活性成分由表达疟原虫寄生虫的此种抗原多肽的载体(特别是病毒载体，特别是慢病毒载体)组成；其中，所述活性成分的组包括：疟原虫寄生虫的PE阶段抗原，或表达疟原虫寄生虫的PE阶段抗原的病毒载体(特别是慢病毒载体)，该PE阶段抗原包括：环子孢子蛋白(CSP)、血小板反应蛋白相关匿名蛋白(TRAP)、半胱氨酸蛋白酶抑制剂(ICP)、金属肽酶(Bergheilysin/Falcilysin)、GPI锚定蛋白P113、成孔样蛋白SPECT2、蛋白Ag40(11-09)(其具有SEQ ID NO：67、68、70、71、73或74的序列)或它的变体、蛋白Ag45(11-10)(其具有SEQID NO：76、77、79、80、82或83的序列)或它的变体，或任何这些抗原多肽的多肽衍生物，其中，在该化合物的组合物中，所述多肽衍生物保持衍生其的抗原的保护性。
根据本发明，无论所述成分是作为(天然、重组或合成的)多肽、修饰或未修饰的多核苷酸(如RNA和DNA分子)或作为能够表达所述抗原多肽的载体(特别是病毒载体，更特别是慢病毒载体)提供用于施用，所述成分被描述为不同的“活性成分”；这意味着，它们各自引发针对寄生虫的免疫应答，或者它们定性和/或定量地调节并且特别是扩大和/或加强免疫应答；在寄主中，该免疫应答由该化合物的组合物提供或表达的其它抗原多肽引起，因此对该组合引起的定性和/或定量免疫应答具有其自身的活性或作用；以此种方式，该化合物的组合物引发针对疟原虫感染或针对寄生虫诱导的病症或疾病的保护性应答。除了作为不同的活性成分之外，本文定义的抗原多肽统称为引发针对疟原虫感染或针对寄生虫诱导的病症或疾病的保护性免疫应答的活性成分。
表述“载体”涉及适用于将化合物的组合物的编码抗原多肽的多核苷酸递送至施用这些载体的寄主细胞的生物或化学实体。载体是本领域公知的，可以是病毒载体(如腺病毒载体，特别是使用黑猩猩腺病毒制备的载体)，使用改良痘苗病毒、麻疹病毒或黄热病病毒获得的载体。在本领域中，从这些病毒获得的载体以使本领域技术人员能够制备它们的方式公开。可选地和优选地，慢病毒载体适用于制备本发明的化合物的组合物，特别是使用感染人的慢病毒获得的载体，或者取决于寻求的保护寄主的感染动物的慢病毒的载体。本文公开了此类慢病毒的实施例，本发明特别涉及HIV载体(特别是HIV-1载体)的用途。本文提供了构建HIV-1载体的细节，提供关于这方面的每个公开的实施方式旨在与关于抗原多肽公开的每个实施方式一起应用。
表述“疟原虫寄生虫”和“疟疾寄生虫”在本申请中可互换使用。它们指每个和所有形式的寄生虫，该寄生虫与哺乳动物(特别是人类寄主)中寄生虫周期的各个阶段相关，尤其包括：子孢子，特别是接种到寄主皮肤中且接种后存在于血流中的子孢子，或者在肝细胞(肝期)中发育的子孢子；裂殖子，尤其包括肝细胞中产生的裂殖子和红细胞中产生的裂殖子，或在红细胞(红内期，blood-stage)中发育的裂殖子。这些不同形式的寄生虫通过多种特异性抗原来表征，许多抗原是本领域公知和确定的，一些抗原仍然未知且尚未赋予生物学功能。根据感染阶段，抗原通常可以参照它们的表达进行分组或分类。根据本公开的疟原虫寄生虫包括感染人类寄主的寄生虫和感染非人类哺乳动物(特别是啮齿动物，特别是小鼠)的寄生虫。因此，恶性疟原虫、间日疟原虫(Plasmodium vivax)、约氏疟原虫(Plasmodium yoelii)和伯氏疟原虫(Plasmodium berghei)是这些寄生虫的特定实例。食蟹猴疟原虫(Plasmodium cynomolgi)和诺氏疟原虫(Plasmodium knowlesi)主要对猕猴具有感染性，但也可引起人类感染。
根据本发明，表述“抗原多肽”指多肽，该多肽可以是疟原虫寄生虫的天然抗原，或疟原虫寄生虫(特别是伯氏疟原虫(P.berghei)、食蟹猴疟原虫(P.cynomolgi))或感染人类的疟原虫寄生虫(例如恶性疟原虫(P.falciparum)或间日疟原虫(P.vivax))的密码子优化(codon-optimized)或非密码子优化的基因的表达产物。本申请还涉及称为“它(们)的多肽衍生物”的此种抗原多肽的修饰形式，它可以是寄生虫的天然抗原的片段，特别是此种天然抗原的截短形式或经过翻译后修饰的它的修饰形式。衍生多肽具有足以提供一个以上表位的氨基酸序列，并且它保持保护性，使得当它被包含在本发明的化合物的组合物中时，它具有衍生其的和/或表现出此种保护性的抗原多肽的保护活性。甚至可以用衍生物改善参考抗原的保护性。本文公开的抗原衍生物的各种实例显示于实施例中。特别地，提供了Ag40和Ag45的衍生物(例如实施例中所示的衍生物18-10，它相对于Ag45具有的改进的性质)。因此，对于B表位，它可以具有至少约4个氨基酸的长度；对于顺序T表位，它可以具有至少约8个氨基酸残基(特别是约8至约19个氨基酸残基)的长度。在一个具体实施方式中，慢病毒载体的重组多核苷酸编码疟疾寄生虫抗原的片段，特别是由全长(即天然)抗原的连续氨基酸残基的缺失产生的片段，前提是它保持天然抗原的引发寄主中免疫应答的能力。如上所定义的多肽衍生物应当被认为是本发明任何的实施方式或定义中所述抗原多肽的替代物，除非它在本公开的上下文中看起来不相关。
表述“T表位”和“B表位”分别指T细胞驱动的适应性免疫应答和B细胞驱动的免疫应答中涉及的抗原决定簇。特别地，当在合适的条件下递送至寄主时，所述T表位和B表位分别引发T细胞和B细胞免疫应答。根据一个具体实施方式，本发明的目标抗原多肽和这些抗原多肽的多肽衍生物包含介导CD8+T细胞应答的表位。在一个具体的实施方式中，可选地或累积地，本发明的抗原多肽和这些抗原多肽的多肽衍生物包含介导抗体应答的表位。
在本发明的一个具体实施方式中，该化合物的组合物包含至少一种(优选至少两种)抗原多肽，或当作为多核苷酸或作为重组载体(特别是慢病毒载体)提供时，化合物的组合物包含至少一种(优选至少两种)编码抗原多肽的重组多核苷酸；其中，所述抗原多肽为疟原虫寄生虫的环子孢子蛋白(CSP)或包括疟原虫寄生虫的环子孢子蛋白(CSP)，所述疟原虫寄生虫选自恶性疟原虫、三日疟原虫(Plasmodium malariae)、间日疟原虫、卵形疟原虫(Plasmodium ovale)或诺氏疟原虫和伯氏疟原虫，特别是选自恶性疟原虫和间日疟原虫。特别地，它是CSP的截短形式，特别是缺乏CSP的GPI锚定基序(GPI anchoring motif)的多肽。在此类化合物的组合物中，病毒(特别是慢病毒)载体中包含的其它多肽或多核苷酸也选自上述公开的疟原虫寄生虫。
在本发明的化合物的组合物的一个具体实施方式中，活性成分作为多核苷酸或作为表达抗原多肽的载体(特别是慢病毒载体)提供，其中，载体在它们的基因组(载体基因组)中具有或包含重组多核苷酸，该重组多核苷酸编码至少一种如实施例所示的伯氏疟原虫的CSP的多肽，或有利地，该重组多核苷酸编码如本文公开或说明的恶性疟原虫或间日疟原虫的直系同源序列，例如，对应于缺乏GPI锚定基序的所述CSP抗原片段的多肽(CSP的多肽衍生物)。所述GPI基序对应于CSP抗原的天然氨基酸序列中C末端部分的最后12个氨基酸残基。
在本发明的一个具体实施方式中，该化合物的组合物包含单独(separate)的活性成分或单一(single)或多种活性成分的单独组合物，或由单独的活性成分或单一或多种活性成分的单独组合物组成。这些活性成分作为单独组合物或包装组合提供，可用于分别施用给寄主或相反地联合施用给寄主。
在本发明的另一个具体实施方式中，该化合物的组合物包含所有活性成分的混合物或由所述活性成分的混合物组成，另有说明的情况下，该化合物的组合物由所述活性成分的单一组合物组成。
因此，特别地，本文公开的抗原多肽可以作为一组(collection)载体(特别是一组慢病毒载体，特别是一组基于HIV-1的载体)的表达产物提供，其中，每个载体表达一种以上抗原多肽，该组载体表达所有抗原多肽。该组载体可以作为施用的单一组合物提供，或作为同时或分别及时施用给寄主的单独组合物提供。
已证明，当作为组合一起使用时，这些抗原多肽(特别是作为表达该抗原多肽的载体提供时)适用于引发保护性免疫应答，特别是在用疟原虫寄生虫(所述多肽来源于该疟原虫寄生虫)免疫的非人哺乳动物中针对严重攻击的无菌保护。因此，通过设计可用于制备候选疫苗的活性成分，本发明的化合物的组合物提供了对于针对人类寄主中疟原虫感染的有效替代需求的回应。
无论它作为一种或多种组合物呈现，本发明的化合物的组合物提供单个的和集合的活性成分(作为抗原多肽或作为载体颗粒，特别是慢病毒载体颗粒)，这些活性成分共同构成一剂候选药物产品的定性组成。
在本发明的化合物的组合物的一个具体实施方式中，活性成分由人感染(human-infecting)疟原虫寄生虫的抗原多肽组成，或活性成分由表达人感染疟原虫寄生虫的抗原多肽的慢病毒载体组成，或活性成分包含此类抗原多肽和病毒载体(特别是慢病毒载体)的混合物或结合物，特别地，其中，疟原虫寄生虫是恶性疟原虫或间日疟原虫。
在本发明的特定化合物的组合物的一个具体实施方式中，环子孢子蛋白(CSP)是该蛋白的全球分布的变体的代表，例如CSP VK210(参考GenBank：AAKM01000017.1和蛋白质IDXP_001613068)或CSP VK247(参考GenBank：GU339076.1和蛋白质ID：ADB92545.1)。
本发明的特定化合物的组合物的特征在于活性成分包含以下抗原或由以下抗原组成：序列为SEQ ID NO：11、12、14、15、17或18的环子孢子蛋白(CSP)；序列为SEQ ID NO：20、21、23、24、26或27的血小板反应蛋白相关匿名蛋白(TRAP)；序列为SEQ ID NO：29、30、32、33、35或36的半胱氨酸蛋白酶抑制剂(ICP)；序列为SEQ ID NO：38、39、41、42、44、45或47的金属肽酶(Bergheilysin/Falcilysin)；序列为SEQ ID NO：58、59、61、62、64或65的GPI锚定蛋白P113；序列为SEQ ID NO：49、50、52、53、55或56的成孔样蛋白SPECT2；序列为SEQ IDNO：67、68、70、71、73或74的蛋白Ag40或它的变体；以及序列为SEQ ID NO.76、77、79、80、82或83的蛋白Ag45(11-10)或它的变体；或它们的多肽衍生物，该多肽衍生物由与衍生其的(恶性疟原虫或间日疟原虫)抗原多肽具有至少86％的氨基酸同一性(优选至少95％的氨基酸同一性)的氨基酸序列组成；并且，当该多肽衍生物被包含在本发明的组合化合物中时，它具有衍生其的抗原多肽的保护性。86％的氨基酸同一性阈值对应于：比较称为3D7菌株的参考菌株(它的相关抗原的氨基酸序列为本文提供的那些)的8种恶性疟原虫红前期抗原与上文所述Genbank数据库中的其他恶性疟原虫寄生虫的序列时，获得的三种最不相似的Pf保护性抗原(PfCSP；查询覆盖率为100％，氨基酸同一性为86％)的平均同一性。
特别地，通过疟原虫抗原的原始序列中的氨基酸残基置换，和/或通过所述原始序列中短序列的点突变(例如置换、插入或缺失)或缺失，获得如此公开的特定多肽衍生物；前提是所衍生多肽基本上保持他所来源多肽的免疫原性特性。因此，衍生物可以通过多肽来说明，在所述多肽序列中包含残基，所述残基来源于获得该衍生物的多核苷酸构建体(construct)，例如由多核苷酸中存在的Kozak序列而得到的氨基酸残基。其他衍生物可以通过抗原序列的原始氨基酸残基的氨基酸残基保守置换获得，特别是小于20％的氨基酸置换，特别是小于15％或小于5％的氨基酸置换，特别是小于3％或小于2％的氨基酸置换。不考虑可选地添加功能性氨基酸序列至抗原多肽的天然或突变ORF(开放阅读框)，特别地，通过置换(特别是氨基酸残基的保守置换)获得的此类衍生物具有与它们所来源的原始序列相同的长度。或者，当衍生多肽具有ORF(相对于原始ORF，所述ORF包含通过缺失或添加得到的突变体)时，关于所表达的多肽衍生物的氨基酸残基数目，所测定的突变ORF的长度有利地为原始序列长度的至少95％，优选与原始序列至少97％、98％或99％相同。
在化合物的组合物的一个具体实施方式中，其中，活性成分包含人慢病毒载体(特别是HIV-1慢病毒载体)或由人慢病毒载体(特别是HIV-1慢病毒载体)组成，该人慢病毒载体表达抗原多肽或其多肽衍生物，其中，所述抗原多肽或其多肽衍生物：
-由单独的载体单个地表达，和/或
-由一种以上载体表达，其中，至少一种载体表达超过一种的抗原多肽或它的多肽衍生物。
在此种化合物的组合物的一个具体实施方式中，其中，活性成分是慢病毒载体，特别是基于HIV-1的载体，每种慢病毒载体是复制缺陷的(replication-incompetent)假型(pseudotyped)慢病毒载体，特别是复制缺陷的假型HIV-1慢病毒载体，其中，所述载体含有基因组，所述基因组包含哺乳动物密码子优化的合成核酸，特别是人类密码子优化的合成核酸，其中，所述合成核酸编码感染哺乳动物(特别是人类寄主)的疟原虫寄生虫的抗原多肽或它(们)的多肽衍生物。特别地，疟疾寄生虫可以是恶性疟原虫、间日疟原虫、诺氏疟原虫、食蟹猴疟原虫、三日疟原虫、卵形疟原虫。
特别地，使用载体颗粒基因组中密码子优化的序列，使得抗原多肽可以在施用载体的寄主细胞中强烈表达，特别是通过改善mRNA稳定性或减少二级结构。此外，表达的抗原多肽经历翻译后修饰，翻译后修饰适用于寄主细胞中的抗原多肽的加工，特别是通过修饰编码的多肽中的翻译修饰位点(例如糖基化位点)。密码子优化工具在本领域中是公知的，包括算法和服务，例如由GeneArt(Life technologies-USA)和DNA2.0(Menlo Park，California-USA)提供的算法和服务。在一个具体实施方式中，对编码抗原多肽或它的衍生物的ORF序列进行密码子优化，并且优化在将编码ORF的序列导入质粒制备载体基因组之前进行。在另一个实施方式中，载体基因组的其他序列也是密码子优化的。
由病毒载体组成的活性成分可以是整合(integrative)假型化的慢病毒载体，特别是复制缺陷的整合假型化的慢病毒载体，特别是HIV-1载体。此类慢病毒载体可另外含有基因组，所述基因组包含哺乳动物密码子优化的合成核酸，特别是人类密码子优化的合成核酸，其中，所述合成核酸编码如本文所公开的感染哺乳动物的疟原虫寄生虫(特别是如本文公开的感染人类寄主的寄生虫)的抗原多肽或它的多肽衍生物。
或者，慢病毒载体(特别是基于HIV-1的载体)可以是非整合(non-integrative)复制缺陷的假型慢病毒载体。
适用于实现本发明的慢病毒载体的具体实施方式涉及一种慢病毒载体，该慢病毒载体的基因组获自pTRIP载体质粒，其中，在哺乳动物细胞的功能性启动子(特别是人β2微球蛋白启动子)的控制下，可选地，在旱獭肝炎病毒(WPRE，woodchuck hepatitis virus)的转录后调控元件(post-transcriptional regulatory element)的控制下，克隆了编码抗原多肽或其多肽衍生物的疟原虫合成核酸。
在本发明的另一个实施方式中，表达本文所述特征的抗原多肽的慢病毒载体用来自印第安纳(Indiana)血清型或新泽西(New-Jersey)血清型水疱性口炎病毒(V-SVG，Vesicular Stomatitis Virus)糖蛋白G进行假型化。
这些慢病毒载体的具体特征将在下文进一步详细讨论。
本发明的化合物的组合物中所包含的抗原多肽可以有利地由核酸分子表达，特别是由哺乳动物密码子优化的合成核酸表达，所述核酸分子的特征在于具有以下序列：
-伯氏疟原虫的CSP的SEQ ID NO：10、伯氏疟原虫的TRAP的SEQ ID NO：19、伯氏疟原虫的ICP的SEQ ID NO：28、伯氏疟原虫的Falcilysin的SEQ ID NO：37、伯氏疟原虫的GPI锚定蛋白P113的SEQ ID NO：57、伯氏疟原虫的成孔样蛋白SPECT2的SEQ ID NO：48、伯氏疟原虫的蛋白Ag40 119-09的SEQ ID NO：66和伯氏疟原虫的蛋白Ag45 11-10的SEQ ID NO：75，或者，
-恶性疟原虫的CSP的SEQ ID NO：13、恶性疟原虫的TRAP的SEQ ID NO：22、恶性疟原虫的ICP的SEQ ID NO：31、恶性疟原虫的Falcilysin的SEQ ID NO：40、恶性疟原虫的成孔样蛋白SPECT2的SEQ ID NO：51、恶性疟原虫的GPI锚定蛋白P113的SEQ ID NO：60、恶性疟原虫的蛋白11-09的SEQ ID NO：69和恶性疟原虫的蛋白11-10的SEQ ID NO：78，或者，
-间日疟原虫的CSP的SEQ ID NO：16、间日疟原虫的TRAP的SEQ ID NO：25、间日疟原虫的ICP的SEQ ID NO：34、间日疟原虫的Falcilysin的SEQ ID NO：43、间日疟原虫的成孔样蛋白SPECT2的SEQ ID NO：54、间日疟原虫的GPI锚定蛋白P113的SEQ ID NO：63、间日疟原虫的蛋白11-09的SEQ ID NO：72和间日疟原虫的蛋白11-10的SEQ ID NO：81。
多核苷酸的密码子优化可影响编码的多肽的翻译后修饰(特别是当它在哺乳动物细胞中表达时)，因此使得能表达具有结构特征的多肽，所述结构特征区别于由相应的非优化序列编码的多肽的结构特征(Mauro V.P.和Chappell S.A.，Trends Mol Med，2014年11月；20(11)：604-613)。
类似地，化合物的组合物中表达抗原多肽的慢病毒载体可以有利地在它们的基因组中含有核酸分子，所述核酸分子是哺乳动物密码子优化的合成核酸，所述核酸分子的特征在于具有以下序列：
-伯氏疟原虫的CSP的SEQ ID NO：10、伯氏疟原虫的TRAP的SEQ ID NO：19、伯氏疟原虫的ICP的SEQ ID NO：28、伯氏疟原虫的Falcilysin的SEQ ID NO：37、伯氏疟原虫的GPI锚定蛋白P113的SEQ ID NO：57、伯氏疟原虫的成孔样蛋白SPECT2的SEQ ID NO：48、伯氏疟原虫的蛋白Ag40 19-09的SEQ ID NO：66和伯氏疟原虫的蛋白Ag45 11-10的SEQ ID NO：75，或者
-恶性疟原虫的CSP的SEQ ID NO：13、恶性疟原虫的TRAP的SEQ ID NO：22、恶性疟原虫的ICP的SEQ ID NO：31、恶性疟原虫的Falcilysin的SEQ ID NO：40、恶性疟原虫的成孔样蛋白SPECT2的SEQ ID NO：51、恶性疟原虫的GPI锚定蛋白P113的SEQ ID NO：60、恶性疟原虫的蛋白11-09的SEQ ID NO：69和恶性疟原虫的蛋白11-10的SEQ ID NO：78，或者
-间日疟原虫的CSP的SEQ ID NO：16、间日疟原虫的TRAP的SEQ ID NO：25、间日疟原虫的ICP的SEQ ID NO：34、间日疟原虫的Falcilysin的SEQ ID NO：43、间日疟原虫的成孔样蛋白SPECT2的SEQ ID NO：54、间日疟原虫的GPI锚定蛋白P113的SEQ ID NO：63、间日疟原虫的蛋白11-09的SEQ ID NO：72和间日疟原虫的蛋白11-10的SEQ ID NO：81。
本发明还涉及适合给哺乳动物寄主施用的制剂，该制剂包含本文提供的任一定义的化合物的组合物作为活性成分，所述活性成分用于保护免受疟原虫感染或寄生虫诱导的病症或疾病，并且该制剂包含适用于给有需要的寄主(特别是人类寄主)施用的赋形剂。
在本发明的另一方面，本发明的化合物的组合物或包含该化合物的组合物的制剂用于在哺乳动物寄主(特别是人类寄主)中引发针对疟疾寄生虫感染或针对寄生虫诱导的病症或疾病的保护性免疫；可选地，与适当的递送载体和可选的佐剂组分和/或免疫刺激剂组分结合。
因此，当本发明的化合物的组合物(特别是慢病毒载体颗粒)施用于有需要的寄主(特别是哺乳动物，特别是人类寄主)时引发免疫应答，包括激活幼稚淋巴细胞，以及针对疟疾寄生虫抗原产生效应T细胞应答和产生抗原特异性的记忆T细胞免疫应答。另外地，在施用化合物的组合物后，免疫应答可能涉及针对递送至寄主或在寄主中表达的抗原多肽的体液应答。免疫应答可以阻止当此种寄生虫以子孢子形式接种到寄主时的疟疾寄生虫感染，或者可以防止由子孢子形式的疟疾寄生虫接种引起的病理状态的发作(onset)或发展，或者防止所述寄生虫进一步形式(如裂殖子形式)产生的后果的发作或发展。
因此，本发明的化合物的组合物的活性成分适用于引发针对寄生虫感染或针对寄生虫诱导的疾病或病症的保护性免疫应答。特别地，此种应答能够控制或抑制由寄生虫侵入或疟疾寄生虫周期中的外红期(即肝期)诱导所引起的病理发作；在一个有利的实施方式中，此应答适用于预防、减轻或抑制所述寄生虫的红内期(erythrocytic stage)的发作或发展。有利地，已观察到，特别是当活性成分作为单一施用方案或“引发-加强免疫(prime-boost)”施用方案中使用的慢病毒载体颗粒提供时，本发明的化合物的组合物能够产生保护性免疫，特别是能够产生针对疟疾寄生虫诱导的病状的无菌保护。可以通过控制寄生虫周期中的肝期(如果不是之前)感染的后果而获得此种无菌保护。
在本发明的一个具体实施方式中，特别是当活性成分作为慢病毒载体提供时，该化合物的组合物是用适于施用的载体配制的悬浮液，用于给寄主施用。可以根据免疫组合物的施用途径选择生理学上可接受的载体。在一个优选的实施方式中，可以通过注射(特别是肌内)进行施用，或者对于儿童，可以鼻内施用或局部皮肤涂敷。
本发明的化合物的组合物用于哺乳动物寄主(特别是在人类寄主)中针对疟疾寄生虫感染或针对寄生虫诱导的疾病或病症的保护性免疫，所述用途涉及免疫模式，该免疫模式包括：施用有效量的活性成分，引发寄主的细胞免疫应答和/或体液免疫应答，可能作为引发，并在适当的时候及时施用有效量的所述活性成分以加强寄主的细胞免疫应答，并可选地重复(一次以上)所述施用步骤以加强免疫，其中，如果活性成分作为慢病毒颗粒提供而在各引发或加强免疫步骤中施用，则用不相互交叉中和(cross-neutralise)的不同假型包膜蛋白对它们进行假型化，并且其中，所述引发和加强免疫步骤在时间上隔开至少6周，特别是至少8周。
有关施用方案的详细信息将在下文进一步讨论。
特别地，本发明的化合物的组合物作为慢病毒载体被特别用于特定实施方式中，用于哺乳动物寄主(特别是人类寄主)中针对疟疾寄生虫感染或抗寄生虫诱导的病状的保护性免疫，以至少获得细胞免疫应答(T细胞免疫应答)，特别是CD8介导的细胞免疫应答或CD4介导的细胞免疫应答，即由携带CD8或CD4受体的活化细胞介导的免疫应答；当定义本发明的慢病毒颗粒的免疫方案时，优选有利地靶向细胞毒性T淋巴细胞(CTL)和记忆T细胞应答。
免疫应答还可以涉及由所述化合物引发的针对所述至少一种抗原多肽产生的体液应答(即抗体)。在一个具体实施方式中，所述体液应答是保护性体液应答。保护性体液应答主要产生对它们的抗原具有高亲和力的成熟抗体(例如IgG或IgM)。在一个具体方面，保护性体液应答是T细胞依赖性的。在一个具体实施方式中，保护性体液应答诱导中和抗体的产生。
在本发明的一个具体实施方式中，特别是当活性成分是慢病毒载体时，即使以具有缺陷型整合酶的形式使用本发明的化合物的组合物，本发明的化合物的组合物也能够引发早期免疫应答。表述“早期免疫应答”指在施用产品后约一周内获得的保护性免疫应答(针对寄生虫或针对寄生虫诱导的病状的保护)。
在另一个特别有利的实施方式中，通过本发明的化合物的组合物(特别是作为慢病毒颗粒)获得的免疫应答是持久的免疫应答，即所述免疫应答包括记忆细胞应答，特别是中枢记忆细胞应答；在一个具体实施方式中，至少几个月后，应答仍然可以被检测到。
当免疫应答包括体液应答时，可以通过任何合适的方法(如ELISA、免疫荧光(IFA)、调焦中和试验(FRNT，focus reduction neutralization test)、免疫沉淀或Western Blotting))检测特异性抗体，来显示持久的应答。
根据使用本发明的化合物的组合物的一个具体方面，活性成分被设计成能够在有需要的寄主中进行引发-加强免疫施用，其中第一施用步骤引发免疫应答(特别是细胞免疫应答)，后续施用步骤加强免疫反应(包括细胞免疫应答)。对于每个施用步骤，优选载体颗粒的假型包膜蛋白与其他步骤中使用的那些不同，特别是源自不同病毒(特别是不同VSV)。在引发-加强免疫的方案中，每个步骤施用的化合物的组合物包含如本文所定义的慢病毒载体，该慢病毒载体共同表达所有抗原多肽。因此，可提供化合物的组合物以进行引发-加强免疫方案，该化合物的组合物包含化合物，所述化合物至少由于它们的假型包膜蛋白的差异而为不同的慢病毒颗粒。
因此，当选择引发-加强免疫方案时，含有所述慢病毒载体的化合物的组合物可以在单独的包装中提供，或者可以在共同的包装中提供用于单独使用。
因此，包含在包装中并包含使用说明的注意事项可以指示化合物的组合物的施用顺序和它们施用的时间顺序，用于在寄主中进行引发并随后加强免疫应答。
根据本发明，当化合物的组合物用于引发-加强免疫方案时，提供：第一化合物的组合物，该第一化合物的组合物含有慢病毒载体颗粒，该慢病毒载体颗粒用从VSV(新泽西株)获得的第一种确定的假型包膜G蛋白进行假型化；第二种化合物的组合物，该第二种化合物的组合物含有慢病毒病毒载体颗粒，该慢病毒载体颗粒用从VSV(印第安纳株)获得的第二种确定的假型包膜G蛋白进行假型化。可选地，可以颠倒引发-加强免疫方案中如此描述的第一和第二组合的使用顺序。因此，当用于引发-加强免疫方案时，至少由于用于假型化载体颗粒的特定假型包膜蛋白，包含在本发明的单独化合物的组合物中的慢病毒载体颗粒彼此不同。
在下文的实施例中，已根据引发-加强免疫方案用本发明的慢病毒载体颗粒对小鼠模型进行治疗，发明人已表明：在根据此种方案免疫的并在最后一次免疫步骤后受到攻击的小鼠中，大部分接种疫苗的小鼠(超过80％)表现出无菌保护，这说明本发明的化合物的组合物在寄主中引发有效保护，因此构成用于免疫(特别是人类寄主)的合适候选疫苗。
在一个具体实施方式中，本发明涉及本发明的化合物的组合物，特别是如本文所定义的慢病毒载体颗粒，用于按照剂量方案在哺乳动物寄主(特别是在人类寄主)中引发针对疟疾寄生虫感染或针对寄生虫诱导的病状的保护性免疫，在剂量方案(包含单独提供的所述化合物的组合物的活性成分)中，其中，用于引发和加强细胞免疫应答的活性成分的剂量是中等剂量，并且用于加强细胞免疫应答的剂量高于引发剂量。
因此，在施用模式中使用的用于引发和加强细胞免疫应答的慢病毒载体的剂量包括：当使用整合型载体时，每种类型的病毒颗粒105TU至1010TU，特别是从105至107。当使用整合缺陷型(integrative-incompetent)载体时，用于引发和加强免疫的剂量包括：每种类型的慢病毒颗粒107至1010。
本发明还涉及本发明的化合物的组合物(特别是本文所定义的慢病毒载体)的用途，用于制备哺乳动物寄主中(特别是在人类寄主中)针对疟疾寄生虫感染或针对寄生虫诱导的病状的预防性免疫的免疫原性组合物。
根据本公开，本发明还涉及在哺乳动物寄主中(特别是在人类寄主中)提供免疫的方法，该方法包括以下步骤：施用本发明的化合物的组合物(特别是本发明的慢病毒载体)，引发免疫应答；以及可选地，将施用步骤重复一次或多次，加强所述应答。
在本发明的一个具体实施方式中，化合物的组合物(特别是作为本发明的慢病毒载体提供)可以与适合施用于哺乳动物(特别是人类寄主)的佐剂化合物和/或免疫刺激化合物以及适当的递送载体联合使用。
上述化合物的组合物可以通过不同途径注射到寄主中：皮下(s.c.)、皮内(i.d.)、肌内(i.m.)或静脉内(i.v.)注射、口服施用和粘膜或皮肤施用，特别是鼻内施用或吸入。施用量(剂量)取决于待治疗的受试者，包括考虑患者的状况、个体免疫系统的状态、施用途径和寄主的大小。可以确定相对于载体颗粒(对于HIV-1慢病毒载体)的等效转导单位中含量的表达的合适剂量范围。
当阅读说明慢病毒载体颗粒的制备和应用的实施例和附图时，本发明的其他实施例和特征将是显而易见的，所述慢病毒载体颗粒具有可以与本说明书中给出的定义单独组合的特征。
根据本发明使用的慢病毒载体的详细描述
因此，本发明涉及慢病毒载体，它是慢病毒颗粒(即载体颗粒)，并且可以是复制缺陷的慢病毒载体，特别是基于复制缺陷的HIV-1的载体，其特征在于：(i)它们用确定的异源病毒包膜蛋白或来自非HIV的RNA病毒的病毒包膜蛋白进行假型化，并且(ii)它们在其基因组中包含编码至少一种抗原多肽(或其多肽衍生物)的至少一种重组多核苷酸，其中，所述抗原多肽携带疟原虫寄生虫的红前期抗原的表位或其多肽衍生物，其中，所述寄生虫能够感染哺乳动物寄主，并且，所述表位包括T表位。
在本发明的一个具体实施方式中，所编码的疟原虫寄生虫的红前期抗原的抗原多肽还包含B表位。
由载体表达的抗原多肽或它们的衍生物是本发明的任何方面中本文公开的那些，特别是本发明的化合物的组合物的描述中的那些。
根据本发明的一个具体实施方式，慢病毒载体或者被设计成表达成熟(即整合型)或缺陷(即整合缺陷型)的颗粒。
慢病毒载体的制备是本领域技术人员公知的，并且已在文献中广泛公开(参见综述Sakuma T.等的，Biochem.J.(2012)443，603-618)。本文还在实施例中说明了这些载体的制备。
在本发明的一个具体实施方式中，编码慢病毒载体的抗原多肽(ORF)的多核苷酸是经哺乳动物密码子优化的(CO，codon optimized)，特别是经人类密码子优化的。可选地，所述颗粒的基因组的慢病毒序列也具有哺乳动物密码子优化的核苷酸序列。在本发明的一个特定方面，小鼠细胞中的表达已进行密码子优化。在另一个实施方式中，编码慢病毒载体的抗原多肽的多核苷酸的序列是经人类密码子优化(CO)的。
已观察到：密码子优化的核苷酸序列，特别是当针对哺乳动物(特别是人细胞)中的表达进行优化时，使得能够在此类哺乳动物或人细胞中产生更高产率的颗粒。实施例中说明了细胞制备。因此，当将本发明的慢病毒载体颗粒施用于哺乳动物(特别是人类寄主)时，在所述寄主中产生更大量的颗粒，这有利于引发强烈的免疫应答。
本发明中定义的慢病毒载体(即，慢病毒载体颗粒或基于慢病毒的载体颗粒)是由携带包膜蛋白的载体颗粒组成的假型化的慢病毒载体，所述包膜蛋白源自不同于特定慢病毒的病毒(特别是不同于HIV的病毒，特别是不同于HIV-1的病毒)，这提供了慢病毒载体颗粒的载体基因组。因此，相对于颗粒的载体基因组，所述包膜蛋白是“异源的”病毒包膜蛋白或病毒包膜蛋白。在下文中，提及“包膜蛋白”时，还包括适合于实施本发明的任何类型的包膜蛋白。
当在本申请中提及“慢病毒”载体(基于慢病毒的载体)时，它特别涉及基于HIV的载体，特别是基于HIV-1的载体。
适合于实施本发明的慢病毒载体是所谓的置换型载体(replacement vector)，意指编码慢病毒蛋白的原始慢病毒序列在载体的基因组中基本上缺失；或者当存在时，它被修饰(特别是被突变，特别是被截短)，以阻止生物学活性的慢病毒蛋白的表达，特别地，在HIV的情况下，以阻止所述转移载体表达功能性ENV、GAG和POL蛋白以及可选的慢病毒(特别是HIV)的其他结构蛋白和/或辅助蛋白和/或调节蛋白。在一个具体实施方式中，慢病毒载体是第一代载体，特别是第一代基于HIV的载体，其特征在于它是使用单独的质粒获得的，以提供：(i)包装构建体、(ii)包膜和(iii)转移载体基因组。或者，它可以是第二代载体，特别是第二代基于HIV的载体，此外，它缺乏病毒辅助蛋白(例如在HIV-1、Vif、Vpu、Vpr或Nef的情况下)，因而它仅包括九个HIV全基因中的四个：gag、pol、tat和rev。在另一个实施方式中，载体是第三代载体，特别是第三代基于HIV的载体，它进一步缺乏所述病毒辅助蛋白并且也是Tat非依赖性的；这些第三代载体可使用提供载体的功能元件的4种质粒获得，包括编码HIV的Rev蛋白的一种质粒(当载体基于HIV-1时)。该载体系统仅包含HIV-1的9种基因中的3种。这些基于HIV的载体的结构和设计在本领域中是公知的。
载体颗粒的“载体基因组”是还包含作为重组序列的目的多核苷酸或转基因的重组核酸，它编码如本文所公开的疟疾寄生虫的一种以上抗原多肽或它(们)的多肽衍生物。载体基因组的基于慢病毒的序列和多核苷酸/转基因由质粒载体携带，从而产生“转移载体”，也称为“序列载体”。因此，这些表达在本说明书中可互换使用。根据一个具体实施方式，本发明制备的载体基因组包含核酸，该核酸具有选自SEQ ID NO：10、13、16、19、22、25、28、31、34、37、40、43、46、48、51、54、57、60、63、66、69、72、75、78、81的序列或包含多个编码抗原多肽或它们的衍生物的这些序列。
因此，除了包含编码疟疾寄生虫(在它表达的适当调节序列的控制下)的抗原多肽或它们的多肽衍生物的所谓重组多核苷酸外，如本文所定义的载体基因组还包含原始慢病毒基因组的序列，所述原始慢病毒基因组的序列为所述基因组的非编码区并且是提供DNA或RNA合成和加工的识别信号所必需的(小病毒基因组)。这些序列是顺式作用序列(cis-acting sequence)，它们对于包装(ψ)、逆转录(可能相对于原始序列突变的LTR)和转录和可选的整合(RRE)所必需的；为了本发明的特定目的，它们含有功能性序列，在所述细胞中，该功能性序列有利于细胞中的核转运从而有利于转基因转移效率，该元件被描述为DNAFlap(“翻卷”)元件，Flap元件包含或由慢病毒基因组序列(特别是在HIV-1中，或在逆转录因子一些中，如酵母的那些)中存在的所谓的中心cPPT-CTS核苷酸结构域组成。
基于本领域所述原理和主要公开于(Zennou等，2000；Firat H.等，2002；VandenDriessche T.等)的此种慢病毒载体的实例，使用载体基因组的结构和组成制备本发明的慢病毒载体。将如本文所述的此种类型的构建体已经存放在CNCM(Institut Pasteur，France)。在此方面，本公开也参考了专利申请WO99/55892、WO01/27300和WO01/27304，包括保藏的生物材料。
根据本发明的一个具体实施方式，载体基因组可以是置换型载体，在该置换型载体中，2个长末端重复序列(LTR)之间的所有病毒蛋白编码序列已经被编码疟原虫多肽的重组多核苷酸置换，其中，DNA-Flap元件已被重新插入相关的本文所述的所需顺式作用序列。与颗粒的制备相关的载体基因组的组成的其他特征是公开的。
在本发明的一个具体实施方式中，一种慢病毒载体编码疟原虫寄生虫的一种抗原多肽。
在一个具体实施方式中，本发明的慢病毒载体可在其基因组中包含一种以上重组多核苷酸，该重组多核苷酸编码至少一种抗原多肽，该抗原多肽携带本文公开的红前期抗原的表位。特别地，所述载体基因组包含在基因组中连续或分开的两个多核苷酸，该多核苷酸编码疟原虫寄生虫的相同或不同红前期抗原的不同多肽或寄生虫的不同抗原的不同抗原多肽衍生物。
在一个具体实施方式中，载体基因组含有两个或多个重组多核苷酸，每个重组多核苷酸编码不同的抗原多肽，每个多肽来源于本文公开的不同红前期抗原，包括CSP抗原和至少一种Ag40或Ag45抗原，以及可选的一种以上抗原，该抗原包含选自下组的抗原多肽：本文所公开疟原虫寄生虫的血小板反应蛋白相关匿名蛋白(TRAP)、半胱氨酸蛋白酶抑制剂(ICP)、金属肽酶(Falcilysin)、GPI锚定蛋白P113、成孔样蛋白SPECT2或它们的衍生物。
本文关于抗原多肽的描述类似地适用于它们的多肽衍生物。
实施例中还公开了本发明各种实施方式使用的慢病毒载体的特定特征，此类特征可以单独使用或组合使用以产生载体。
根据本发明，慢病毒载体颗粒用异源病毒包膜蛋白(viral envelope protein)或来源于RNA病毒的包膜的病毒多聚蛋白(polyprotein)假型化，该RNA病毒不是提供慢病毒颗粒的慢病毒基因组序列的慢病毒。
作为制备慢病毒载体的包膜蛋白的分型的实施例，本发明涉及水疱性口炎病毒(VSV)的病毒跨膜糖基化(所谓的G蛋白)包膜蛋白，例如，该蛋白选自印第安纳株的VSV-G蛋白和新泽西株的VSV-G蛋白。
水疱性口炎病毒的包膜糖蛋白(VSV-G)是用作野生型病毒颗粒的表面衣壳的跨膜蛋白。它也是用于基因工程慢病毒载体的合适衣壳蛋白。目前，9种病毒被明确分类在VSV属中，19种弹状病毒(rhabdovirus)被临时分类在该属中，所有病毒均显示不同程度的交叉中和(cross-neutralisation)。测序时，蛋白G基因显示出序列相似性。VSV-G蛋白包括N-末端胞外域、跨膜区和C-末端细胞质尾。它通过分泌面高尔基体(transGolgi)网络(内质网和高尔基体)运输到细胞表面。
水疱性口炎印第安纳型病毒(VSIV)和水疱性口炎新泽西型病毒(VSNJV)是假型化本发明的慢病毒载体或设计重组包膜蛋白以假型化该慢病毒载体的优选菌株。它们的VSV-G蛋白公开于GenBank中，其中呈现了几种菌株。对于新泽西株的VSV-G，特别参考登录号为V01214的序列。对于印第安纳株的VSV-G，参考Genbank中登录号为AAA48370.1的序列，其对应于菌株JO2428。
所述病毒包膜蛋白能够被抗原呈递细胞摄入，特别是通过融合和/或胞吞作用被树突细胞(包括肝脏树突细胞)摄入。在一个具体实施方式中，摄入效率可以用作选择进行假型化的VSV包膜的特征。在此方面，可考虑检测转导(transduction)的相对滴度(DC滴度/其他被转导细胞的滴度，例如293T细胞)，优选与DC融合的能力相对良好的包膜。
抗原呈递细胞(APC)特别是树突细胞(DC)是假型慢病毒载体的合适靶细胞，该慢病毒载体相应地用作免疫组合物。
VSV-G包膜蛋白由含有所述蛋白质的编码序列的多核苷酸表达，该多核苷酸被插入质粒(称为包膜表达质粒或假型化包膜质粒)，该质粒用于制备本发明的慢病毒载体颗粒。编码包膜蛋白的多核苷酸处于调控序列的控制下，以转录和/或表达编码序列(包括可选的转录后调控元件(PRE)，特别是诸如旱獭肝炎病毒的元件的多核苷酸(即WPRE序列)，可从Invitrogen商购)。
因此，本文提供了核酸构建体，该核酸构建体包含适用于哺乳动物细胞(特别是体内人细胞)的内部启动子(internal promoter)和核酸(在所述启动子控制下该核酸编码所述包膜蛋白)。含有该构建体的质粒被用于转染或转导适合于制备载体颗粒的细胞。特别地，启动子可以按它们的性质选择为组成型启动子、组织特异性启动子或诱导型启动子。合适启动子的实例包括以下基因的启动子：MHC I类基因启动子、人β-2微球蛋白基因(β2M启动子)、EF1α、人PGK、PPI(前胰岛素原)、硫代糊精(thiodextrin)、HLA DR恒定链(P33)、HLADRα链、铁蛋白L链或铁蛋白H链、凝乳酶β4、凝乳酶β10、胱抑素核糖体蛋白L41、CMVie或嵌合启动子如GAG(CMV早期增强子/鸡β肌动蛋白)，如Jones S.等(Jones S.等，Human GeneTherapy，20：630-640，2009年6月)所公开的。
这些启动子也可用于表达调控序列(regulatory expression sequence)和/或用于表达转移载体的抗原多肽，其中所述表达调控序列参与衣壳化质粒(encapsidationplasmid)的gag-pol衍生蛋白的表达。
或者，当包膜表达质粒用于在稳定包装细胞系(packaging cell line)中表达(特别是连续表达的病毒颗粒的稳定表达)时，表达包膜蛋白的内部启动子有利地是诱导型启动子，例如Cockrell A.S.等，Mol.Biotechnol.，(2007)36：184-204中公开的诱导型启动子。作为此类启动子的实例，参考四环素和蜕皮激素诱导型启动子。包装细胞系可以是STAR包装细胞系(参考Cockrell A.S.等(2007年)、Ikedia Y.等(2003年)，NatureBiotechnol.，21：569-572)或SODk包装细胞系，例如SODk0(包括SODk1和SODk3)衍生细胞系(参考Cockrell A.S.等(2007年)、CockrellA；Cockrell A.S.等(2006年)MolecularTherapy，14：276-284；Xu K.等(2001年)、Kafri T.等(1999年)，Journal of Virol.，73：576-584)。
根据本发明，慢病毒载体为从哺乳动物细胞回收的产物，所述哺乳动物细胞用下述共转染：
-载体质粒，该质粒包含：(i)慢病毒(特别是HIV-1)的包装、逆转录和转录所必需的顺式激活序列(cis-active sequence)，并且还包含功能性慢病毒(特别是源自HIV-1)的DNA flap元件；和(ii)多核苷酸，该多核苷酸在表达调控序列的控制下编码如本文所公开的疟疾寄生虫的一种以上抗原多肽(或它们的多肽衍生物)；并且可选地包含用于整合到寄主细胞基因组中的序列；
-表达质粒，该质粒编码来自RNA病毒的假型化包膜，所述表达质粒包含编码用于假型化的包膜蛋白的多核苷酸，其中，所述包膜假型化蛋白有利地来自VSV，特别地，所述包膜假型化蛋白是印第安纳株或新泽西株的VSV-G；以及
-衣壳化质粒，该质粒包含慢病毒(特别是HIV-1)的gag-pol包装序列(适用于产生整合型载体颗粒)或经修饰的gag-pol包装序列(适用于产生整合缺陷型载体颗粒)。
因此，本发明还涉及如上所述慢病毒载体颗粒，所述慢病毒载体颗粒是从以下质粒转染的稳定细胞系中回收的产物：
-载体质粒，该质粒包含：(i)慢病毒(特别是HIV-1)的包装、逆转录和转录所必需的顺式激活序列，还包含功能性慢病毒(特别是HIV-1)的DNA flap元件，可选地包含整合所必需的顺式激活序列，所述载体质粒还包含：(ii)鼠或人基因的密码子优化序列的多核苷酸，该多核苷酸在表达调控序列(特别是启动子)的控制下编码本文所公开疟原虫寄生虫的抗原多肽(或其衍生物)；
-VSV-G包膜表达质粒，该质粒包含编码VSV-G(特别是印第安纳株或新泽西株的VSV-G)包膜蛋白的多核苷酸，其中，所述多核苷酸受表达调控序列(特别是包含诱导型启动子的表达调控序列)的控制；以及
-衣壳化质粒，其中，该质粒包含慢病毒(特别是HIV-1)的gag-pol编码序列(适用于产生整合型载体颗粒)或经修饰的gag-pol编码序列(适用于产生整合缺陷型载体颗粒)；其中，所述gag-pol序列来自与DNA flap元件相同的慢病毒亚家族，其中，所述慢病毒gag-pol序列或经修饰的gag-pol序列受表达调控序列的控制。
特别地，表达本发明的载体颗粒的稳定细胞系通过质粒的转导获得。
根据本说明书中公开的任何实施方式，该多核苷酸编码疟疾寄生虫的至少一种抗原多肽。特别地，该多核苷酸编码这样的多肽，所述多肽是截短的经哺乳动物(特别是人)密码子优化的序列，其编码恶性疟原虫、间日疟原虫或伯氏疟原虫的此种抗原多肽。
在一个具体实施方式中，所述多核苷酸编码两种以上的疟疾寄生虫抗原多肽，该抗原多肽源自和/或来自各种实施方式中所公开的所述寄生虫的不同抗原。因此，载体质粒可以包含几种用于表达各种抗原多肽的表达盒(expression cassette)或者可以包含双顺反子或多顺反子表达盒，其中，编码各种多肽的多核苷酸由病毒来源的IRES序列(内部核糖体进入位点，Internal Ribosome Entry Site)间隔，或者它可以编码融合蛋白。
载体基因组包含的内部启动子控制多核苷酸的表达，该多核苷酸编码疟疾寄生虫的抗原多肽(作为转基因或在表达盒中)，所述内部启动子可以选自以下基因的启动子：人β-2微球蛋白基因(β2M启动子)、EF1α、人PGK、PPI(前胰岛素原)、硫代糊精、HLA DR恒定链(P33)、HLA DRα链、铁蛋白L链或铁蛋白H链、凝乳酶β4、凝乳酶β10、胱抑素核糖体蛋白L41、CMVie或嵌合启动子如GAG(CMV早期增强子/鸡β肌动蛋白)，公开于Jones S.等(2009年)。
还可以选择上述内部启动子中的启动子用于表达包膜蛋白和包装(gag-pol衍生的)蛋白。
或者，载体颗粒可以通过将本文公开的质粒共转染到稳定的包装细胞系(所述细胞系因此变得能够连续分泌载体颗粒)而产生。有利地，涉及包膜蛋白表达的表达调控序列中的启动子是诱导型启动子。
当制备基于人慢病毒(特别是基于HIV-1病毒)的慢病毒载体时，可以进行以下具体实施方式。
根据本发明，慢病毒载体的基因组衍生自人慢病毒(特别是HIV慢病毒)。特别地，假型慢病毒载体是基于HIV的载体，例如基于HIV-1或HIV-2的载体，特别是衍生自HIV-1M(例如BRU或LAI分离株)的载体。或者，提供载体基因组必需序列的慢病毒载体可以源自能够转导哺乳动物细胞的慢病毒，例如EIAV、CAEV、VISNA、FIV、BIV、SIV、HIV-2、HIV-O。
如上所述，当考虑除载体基因组最终包含的重组多核苷酸之外的载体基因组时，载体基因组是置换型载体，在该置换型载体中，原始慢病毒基因组中2个长末端重复序列(LTR)之间的核酸被限制为顺式作用序列，所述顺式作用序列用于DNA或RNA合成和加工，包括将转基因有效递送至寄主细胞核，或至少缺失或突变的基本核酸区段，该基本核酸区段能够表达慢病毒结构蛋白，包括生物功能性GAG多蛋白和可能的POL和ENV蛋白。
在一个具体实施方式中，慢病毒的5'LTR和3'LTR序列用于载体基因组，但相对于原始慢病毒的3'LTR，3'-LTR至少在U3区域被修饰，例如，它可以缺失或部分缺失增强子。5'LTR也可以被修饰，特别是在它的启动子区域，其中，例如可以用不依赖于Tat的启动子置换U3内源启动子。
在一个具体实施方式中，载体基因组包含(HIV-1慢病毒载体的)Vif-、Vpr、Vpu-和Nef-辅助基因的一种或几种编码序列。或者，这些序列可以彼此独立地或彼此缺失，或者可以是非功能性的(第二代慢病毒载体)。
慢病毒载体颗粒的载体基因组包含作为插入的顺式作用片段的至少一种多核苷酸，该多核苷酸由DNA flap元件组成或包含该DNA flap元件。在一个具体实施方式中，DNAflap被插入在编码疟原虫寄生虫抗原多肽的多核苷酸的上游，并且有利地(但不是必须)位于载体基因组中的近中心位置。适用于本发明的DNA flap可以从逆转录病毒(特别是慢病毒，特别是人慢病毒，特别是HIV-1逆转录病毒)，或从逆转录病毒样生物(例如逆转录转座子)获得。或者，它可以从CAEV(山羊关节炎脑炎病毒)病毒、EIAV(马传染性贫血病毒)病毒、VISNA病毒、SIV(猴免疫缺陷病毒)病毒或FIV(猫免疫缺陷病毒)病毒中获得。DNA flap可以合成(化学合成)制备，或可以通过扩增DNA(该DNA提供如上定义的适当来源的DNA Flap)来制备，例如通过聚合酶链式反应(PCR)。在更优选的实施方式中，DNA flap获自HIV逆转录病毒，例如HIV-1或HIV-2病毒，包括这两种类型的任何分离株。
DNA flap(也称为cPPT/CTS)(定义于Zennou V.等，ref 27，2000，Cell，第101卷，173-185或WO99/55892和WO01/27304中)是处于一些慢病毒(特别是HIV)基因组的中心的结构，它通常在HIV逆转录期间导致合成三链DNA结构，并且它充当HIV基因组入核转运(nuclear import)的顺式决定簇(cis-determinant)。逆转录期间，DNA flap能够通过中央多嘌呤管道(cPPT，central polypurine tract)和中央终止序列(CTS，centraltermination sequence)顺式控制中心链置换事件(central strand displacementevent)。当插入慢病毒衍生的载体中时，多核苷酸(能够在逆转录过程中产生DNA flap)刺激基因转移效率并补充入核转运水平至野生型水平(Zennou等，Cell，2000年，第101卷，173-185或WO99/55892和WO01/27304)。
DNA flap的序列已在现有技术中，特别是在上述引用的专利申请中公开。这些序列也公开于SEQ ID NO：1序列的第2056位至第2179位。在载体基因组中，它们优选作为片段插入，可选地具有另外的侧翼序列(flanking sequence)，优选位于所述载体基因组中心附近的位置。或者，它们可以直接插入启动子的上游，该启动子控制编码抗原多肽的多核苷酸的表达。插入载体基因组中的所述片段包含DNA flap，所述片段可具有约80至约200bp的序列，这取决于其来源和制备方式。
根据一个具体实施方式，DNA flap具有约90至约140个核苷酸的核苷酸序列。
在HIV-1中，DNA flap是稳定的99个核苷酸长的正链(plus strand)重叠(overlap)序列。当用于本发明的慢病毒载体的基因组载体中时，特别是当它作为PCR片段制备时，它可以以较长的序列插入。一个特定的合适多核苷酸(其包含提供DNA flap的结构)是包含HIV-1DNA的cPPT和CTS区的124碱基对聚合酶链反应(PCR)片段(如SEQ ID NO：1所公开)。
明确指出，基因组载体中使用的DNA flap和衣壳化质粒的多核苷酸(其编码GAG和POL多蛋白)应来源于相同的慢病毒亚家族或来自相同的逆转录病毒样生物。
优选地，基因组载体的其他顺式激活序列也来源于与提供DNA flap的相同的慢病毒或逆转录病毒样生物。
载体基因组可以进一步包含用于克隆重组多核苷酸的一个或几个独特的限制性位点。
在一个优选的实施方式中，在所述载体基因组中，慢病毒载体基因组的3'LTR序列至少缺乏活化剂(增强子)和可能缺乏U3区的启动子。在另一个具体实施方式中，3'LTR区域缺乏U3区(ΔU3)。在此方面，参考WO01/27300和WO01/27304中的描述。
在一个具体实施方式中，在载体基因组中，LTR 5'的U3区被非慢病毒U3或被适用于驱动tat非依赖性初级转录的启动子置换。在此种情况下，载体不依赖于tat反式激活因子(第三代载体)。
载体基因组还包含psi(ψ)包装信号。包装信号衍生自gag ORF的N-末端片段。在一个具体实施方式中，其序列可以通过移码突变进行修饰，以防止转基因的转录/翻译对gag肽的可能的转录/翻译的任何干扰。
可选地，载体基因组还可包含选自剪接供体位点(SD)、剪接受体位点(SA)和/或Rev反应元件(RRE)的元件。
根据一个具体实施方式，载体质粒(或添加的基因组载体)包含以下用于转基因表达盒的顺式作用序列：
1.逆转录所需的LTR序列(长末端重复)、转录所需的序列，和可选地包括用于病毒DNA整合的序列。出于两个主要原因，使3'LTR在U3区至少缺失启动子以提供SIN载体(自灭活)，而不扰乱基因转移所需的功能：第一，避免DNA整合到基因组后寄主基因的反式激活；第二，允许逆转录后病毒顺式序列的自灭活。可选地，来自驱动基因组转录的5'-LTR的tat依赖性U3序列被非内源启动子序列置换。因此，在靶细胞中，将只从内部启动子转录序列(转基因)。
2.病毒RNA衣壳化所必需的ψ区。
3.RRE序列(REV反应元件)，其允许在结合Rev蛋白后将病毒信使RNA从细胞核输出到胞质溶胶中。
4.促进入核转运的DNA flap元件(cPPT/CTS)。
5.可选地，转录后调控元件，特别是改善树突细胞中抗原多肽表达的元件，例如WPRE顺式激活序列(旱獭乙型肝炎病毒应答后元件)也被添加以优化mRNA的稳定性(Zufferey等，1999年)；基质或支架附着区(SAR和MAR序列)，例如免疫球蛋白κ基因的基质或支架附着区(Park F.等，Mol Ther，2001年；4：164-173)。
本发明的慢病毒载体是非复制性的(复制缺陷)，即载体和慢病毒载体基因组被认为适合于减轻对复制型感染性慢病毒的顾虑，特别是在施用后不能从感染的寄主细胞形成新的颗粒出芽。这可以公知的方式实现，作为慢病毒基因组中缺失gag、pol或env基因，或者它们作为“功能性基因”缺失的结果。因此，gag和pol基因仅以反式提供。这也可以通过缺失颗粒形成所需的其他病毒编码序列和/或顺式作用遗传元件来实现。
“功能性”指基因被正确转录和/或正确表达。因此，如果在该实施方式中存在的本发明慢病毒载体基因组中含有gag、pol或env的序列，则它们各自不被转录或不完全转录；表述“不完全转录”指转录物gag、gag-pro或gag-pro-pol的改变，这些中的一个或几个未被转录。参与慢病毒复制的其他序列也可以在载体基因组中突变，以达到此种状态。逆转录病毒载体的复制缺失应与慢病毒基因组的复制区分开来。实际上，如前所述，慢病毒基因组可含有复制起点，确保慢病毒载体基因组的复制，而不必确保载体颗粒的复制。
为获得根据本发明的慢病毒载体，载体基因组(作为载体质粒)必须在颗粒或假颗粒中衣壳化。因此，除了包膜蛋白外，慢病毒蛋白必须与载体基因组一起反式提供到生产系统中(特别是产生细胞中)的载体基因组中，可以使用至少一个衣壳化质粒，该衣壳化质粒携带gag基因和pol慢病毒基因或整合缺陷型pol基因，优选缺少Vif-、Vpr、Vpu-和Nef-辅助基因的编码序列的一些或全部，并且可选地缺乏Tat(对于HIV-1慢病毒载体)。
使用其他质粒，该质粒携带编码包膜假型化蛋白的多核苷酸，该包膜假型化蛋白被选择用于假型化慢病毒载体颗粒。
在一个优选的实施方式中，包装质粒仅编码病毒颗粒合成所必需的慢病毒蛋白。因此移除了附属基因(accessory gene)，其中质粒中附属基因的存在可引起安全性问题。因此，被反式引入包装的病毒蛋白分别如源自HIV-1的那些病毒蛋白所示：
1.用于构建基质(MA，具有表观分子量p17)的GAG蛋白、衣壳(CA，p24)和核衣壳(NC，p6)。
2.POL编码的酶：整合酶、蛋白酶和逆转录酶。
3.当TAT是启动LTR介导的转录所必需时，TAT和REV调节蛋白；如果5'LTR的U3区被驱动tat非依赖性转录的启动子所置换时，则TAT表达可被省略。REV可以被修饰并因此用于例如重组蛋白(该重组蛋白能够识别载体基因组中置换RRE序列的结构域)中，或者用作能够通过其RBD(RNA结合结构域)结合RRE序列的片段。
为避免由包装质粒包含的基因产生的mRNA在病毒颗粒中的任何包装，从包装质粒中除去ψ区。将异源启动子插入质粒中以避免重组问题，并将聚腺苷酸(poly-A)尾部加入到编码蛋白质的序列的3'端。上文已公开了合适的启动子。
如本文所公开的，在内部启动子的控制下，包膜质粒编码本文公开的用于假型化的包膜蛋白。
用于制备本发明的慢病毒载体颗粒的任何或所有所述质粒，可在编码蛋白质的区段中进行密码子优化(CO)。在哺乳动物细胞、鼠或特别是人细胞中，优选进行根据本发明的密码子优化以改善质粒中包含的编码序列的翻译。根据本发明，密码子优化特别适用于直接或间接地改善载体颗粒的制备，或改善施用载体颗粒的寄主细胞对载体颗粒的摄入，或提高多核苷酸编码转移的效率，寄主转导细胞基因组中的所述多核苷酸编码疟疾寄生虫的抗原多肽(转基因)。优化密码子的方法是本领域公知的，特别地，为此密码子优化使用可获得的程序来实现。说明了实施例所用编码序列的密码子优化。
在本发明的一个具体实施方式中，假型慢病毒载体也是或者可选地是整合型的，因此它能够将载体基因组及它所含多核苷酸整合到转导细胞或寄主(其已被施用该慢病毒载体)细胞的基因组中。
在本发明的另一个具体实施方式中，假型慢病毒载体也是或可选地是整合缺陷型的。在此种情况下，载体基因组及因此它所含重组多核苷酸不整合到转导细胞或寄主(其已被施用该慢病毒载体)细胞的基因组中。
本发明涉及免疫原性组合物中慢病毒载体的用途，其中表达的整合酶蛋白是缺陷型的，并且该慢病毒载体还包含多核苷酸，特别地，该多核苷酸编码至少一种携带疟原虫寄生虫的红前期抗原表位的抗原多肽。
“整合缺陷型”指整合酶(优选慢病毒来源)缺乏将慢病毒基因组整合到寄主细胞基因组中的能力，即整合蛋白质经突变而特异性地改变了其整合酶活性。
通过修饰编码整合酶的pol基因获得整合缺陷型慢病毒载体，产生编码整合缺陷型整合酶的突变pol基因，所述修饰的pol基因包含在衣壳化质粒中。专利申请WO2006/010834中描述了此种整合缺陷型慢病毒载体。因此，蛋白质的整合酶能力被改变；而衣壳化质粒的GAG、PRO和POL蛋白的正确表达和/或衣壳的形成以及因此载体颗粒的形成，以及在整合步骤之前或之后的病毒周期的其他步骤(例如逆转录、入核转运)保持完整。与含有相应野生型整合酶的慢病毒载体相比，当整合酶应启动的整合以整合步骤发生低于1/1000(优选低于1/10000)的方式被改变时，整合酶是所述缺陷型。
在本发明的一个具体实施方式中，缺陷型整合酶由第1类突变产生，优选满足缺陷型整合酶表达要求的氨基酸置换(单氨基酸置换)或短缺失。突变在pol基因内进行。这些载体可以在酶的催化结构域中携带具有突变D64V的缺陷型整合酶，其特异性地阻断整合步骤的DNA切割和连接反应。D64V突变使假型化HIV-1的整合降低至野生型的1/10,000，但保持其转导非分裂细胞的能力，从而允许有效的转基因表达。
pol基因中适合影响HIV-1整合酶的整合酶能力的其他突变如下：H12N、H12C、H16C、H16V、S81R、D41A、K42A、H51A、Q53C、D55V、D64E、D64V、E69A、K71A、E85A、E87A、D116N、D116I、D116A、N120G、N120I、N120E、E152G、E152A、D-35-E、K156E、K156A、E157A、K159E、K159A、K160A、R166A、D167A、E170A、H171A、K173A、K186Q、K186T、K188T、E198A、R199C、R199T、R199A、D202A、K211A、Q214L、Q216L、Q221L、W235F、W235E、K236S、K236A、K246A、G247W、D253A、R262A、R263A和K264H。
在一个具体实施方式中，pol基因中的突变在以下位置D64、D116或E152中的任一处进行，或者在蛋白质的催化位点中的这些位置中的几个位置进行。在这些位置的任何置换都是合适的，包括上述那些。
提出的另一个置换是用氨基酸残基AAH置换氨基酸残基RRK(第262至264位)。
在本发明的一个具体实施方式中，当慢病毒载体是整合缺陷型时，慢病毒基因组还包含复制起点(ori)，其序列取决于要表达慢病毒基因组的细胞的性质。所述复制起点可以是真核生物来源，优选哺乳动物来源，最优选人来源。它也可以是病毒来源，特别是DNA环状附加体病毒来源，如SV40或RPS。本发明的一个有利的实施方式在本发明的慢病毒载体的慢病毒基因组中插入复制起点。实际上，当慢病毒基因组不整合到细胞寄主基因组中时(由于缺陷型整合酶)，慢病毒基因组在经历频繁细胞分裂的细胞中丢失；在免疫细胞(例如B或T细胞)中尤其如此。复制起点的存在确保每个细胞中存在至少一个慢病毒基因组(甚至在细胞分裂后)，从而使免疫应答效率最大化。
特别地，本发明的所述慢病毒载体的慢病毒载体基因组可衍生自HIV-1质粒pTRIPAU3.CMV-GFP或它的变体，该质粒于1999年10月11日保藏于CNCM(法国巴黎)，编号为I-2330(也在WO01/27300中描述)。此类变体的序列如SEQ ID NO：1或2所示。
当载体基因组衍生自这些特定质粒时，除了GFP编码片段之外或置换GFP编码片段，重组多核苷酸(其编码编码本申请所公开的疟原虫寄生虫的抗原多肽)的序列被插入该载体基因组中。GFP编码序列也可以被不同的标记(marker)置换。CMV启动子也可以被另一个启动子置换，特别是上文公开的启动子之一，特别是与转基因表达有关的启动子。
可选地，也可以缺失特定保藏的pTRIP载体中包含的WPRE序列。
通过所述质粒或通过其他方法转染合适的细胞(例如哺乳动物细胞或人细胞，例如293T细胞所示的人胚肾细胞)后，可以产生载体颗粒。在用于表达慢病毒颗粒的细胞中，所有或一些质粒可用于稳定地表达其编码多核苷酸，或者用于瞬时或半稳定地表达其编码多核苷酸。
产生的颗粒浓度可以通过测量细胞上清液的P24(HIV-1的衣壳蛋白)含量来测定。
一旦施用到寄主中，本发明的慢病毒载体感染寄主细胞，可能是特定细胞，这取决于对它进行假型化的包膜蛋白。感染导致慢病毒载体基因组释放到寄主细胞的细胞质中，并在细胞质中发生逆转录。一旦处于三链体形式(通过DNA flap)，慢病毒载体基因组被转运入细胞核，细胞核中编码疟疾寄生虫抗原多肽的多核苷酸通过细胞机器进行表达。当转导非分裂细胞(如DC)时，表达可以是稳定的。当转导分裂细胞(如B细胞)时，在慢病毒基因组中缺乏复制起点的情况下，由于核酸稀释和细胞分裂，表达是暂时的。通过提供复制起点(确保在细胞分裂后慢病毒载体基因组适当地扩散到子细胞中)，表达可以延长。通过在载体基因组中插入MAR(基质结合区，Matrix Associated Region)或SAR(支架结合区，Scaffold Associated Region)元件，也可以增加稳定性和/或表达。
实际上，这些SAR或MAR区是富含AT的序列并且能够将慢病毒基因组锚定到细胞染色体的基质上，从而调节编码至少一种抗原多肽的多核苷酸的转录，特别是刺激转基因的基因表达和改善染色质可及性(chromatin accessibility)。
如果慢病毒基因组是非整合型，它不会整合到寄主细胞基因组中。然而，由转基因编码的至少一种多肽被充分表达并且足够长以进行加工，与MHC分子结合并最终导向细胞表面。根据编码疟疾寄生虫的抗原多肽的多核苷酸的性质，与MHC分子结合的至少一个多肽表位引发体液或细胞免疫应答。
除非另有说明，或除非技术上不相关，否则本申请中公开的关于慢病毒颗粒的结构或用途(特别是关于它们的包膜蛋白，或重组多核苷酸)的任何各种特征、实施方式或实例的特征可以根据任何可能的组合进行组合。
本发明还涉及用于给哺乳动物寄主单独施用的化合物的组合物，该化合物的组合物至少包括：
(i)本发明的慢病毒载体颗粒，该颗粒用第一种确定的异源病毒包膜假型化蛋白进行假型化；此种第一种假型化蛋白可来自VSV的新泽西株；
(ii)与(i)中的慢病毒载体颗粒分开提供的本发明的慢病毒载体颗粒，该颗粒用第二种确定的异源病毒包膜假型化蛋白进行假型化，所述第二种确定的异源病毒包膜假型化蛋白不同于第一种异源病毒包膜假型化蛋白；此种第二种假型化蛋白可来自VSV的印第安纳株。
本发明还涉及一种多核苷酸，该多核苷酸是小鼠密码子优化的核酸，所述核酸编码疟原虫寄生虫的红前期抗原，其中，所述多核苷酸选自：
-a/伯氏疟原虫CSP的SEQ ID NO：10、伯氏疟原虫TRAP的SEQ ID NO：19、伯氏疟原虫ICP的SEQ ID NO：28、伯氏疟原虫的Falcilysin的SEQ ID NO：37、伯氏疟原虫GPI锚定蛋白P113的SEQ ID NO：57、伯氏疟原虫成孔样蛋白SPECT2，用于伯氏疟原虫的的SEQ ID NO：48、伯氏疟原虫蛋白Ag40 11-09的SEQ ID NO：66和伯氏疟原虫蛋白Ag45 11-10的SEQ IDNO：75，或者，
-b/恶性疟原虫CSP的SEQ ID NO：13、恶性疟原虫TRAP的SEQ ID NO：22、恶性疟原虫ICP的SEQ ID NO：31、恶性疟原虫Falcilysin的SEQ ID NO：40、恶性疟原虫成孔样蛋白SPECT2的SEQ ID NO：51、恶性疟原虫GPI锚定蛋白P113的SEQ ID NO：60、恶性疟原虫蛋白11-09的SEQ ID NO：69和恶性疟原虫蛋白11-10的SEQ ID NO：78，或者，
-c/间日疟原虫CSP的SEQ ID NO：16、间日疟原虫TRAP的SEQ ID NO：25、间日疟原虫ICP的SEQ ID NO：34、间日疟原虫Falcilysin的SEQ ID NO：43、间日疟原虫成孔样蛋白SPECT2的SEQ ID NO：54、间日疟原虫GPI锚定蛋白P113的SEQ ID NO：63、间日疟原虫蛋白11-09的SEQ ID NO：72和间日疟原虫蛋白11-10的SEQ ID NO：81。
当涉及编码伯氏疟原虫抗原的多核苷酸时，已经进行了上述序列中反映的密码子优化以在小鼠中表达。当涉及编码恶性疟原虫或间日疟原虫抗原的多核苷酸时，已进行了上述序列中反映的密码子优化在人细胞中表达。
本发明还涉及上述多核苷酸的用途，用于设计核酸的替代形式也适用于制备本发明的载体，其中，由此获得的核酸是经修饰的DNA的RNA，例如苏糖核酸(TNA)、乙二醇核酸(GNA)、肽核酸(PNA)、具有已知构型的锁核酸(LNA)或乙烯核酸(ENA)或环己烯基核酸(CeNA)或它们的杂合体或组合。特别是当进行本发明的载体基因组的制备时，可以使用杂合分子，其中，编码本文公开的疟疾寄生虫的抗原多肽的多核苷酸由上述公开的序列形式之一表达。根据本发明的一个实施方式，载体基因组的核苷酸序列是嵌合序列，它包含用于转录抗原多肽的经修饰的核酸。
在本发明的另一个实施方式中，可能与上述公开的核酸替代形式组合，编码抗原多肽的多核苷酸在结构上被修饰和/或化学修饰。说明一下，多核苷酸在其5'区域中包含Kozak共有序列。本文特别说明了包含Kozak共有序列的此类多核苷酸。非慢病毒来源的其他核酸序列可在载体基因组中存在IRES序列(内部核糖体进入位点)，其适用于启动多肽合成WPRE序列作为转录后调控元件以稳定产生的RNA。
根据本发明的另一个实施方式，如果多个异源多肽由一个载体基因组编码，则可选地编码序列可通过接头部分分开，该接头部分是基于核酸的分子或基于非核酸的分子。此种分子可以是功能化的接头分子，旨在识别与其连接的3'官能化核酸。或者，适合用作接头的序列可以是编码自体裂开(self-cleaving)肽的核酸，例如2A肽。
本发明涉及如此描述的多核苷酸的集合，其中，每个多核苷酸编码如本文所述的用于本发明目的的一种以上疟疾寄生虫的抗原多肽，前提是这些多核苷酸的集合适用于制备本发明的化合物的组合物的活性成分。
本发明还涉及如此公开的多核苷酸的用途，用于制备慢病毒载体(特别是基于HIV-1的载体)的集合，其中，载体在其基因组中包含这些多核苷酸中的至少一种，前提是该慢病毒载体的集合使得能够表达由a/、b/和c/中选择的一个亚组的多核苷酸编码的所有抗原多肽。
本发明的其他特征和性质，包括在上述实施方式中使用的特征，将在随后的实施例和附图中描述，并因此可用于表征本发明。
附图说明
图1：用于生产慢病毒颗粒的质粒的图示。
图2：用5×107TU的VSVIND假型化慢病毒颗粒肌内免疫C57BL/6小鼠(n＝5)，该慢病毒颗粒编码抗原、CSP、Celtos SPECT、HSP20和Ag13。作为保护性的阳性对照，通过静脉内注射用50k经辐照的子孢子免疫小鼠。免疫后30天，用10,000个生物发光子孢子(被皮下微量注射到小鼠足垫中)攻击动物。两天后通过生物发光定量肝脏中的寄生虫负荷，如CSP、Celtos和Ag13的图片所示。该图显示了肝脏感染的量化，肝脏感染表示为log(平均辐射度)(方块)。虚线表示未感染区域的背景信号(Bk)的平均值。*P<0.05和***P<0.001(ANOVA)。
图3：C57BL/6小鼠(每组n＝5)用5×107TU携带Ag13(阴性对照)和CSP(阳性对照)的VSVIND LP免疫或者不进行免疫(幼稚型)。接受浓缩LP的组用50uL载体(Ag13im c和CSPim c)肌内接种在大腿肌肉中。接受非浓缩LP的组用700uL载体(Ag13 ip nc和CSP ip nc)腹膜内接种。免疫后30天，用5,000个表达荧光素酶的子孢子(皮下微量注射在小鼠足垫中)攻击动物。两天后通过生物发光定量肝脏中的寄生虫负荷，如图2所示。该图显示SPZ接种后两天肝脏中log(平均辐射度)的平均值和sd。虚线表示背景信号(Bk)的平均值。ns：不显著(ANOVA)。
图4：在CMV或B2M启动子(分别为CMVCSP和B2M CSP)的控制下，用1×107TU的非浓缩VSVIND CSP LP对C57BL/6小鼠(每组n＝4-5)进行腹膜内免疫或不进行免疫(幼稚型)。免疫后30天，用5,000个表达荧光素酶的子孢子(皮下微量注射在小鼠足垫中)攻击动物。两天后通过生物发光定量肝脏中的寄生虫负荷，如图2所示。该图显示SPZ接种后两天肝脏中log(平均辐射度)的平均值和sd。虚线表示背景信号(Bk)的平均值。*P<0.05；ns：不显著(ANOVA)。
图5：使4周龄和7周龄的C57BL/6小鼠(每组n＝4)适应3周(旧组)和3天(新组)。然后用1×107TU的非浓缩VSVIND B2M CSP或GFP LP对这些年龄匹配的组进行腹膜内免疫。免疫后30天，用5,000个表达荧光素酶的子孢子(皮下微量注射在小鼠足垫中)攻击动物。两天后通过生物发光定量肝脏中的寄生虫负荷，如图2所示。该图显示SPZ接种后两天肝脏中log(平均辐射度)的平均值和sd。虚线表示背景信号(Bk)的平均值。<*>P<0.05；ns：不显著(ANOVA)。
图6：使4周龄C57BL/6小鼠(每组n＝4)适应3周(旧组)并用不同剂量的非浓缩VSVIND B2M CSP(黑)或GFP(白)LP进行腹膜内免疫。免疫后30天，用5,000个表达荧光素酶的子孢子(皮下微量注射在小鼠足垫中)攻击动物。两天后通过生物发光定量肝脏中的寄生虫负荷，如图2所示。该图显示SPZ接种后两天肝脏中log(平均辐射度)的平均值和sd。虚线表示背景信号(Bk)的平均值。**P<0.05；<***>P<0.001；ns：不显著(ANOVA)。
图7：存储在Plasmodb中的伯氏疟原虫(Pb)和恶性疟原虫(Pf)的子孢子、红外期和红内期cDNA文库的分析。将每个表达序列标签(EST，expression sequence tag)的百分比相对于EST的总数归一化并显示累积。每个符号代表一个基因，按EST丰度(从高到低)排序，并表示为总EST的百分比。值得注意的是，约10％的基因(丰富最高)占总EST的约50％(虚线)。
图8：抗原的表达和表面定位。表达GFP的Pb子孢子用2％的PFA固定，并用0.1％的Triton X100透化(透化的)或者不用(活的)。寄生虫与所示免疫血清(1/50)在冰上孵育1小时，洗涤，并用Alexa Fluor 647标记的山羊抗小鼠二抗显色。然后通过细胞计数分析子孢子，如右图所示(表面，使用活的非透化SPZ染色；透化的，使用固定和透化的SPZ染色)或通过荧光显微镜，如图中所示。注意，CSP和抗原9-6通过细胞计数和显微镜两者呈现表面图案染色。
图9：靶向筛选保护性抗原。使4周龄的C57BL/6小鼠(每组n＝5)适应3周，并用单剂量的1×107TU的非浓缩VSVIND B2M LP进行腹膜内免疫。免疫后30天，用5,000个表达GFP的子孢子(皮下微量注射在小鼠足垫中)攻击动物。通过流式细胞术检测寄生虫感染。该图显示了用所示疟原虫抗原免疫的log(寄生虫血症)的平均值(标线，圆圈代表个体小鼠)。粗体虚线表示GFP log正态分布的95％容许区间。具有低于容许区间下限的寄生虫血症的小鼠被认为是受保护的。顶部虚线是对照的平均值，底部虚线表示未感染的(NI)小鼠。
图10：一个或两个免疫剂量引起的保护作用的比较。使4周龄的C57BL/6小鼠(每组n＝5)适应3周，并用第一剂5×105TU的非浓缩VSVNJ B2M LP进行腹膜内免疫。第一次免疫后30天，动物接受第二剂1×107TU的非浓缩VSVIND B2M LP。30天后，用5,000个表达荧光素酶的子孢子(皮下微量注射在小鼠足垫中)攻击动物。通过流式细胞术检测寄生虫感染。该图显示了受攻击小鼠个体接种后第5天的log(寄生虫血症)，其中该小鼠接种两个免疫剂量(Squares，PB)。圆圈代表仅接受一次LP免疫剂量的小鼠(实验数据如图9所示)。标线代表Log(寄生虫血症)的平均值。粗体虚线表示GFP log正态分布的95％容许区间。低于容许区间下限的小鼠被认为是受保护的。Nl：未感染的小鼠。
图11：靶向筛选保护性抗原。使4周龄的C57BL/6小鼠(每组n＝5-10)适应3周并用第一剂5×105TU的非浓缩VSVNJ B2M LP进行腹膜内免疫。第一次免疫后30天，动物接受第二剂1×107TU的非浓缩VSVIND B2M LP。第三天后，用5,000个表达GFP的子孢子(皮下微量注射在小鼠足垫中)攻击动物。通过流式细胞术检测寄生虫血液感染。(a)上图显示了感染后第5天个体小鼠的log(寄生虫血症)。标线代表log(寄生虫血症)的平均值值。GFP组(保护对照)的平均值由虚线中间线表示。上部和下部虚线显示了GFP对照组的95％容许区间(灰色框)。CSP组是保护的阳性对照。NI(未感染＝感染后第10天没有寄生虫血症，基于我们的寄生虫血症定量方法的检测限)。黑色圆圈代表抗原，其中，平均log(寄生虫血症)显著减少，因此被认为是保护性的(ANOVA)。(b)下图表示受保护小鼠的百分比(低于95％容许区间的动物的百分比)。黑条表示保护性抗原(Fisher精确检验)。*P<0.05，**P<0.01，****P<0.0001。
图12：伯氏疟原虫保护性抗原的结构。根据右侧的条码，保守结构域和功能性结构域用框表示。GPI(糖基磷脂酰肌醇)、TSR(血小板反应蛋白I型重复)、MACPF(膜攻击复合物/穿孔素)。
图13：保护性抗原在疟原虫物种中是保守的。通过按log期望的多序列比对(MUSCLE，MUltiple Sequence Comparison by Log-Expectation)对来自感染啮齿类动物的伯氏疟原虫、感染猕猴的食蟹猴疟原虫和感染人的恶性疟原虫和间日疟原虫寄生虫的保护性直系同源抗原的氨基酸序列进行比对。垂直黑条表示四种疟原虫物种中保守的相同氨基酸，短的深灰色条代表重复区域，以及短浅灰色条代表用于比对的插入空位。
图14：下选(down-selected)保护性抗原与次优(sub-optimal)剂量CSP的组合诱导的保护作用。将小鼠免疫两次，间隔4周，用次优剂量的CSP(第一次免疫中的5×105TU非浓缩VSVNJ B2M LP和第二次免疫中的5×106TU非浓缩VSVIND B2M LP，白色三角形，CSP)和通常剂量的保护性疟原虫抗原(CSP+11-03、+11-05、+11-06、+11-07、+11-09和+11-10；三角形)。作为阴性对照小鼠，用通常的两剂GFP免疫。第二次免疫剂量后4周，用5,000个子孢子攻击动物。
图15：多基因组合诱导的无菌保护。以7×个体剂量(individual dose)(1剂量＝第一次免疫中的5×105TU非浓缩VSVNJ B2M LP/第二次免疫中的1×107TU非浓缩VSVIND B2MLP)的对照抗原AL11-荧光素酶(Luc，白色三角形)、个体剂量的CSP加6×Luc(灰色三角形)或个体剂量的CSP和6种保守PE抗原(11-05、11-06、11-07、11-09、11-10和18-10；黑色三角形，7cPEAg)将小鼠免疫两次，间隔4周。在第二次免疫剂量后4周，用5,000个GFP SPZ攻击小鼠。两个图均显示了攻击后第5天个体的log(寄生虫血症)。(a)该图显示了三次独立实验的合并结果。受无菌保护小鼠数量/受攻击小鼠数量：7×Luc(0/21)、1×CSP 6×Luc(1/20)和1×7cPEAg(18/21)。(b)在子孢子攻击前3天和1天，给1×7cPEAg免疫小鼠注射400μg对照(Ctr)、CD4耗竭(a-CD4+，克隆GK1.5)和CD8耗竭(a-CD8+，克隆2.43)单克隆抗体。GFP数据来自图11中的实验(灰色圆圈)。受无菌保护小鼠数量/受攻击小鼠数量：7×Luc(0/7)、1×CSP6×Luc(1/7)和1×7cPEAg(ctr，7/7；a-CD8，0/7和a-CD4，7/7)。请注意，CD8+细胞的耗竭消除了无菌保护。*P<0.05，**P<0.01，****P<0.0001(ANOVA)。
图16：单次免疫剂量中多基因组合诱导的无菌保护。(a)以7×个体剂量(1剂量＝第一次免疫中的5×105TU非浓缩VSVNJ B2M LP/第二次免疫中的1×107TU非浓缩VSVIND B2MLP)的对照抗原AL11-荧光素酶(Luc，白色三角形)、个体剂量的CSP加6×Luc(灰色三角形)或个体剂量的CSP和6种保守PE抗原(11-05、11-06、11-07、11-09、11-10和18-10；黑色三角形，2im 7cPEAg)将小鼠免疫两次，间隔4周。或者，仅给小鼠施用第二次个体免疫剂量(1×107TU)的CSP和6种保守的PE抗原(11-05、11-06、11-07、11-09、11-10和18-10；灰色菱形，1im 7cPEAg)。在第二次免疫剂量后4周，用5,000个GFP SPZ攻击小鼠。图显示了攻击后第5天个体的log(寄生虫血症)。黑条是log(寄生虫血症)的平均值。受无菌保护小鼠数量/受攻击小鼠数量：Luc(0/7)、CSP(0/7)、2im 7cPEAg(6/7)和1im 7cPEAg(6/7)。(b)以9×个体剂量(1剂量＝第一次免疫中的1×107TU非浓缩VSVIND B2M LP)的对照抗原AL11-荧光素酶(Luc，黑色菱形)或个体剂量的CSP+7种保守PE抗原(11-05、11-06、11-07、11-09、11-10、18-10、30-03A和30-03B；灰色菱形，8cPEAg)将小鼠免疫一次。在子孢子攻击前3天和1天，给8cPEAg免疫小鼠注射400μg对照(Ctr)、CD4耗竭(a-CD4+，克隆GK1.5)和CD8耗竭(a-CD8+，克隆2.43)单克隆抗体。单次免疫剂量后4周，用5,000个GFP SPZ攻击小鼠。该图显示了攻击后第5天个体的log(寄生虫血症)。黑条是log(寄生虫血症)的平均值。受无菌保护小鼠数量/受攻击小鼠数量：7×Luc(0/7)和8cPEAg(ctr，6/7；a-CD8，0/7和a-CD4，7/7)。请注意，CD8+细胞的耗竭消除了无菌保护。*P<0.05；**P<0.01；****P<0.0001；ns：P>0.05(ANOVA)。
图17：5种PE抗原的最小组合诱导的无菌保护。(a)以个体剂量×圆圈中所示数量(1剂量＝第一次免疫中的5×105TU非浓缩VSVNJ B2M LP/第二次免疫中的1×107TU非浓缩VSVIND B2M LP)将小鼠免疫两次，间隔4周。例如，对于对照抗原AL11-荧光素酶(LUC)，用5×个体剂量免疫动物。所有组均接受5剂，除了接受7剂LP(7PEAg)的阳性对照保护。在第二次免疫剂量后4周，用5,000个GFP SPZ攻击小鼠。条代表受无菌保护小鼠的百分比。受无菌保护小鼠数量/受攻击小鼠数量显示在条的右侧。**P<0.01(Fisher精确检验)。(b)以个体剂量(1剂量＝第一次免疫中的5×105TU非浓缩VSVNJ B2M LP/第二次免疫中的1×107TU非浓缩VSVIND B2M LP)的对照抗原GFP(GFP，黑色圆圈)或者个体剂量的CSP、TRAP、18-10、11-09或11-10(灰色三角形)将小鼠免疫两次，间隔4周。在子孢子攻击前3天和1天，给免疫小鼠注射400μg对照(Ctr)、CD4耗竭(a-CD4+，克隆GK1.5)和CD8耗竭(a-CD8+，克隆2.43)单克隆抗体。第二次免疫剂量后4周，用5,000GFP SPZ攻击小鼠。图显示了攻击后第5天的log(寄生虫血症)的平均值±sd。*P<0.05；ns：P>0.05(ANOVA)。(c)根据预测CD8 T细胞表位的存在和遵循抗原示意图所示的保守结构域，分离5种下选的保护性抗原。示意性蛋白质上方的图表示使用SYFPEITHI(得分)和IEDB ANN IC 50(nM)预测的结合H2Kb(8aa)和H2Kd(9aa)MHC I类分子的表位分布。示意性蛋白质左侧的图表代表由这些构建体诱导的保护，其中，条是攻击后第5天的log(寄生虫血症)的平均值±sd。显示抗原11-09的数据来自图11。虚线表示图11中计算的对照的容许区间的下限。*P<0.05；ns：P>0.05(ANOVA)。(d)预测与CD8+T细胞依赖性PE抗原区段中的MHC I类分子结合的最佳表位的相关性和从17c获得的平均保护活性。圆圈显示了使用IEDBANN软件的IC50和使用SYFPEITHI对得分值进行平方。虚线表示Luc对照的平均值。
图18：保守氨基酸区域中CD8 T细胞表位的聚类(clustering)和预测的Pf表位与HLAA02：01的结合。通过按log期望的多序列比对(MUSCLE)对来自感染啮齿类动物的伯氏疟原虫、感染猕猴的食蟹猴疟原虫和感染人的恶性疟原虫和间日疟原虫寄生虫的保护性直系同源抗原的氨基酸序列进行比对。垂直黑条表示四种疟原虫物种中保守的相同氨基酸。该图显示了预测结合H2Kb(8aa)和H2Kd(9aa)MHC I类分子的Pb表位或预测结合HLAA02：01(9聚体)的Pf表位的分布，其使用SYFPEITHI(得分)和IEDB ANN IC50(nM)。在肽和p2-微球蛋白(Score)存在下，在MHC I类分子的稳定化的测定中检测最佳预测的HLA结合物。得分100对应于阳性对照肽的结合。注意保守氨基酸区域中表位的聚类。
图19：用于产生VSV假型慢病毒颗粒的质粒：pTRIP CMV GFP
质粒的序列由以下功能区构成，其中，顺式激活慢病毒区源自HIV基因组，驱动蛋白质表达的启动子(GFP)是CMV：
质粒中提供载体基因组的插入物组成如下：LTR-Ψ-RRE-cPPT/CTS-CMV-GFP-WPRE-AU3LTR，其中：
LTR是长末端重复
Psi(ψ)是包装信号
RRE是Rev反应元件
CMV是早期CytoMegaloVirus启动子
cPPT是中央多嘌呤管道，其中，从cPPT至CTS的核苷酸区段形成flap序列
CTS是中央终止序列
WPRE是旱獭肝炎病毒后调控元件(Post Regulatory Element)
核苷酸序列如SEQ ID NO：1所示。
图20：用于产生VSV假型慢病毒颗粒的替代质粒(图19质粒的替代质粒)：pTRIPB2M GFP
提供载体基因组的质粒中的插入物的组成如下：LTR-Ψ-RRE-cPPT/CTS-B2M-GFP-WPRE-AU3LTR。
核苷酸序列如SEQ ID NO：2所示。
图21：用于产生VSV假型慢病毒颗粒的质粒：包装8.74质粒
该质粒在CMV启动子的控制下提供HIV-1慢病毒所需的GAG和POL编码序列。
核苷酸序列如SEQ ID NO：3所示。
图22：用于产生VSV假型慢病毒颗粒的质粒：衣壳化质粒pCMV-VSV-INDco
包膜蛋白是印第安纳株的VSV-G，编码序列已经过小鼠密码子优化。
核苷酸序列如SEQ ID NO：4所示。
图23：用于产生VSV假型慢病毒颗粒的替代质粒(图22质粒的替代质粒)：衣壳化质粒pCMV-VSV-NJco
包膜蛋白是新泽西株的VSV-G，并且编码序列已经过小鼠密码子优化。
核苷酸序列如SEQ ID NO：5所示。
具体实施方式
下表提供了序列表中所包含序列的列表和标识号。
与一些上表中公开序列有关的附加信息在下表中提供。
(1)：https：//www.ncbi.nlm.nih.gov/biosample/SAMN01737342
(2)：https：//www.ncbi.nlm.nih.gov/biosample/SAMEA2394724
(3)：https：//www.ncbi.nlm.nih.gov/biosample/SAMN00710434
实施例
为了解决鉴定保护性抗原的复杂问题，发明人设计了功能性筛选以使用啮齿动物疟疾模型鉴定和组合新型PE保护性抗原，在该模型中，小鼠(C57BL/6)对伯氏疟原虫(Pb)子孢子感染极其敏感。在该模型中，由活的经辐照的子孢子诱导的灭菌保护由抗体介导，并且主要由分别针对子孢子和红外期的CD8 T细胞应答介导。发明人的筛选策略基于四个主要特征设计：i)基于丰度、直系同源性、预测的拓扑和功能，参数化选择55种PE抗原；ii)密码子优化的抗原的合成以避免富含AT的疟原虫序列并最大化在哺乳动物细胞中的表达；iii)使用基于HIV的慢病毒载体进行免疫，该载体引发强烈的体液应答和细胞应答11,12；以及iv)在严重的子孢子攻击(皮下接种在免疫小鼠中)后测量保护作用。
1、设置筛选参数
在旨在验证中等通量下筛选抗原策略的可行性的概念验证(proof-of-concept)实验中，发明人订购了Pb CSP(一种已知的保护性抗原)的小鼠密码子优化的合成基因(SEQID NO：11)和其他4种其他子孢子抗原(Celtos，SPECT，HSP20和Ag13)，它们之前与保护相关13。将合成的疟原虫基因克隆到pTRIP载体质粒中，该质粒通过即刻(immediate)早期巨细胞病毒启动子(CMV)和旱獭肝炎病毒(WPRE)的转录后调控元件驱动它们在哺乳动物细胞中的表达(图1，SEQ ID NO：1)。这两种元件确保了抗体在体内多种小鼠细胞中的强烈表达。通过用三种辅助质粒(编码单独包装功能)瞬时共转染HEK293T细胞，制备HIV-1衍生的慢病毒颗粒，该三种辅助质粒为：含有合成疟原虫基因的pTRIP载体质粒、编码来自印第安纳(VSVIND)血清型或新泽西(VSVNJ)血清型水疱性口炎病毒的糖蛋白G的包膜表达质粒，以及p8.74衣壳化质粒(图1)。该共转染产生整合型但复制缺陷的假型慢病毒颗粒，其能够转导分裂和非分裂细胞(包括树突细胞)并诱导有效的细胞6和体液7记忆应答。在共转染后48小时收集颗粒，并通过定量PCR在HeLa细胞中滴定每批载体。该功能性滴定方法给出了能够转移1个基因拷贝/细胞的颗粒浓度，并以转导单位(TU)/ml表示。质粒序列显示在图中，它们的序列在序列表中提供。
用单次肌内剂量的5e7 TU的超速离心浓缩的VSVIND假型慢病毒颗粒(LP)免疫5只小鼠的组。免疫后30天，用10,000个生物发光子孢子(足垫内皮下接种)攻击小鼠。两天后，通过生物发光测量肝脏中的寄生虫负荷。值得注意的是，在用10,000个生物发光子孢子攻击后，CSP免疫使肝脏中寄生虫负荷减少了15倍，而用50,000个经辐照子孢子攻击的静脉注射免疫使动物的肝脏中寄生虫负荷减少了5倍，这是我们的保护作用的金标准(图2)。这一初步的且良好的结果证实了本方法从功能上鉴定新的保护性抗原的高性能，并显示了扩大我们的检测样品的可行性。
发明人接下来旨在将这些最佳实验条件转换为更大规模筛选的条件。此种转换包括：验证非浓缩LP的使用、驱动疟原虫抗原表达的最佳启动子的选择，以及免疫剂量。
检测的第一个参数是非浓缩LP(而非浓缩LP，以避免LP生产方案中昂贵且耗时的超速离心浓缩步骤)的使用，其需要大量非浓缩的LP悬浮液。图3显示，相同剂量(5×107TU)的肌内注射的浓缩LP(CS im c，50μL)诱导的保护和腹膜内注射的非浓缩LP(CS ip nc，700μL)诱导的保护之间没有显著差异。免疫后30天，用5,000个子孢子进行攻击，通过生物发光评估肝脏感染的减少，来测量保护作用。作为保护的阴性对照，发明人使用用Pb Ag13免疫的小鼠，其先前被确定为非保护性抗原(图1)。
接下来对两个启动子进行了检测，以确定哪一个使用密码子优化的Pb CSP诱导了最佳保护。发明人比较了强和组成型巨细胞病毒(CMV)启动子与人β-2微球蛋白(B2M)启动子的使用，该启动子指导许多细胞类型中(特别是在树突细胞中)的基因表达。图4显示，在1×107TU的非浓缩LP的免疫剂量下使用B2M启动子，CSP诱导的保护稍好(尽管非统计学显著)。因此，发明人在我们的构建体中进一步采用了该启动子。
如图4所示，在优化期间，发明人观察到使用相同LP原液(stock)的CSP诱导保护的一些变化。发明人探讨此种变化是否可归因于小鼠适应过程，包括小鼠微生物群的改变。为了检验该假设，在免疫接种之前从Elevage Janvier(4周龄)购买一组小鼠，在动物设施中饲养3周(旧组)。购买第二组小鼠(7周龄)并在免疫前3天放入笼中(新组)。两组均用1×107TU的非浓缩LP进行腹膜内免疫。如图5所示，与适应3天小鼠相比，适应3周的小鼠得到显著且更均一的保护。因此，发明人在我们随后所有实验中采用了此适应期。
接下来，检测最佳保护性免疫剂量，范围为1×108至1×105TU的B2M CSP非浓缩LP。如图6所示，使用107和108TU观察到显著的保护，并且使用1×107TU的免疫剂量观察到最佳保护活性。在该实验中，如GFP组中所证明的，发明人还观察到随着时间的推移SPZ感染性逐渐丧失，这是由于使用单个SPZ原液来攻击所有动物。为了减少由于多次操作原液储存管而引起的温度变化，注射操作之间将储存管保持在冰上，在随后的攻击中发明人为每组制备了SPZ原液，并且此种变化消失了。
总之，基于CSP建立免疫方案，该方案依赖于给7周龄的C57BL/6小鼠单次腹膜内注射107TU的非浓缩VSVIND B2M LP，在动物设施中适应3周。在汇总数据中，使用5,000个表达荧光素酶的SPZ进行皮下攻击，如通过生物发光成像所评估的，该方案使得平均寄生虫肝负荷降低约5倍。
然而，此种检测寄生虫的生物发光方法存在一些缺点，例如，麻醉和发光底物的使用和相关成本、分析能力有限(每次采集少数动物)、耗时且预测无菌保护的灵敏度不够。因此，发明人决定使用荧光寄生虫通过流式细胞术在接种后第4、5、6和10天测量寄生虫血症来检查保护作用。发明人分析了至少100,000个红细胞，其提供的检测寄生虫血症的灵敏度为0.001％。在第4天至第6天，寄生虫在血液中呈指数增长，因此可以使用线性回归拟合寄生虫血症的log变换，其中，斜率表示每天寄生虫复制的时间。因此，发明人使用该参数来确定免疫是否影响血液中寄生虫的生长。为了量化保护，发明人使用接种后第5天的log(寄生虫血症)。这代表了肝脏感染的间接测量，并且因为记录了更多的受感染血细胞事件，比第4天的测量更稳健。最后，发明人确定，如果在接种后10天未检测到感染的红细胞，则免疫小鼠是受无菌保护的。在定义免疫方案和定量寄生虫感染的方法后，发明人开始筛选下选抗原的保护活性。
2、抗原的参数化选择
通过使用PlasmoDB(www.plasmodb.org)合并蛋白质组学和转录组学数据，发明人确定了三种不同疟原虫物种的疟原虫红前期表达的约9000个基因(唾液腺子孢子和红外期)，其中有3654个恶性疟原虫(Pf)的同线性直系同源物(syntenic ortholog)，Pf是最致命的人类感染疟原虫物种。通过分析病原体转录物的所有组成成分，如通过在不同阶段和疟疾寄生虫种类的cDNA文库中获得的表达序列标签(EST)的量的推断，发明人已观察到，EST总量的约50％来自这些文库中代表的仅约10％基因，相当于这些文库中的约100个基因(图7)。因此，通过关注约100最丰富的转录基因，发明人可以靶向给定寄生虫阶段的约50％的推定(待翻译的)抗原质量。因此，发明人选择了约50个大量转录的基因，该基因编码以高概率在寄生虫/感染细胞表面表达/出现的保守蛋白，优先考虑通过IEDB MHC结合算法预测的含有T细胞表位的候选物(http://tools.iedb.org/mhci/)。将Kozak共有序列(一种翻译起始位点)添加到这些下选基因中，然后通过MWG Eurofins(在图中列出)进行哺乳动物密码子优化和合成。然后将这些合成的密码子优化的下选疟原虫基因克隆到B2MpTRIP质粒中，并产生作为非浓缩的VSVIND LP。
3、使用单剂量LP的保护性抗原的第一次筛选
通常，通过实验检测6至10种疟原虫抗原，具有阴性(GFP)和阳性(CSP)保护对照。免疫后三周后，在透化和非透化的子孢子上检测免疫血清，从而确定：(i)寄主体液应答的效率，从而确定慢病毒传递的抗原的免疫原性；以及(ii)寄生虫抗原的定位(表面vs细胞内)。如图8所示，发明人用推定的GPI锚定抗原免疫小鼠，通过流式细胞术和免疫荧光(CSP和9-6)鉴定表面抗原。GFP和CSP组的血清分别作为细胞内和表面抗原定位的阳性对照。
免疫后4周，用5,000个表达荧光素酶的子孢子(皮下微量注射在小鼠足垫中)攻击动物。通过流式细胞术确定寄生虫血症。为了定义保护，对所有GFP组(感染后第5天，n＝35)的寄生虫血症进行log转换、合并、计算95％容许区间(图9)。与GFP组的平均log(寄生虫血症)相比，低于容许区间的下限(其表示寄生虫血症减少约8倍)的所有动物被认为是受保护的。作为阳性对照，43％动物(15/35)通过CSP免疫受保护，与GFP组相比平均减少约5倍。9％(3/35)在子孢子攻击后成为受无菌保护的。
在检测的第一组43种抗原中(图9)，我们鉴定了9种PE抗原，它们保护至少五只免疫小鼠中的一只(黑色圆圈；07-03、09-06、10-05、10-10、12-03、12-04、12-05、12-07和13-08)。其中，三个也被鉴定为子孢子表面抗原(09-06、10-05和10-10)。
为了验证筛选的稳健性，发明人选择了4种保护性抗原(CSP、09-06、10-05和07-03)、6种非保护性抗原(GFP、09-07、07-05、07-06、06-06和10-06)，发明人施用一剂5×105TU的非浓缩VSVNJ B2M LP并且一个月后施用第二剂1×107TU的VSUIND B2M LP，而不是仅施用一次免疫剂量。如图10所示，当发明人比较一个(圆圈，图9的数据)和两个免疫剂量(正方形，PB)时，发明人观察到三种感染概况。对于非保护性抗原GFP(09-07和07-05)，第二剂LP不像预期的那样改变感染概况。对于保护性抗原CSP(***P<0.001)(09-06、10-05和07-03)，如预期的那样，第二剂LP增加受保护小鼠的数量和/或降低平均寄生虫血症。值得注意的是，如用一剂LP免疫评估的，非保护性抗原07-06、06-06和10-06当在小鼠中施用两次时显示出强烈的保护活性，包括无菌保护的小鼠(PB7-6)。
这些结果验证了我们用单次免疫剂量检测到的一些保护性抗原，但也表明在我们的第一次筛选中有一些良好的保护性抗原未被检测到，因此决定使用两次免疫剂量重复筛选。
4、使用两剂LP进行第二次保护性抗原筛查
使用两种免疫剂量方案，通过功能性筛选55种下选的疟原虫PE抗原的保护活性，发明人鉴定了16种抗原，该16种抗原在每组中至少保护了一只免疫小鼠。与用GFP免疫的动物相比，在分析受保护小鼠的数量(Fisher精确检验)或log(寄生虫血症)的平均值值(ANOVA)时，这16种抗原中的7种(图11中的黑色圆圈/黑条)显示出了显著的保护作用。
当单独与我们的保护标准CSP(图11)进行比较时，它们都表现出相似或较差的保护活性。其中五种(11-05、11-06、11-07、30-03和18-10)是具有指定功能的分子，两种(11-09和11-10)是没有预测功能的蛋白质。这些Pb保护性抗原的结构显示在图12中，并且这些蛋白质与分别来自人感染寄生虫(恶性疟原虫(Pf)和间日疟原虫(Pv))和猕猴感染寄生虫(食蟹猴疟原虫)的直系同源的比对显示在图13中。如表I所示，直系同源之间相同氨基酸的百分比为：75％至38％(Pb vs Pf)、78％至33％(Pb vs Pv)和79％至26％(Pf vs Pv)。最保守的基因(>50％同一性)是30-03、11-09、11-10和11-06直系同源。具有分歧的(divergent)重复序列的抗原在比对中因插入空位而被罚分，表现出较低同一性百分比。
表I：通过CLUSTAL 2.1创建的百分比同一性矩阵。Pb抗原的氨基酸序列针对Pf和Pv taxid(有机体)进行了pBlast，使用MUSCLE(http：//www.ebi.ac.uk/Tools/msa/muscle/)将最佳匹配序列用于直系同源蛋白比对。该表显示了物种之间相同氨基酸的百分比。原始数据显示在图中。
在保护的递减顺序中，鉴定的第一抗原是TRAP14(血小板反应蛋白相关匿名蛋白；11-5)(SEQ ID NO：20和21)，其验证了我们的筛选方法，因为已知TRAP免疫在啮齿动物15和人类16中诱导保护。TRAP是I型跨膜蛋白，其携带两个细胞外粘附结构域(von Willebrand因子A型结构域和血小板反应蛋白I型结构域)，接着是富含脯氨酸的重复区。TRAP存储在微血管分泌囊泡中，并且在寄生虫激活后，TRAP易位至子孢子表面，在子孢子表面它充当固体基质和子孢子细胞质马达(motor)之间的接头。值得注意的是，抗TRAP抗体不损害寄生虫的运动性和感染性17，CD8+T细胞似乎介导由TRAP免疫介导的保护作用10,15,16,18。
鉴定的第二种保护性抗原是半胱氨酸蛋白酶抑制剂(ICP，18-10)19(SEQ ID NO：29和30)。ICP似乎通过CSP20,21的加工以及寄生虫肝内发育22参与子孢子的运动和感染能力。虽然蛋白质不具有膜定位的结构特征，但有证据表明该蛋白质位于子孢子的表面19,20。发表了关于寄生虫激活后蛋白质分泌的相反结果21,22。类似地，在抗ICP免疫血清存在下，存在关于子孢子体外抑制宿主细胞侵袭的相互矛盾的结果20,23。
鉴定的第三种保护性抗原是金属肽酶(Falcilysin/Bergheilysin，30-03)24(Bergheilysin A为SEQ ID NO：38，Bergheilysin B为SEQ ID NO：47，以及完整Bergheilysin ORF为SEQ ID NO：39)。该蛋白酶似乎参与寄生虫红内期血红蛋白的分解代谢25。最近描述了寄生虫血液感染期间的H-2Kb限制性CD8 T细胞表位25，表明CD8 T细胞可介导肝脏感染期间抗原30-03引发的保护作用。
第四种保护性抗原是最初在红内期16中描述的GPI锚定蛋白(P113，11-07)(SEQ IDNO：58和59)，并且也在PE阶段表达。P113似乎对肝脏感染很重要，而对于血液感染可有可无，但其确切的功能尚不清楚17。
第五种抗原是成孔样蛋白SPECT2(11-60)28(SEQ ID NO：49和50)。成孔样蛋白具有膜攻击复合物/穿孔素(MACPF)结构域，并且参与子孢子细胞穿透活动，对真皮中子孢子的进展29和肝脏中的吞噬作用存活是重要的30。
鉴定的第六种抗原是发明人称为11-09或Ag40(SEQ ID NO：67和68)的假定蛋白质。该蛋白质具有4至5个注释的跨膜结构域。编码抗原11-09的基因的缺失导致对肝脏中Pb寄生虫发育造成损害。
第七种抗原也是发明人称为11-10或Ag45(SEQ ID NO：76和77)的假定蛋白质。该蛋白质没有注释结构域，但具有带负电荷氨基酸的中心区域。最近，约氏疟原虫(另一种感染啮齿动物的疟原虫物种)的11-10直系同源物也被鉴定为保护性抗原21。编码抗原11-10的基因的缺失阻断了Pb子孢子侵入唾液腺并完全消除了子孢子感染肝脏的能力。
为确定基于CSP的保护是否可以通过抗原的组合进行加成或协同改善，发明人评估了在不存在或存在通常剂量的保护性抗原(5×105TU的VSVNJ/1×107TU的VSVIND B2M LP)的情况下，由次优剂量的CSP(5×105TU的VSVNJ/5×106TU的VSVIND B2M LP)引起的保护。将由CSP+保护性抗原诱导的此种保护作用与仅由这些抗原引发的保护作用(图11的数据)进行比较。作为阴性对照，发明人使用用常用剂量GFPLP免疫的动物。图14显示，与次优剂量的CSP(CSP+11-03、+11-10、+11-07和+11-05，三角形)组合时，与单独施用这些抗原引起的保护活性相比，4种抗原未改变保护的平均值。对于两种抗原，抗原组合(CSP+11-09和CSP+11-06)显示出更好的保护趋势(约10倍)，但不是统计学显著的。
5、鉴定能够通过CD8+T细胞应答对子孢子感染进行灭菌的多抗原制剂
由于检测所有可能的抗原组合在技术上是不可行的，因此发明人决定评估由这些多种保护性抗原的组合引起的保护作用。值得注意的是，使用CSP与6种这些抗原(10-05、11-06、11-07、11-09、11-10、18-10)的组合对小鼠进行的两次免疫，在绝大多数受到攻击的动物(7PEAg，86％至100％，图15)中引起了无菌保护。此种无菌保护百分比远优于CSP在相同实验条件下所赋予的保护作用(0-14％)。攻击前CD8+细胞(a-CD8)被耗竭而CD4+细胞不耗竭，将此种保护作用降低至CSP诱导的水平，表明这6种PE抗原的加入所引起的额外保护作用由CD8+T细胞介导。
仅使用7PEAg或7PEAg+30-03的单次免疫观察到相同的保护效力(图16a和16b，8PEAg)，以及8PEAg的无菌免疫对CD8+T细胞的依赖性(图16b)。由于抗原30-03是大分子并且当与7PEAg一起施用时没有改善无菌保护，发明人将其排除在进一步分析之外。
6、含有下选PE抗原保护结构域的嵌合抗原的设计
为了确定能够引发此种另外的保护性CD8+T细胞应答的最小抗原组合物，发明人首先鉴定了其保护活性依赖于这些T细胞的抗原。如图17b所示，在CD8+细胞耗竭后，使用TRAP、18-10和11-10的两次免疫诱导的保护作用显著降低。在CD8+细胞耗竭后，使用11-10的两次免疫所诱导的保护作用减少，但不是统计学显著的(图17b)。因此，发明人将CSP与CD8+依赖性保护性抗原TRAP、18-10和11-09分组，并分别加入11-10、11-107和11-06，以鉴定能像完整抗原组合一样无菌保护免疫动物的最小抗原组合。如图17a所示，5种抗原(CSP、TRAP、18-10、Ag40和Ag45)的组合诱导了与7PEAg的组合引起的无菌保护水平相当的无菌保护。
为将这5种抗原中每种的保护结构域组合在单个嵌合分子中，从而避免制备和递送五种不同抗原的相关成本，发明人根据结合MHC I类分子(图17c和18)和结构功能保守基序(图12，13，17c和18)的预测表位的定位，确定了每种抗原保护区的位置。
如图17c所示，与整个抗原相比，所有测试结构域呈现更好的(11-1OCT)或类似的保护活性。由诱导保护性CD8+T细胞的抗原结构域引发的平均保护水平与SYFPEITHI和IEDB(图17c和17d)分别预测的CD8+T细胞表位的得分(P<0.01)或亲和力(P＝0.01)相关。重要的是，保护结构域的定位(mapping)允许在最终的嵌合PE抗原构建体中减少约2000个碱基对。由于Ag40尺寸小，未将Ag40分成结构域，并且图17c中呈现的数据来自图11中所示的实验。
分析Pb抗原或Pf直系同源的表位分布，所述Pb抗原的表位被预测与C57BL/6小鼠的MHC I类分子(H2-Kb、H2-Db)结合，所述Pf直系同源的表位被预测与HLA A02:01(一种普遍存在的人类HLA等位基因)结合，揭示了大多数预测的良好结合物在不同疟原虫物种(Pb、Pc、Pv和Pf，图18)的保守区成簇(cluster)。这使得可以利用图17中标记的Pb保护区来选择Pf直系同源中的相应区域。此外，发明人使用Proimmune开发的结合方法验证了最佳的预测Pf表位与HLA A02:01I类分子的结合，其允许对由检测的肽(HLA A02:01和β2-微球蛋白)形成的复合物的结合和稳定化进行定量(图18)。
总之，使用抗原的参数化选择、基于慢病毒疫苗接种的筛选和针对严格的子孢子攻击的体内保护的直接测量，发明人从55种检测抗原中鉴定了8种保护性抗原，包括候选疫苗CSP和TRAP。所有这8种抗原在几种疟原虫物种中都是保守的。值得注意的是，使用七种或八种这些抗原的组合的免疫(使用一次或两次免疫)在绝大多数受攻击的小鼠中引发无菌保护。更重要的是，此种保护远优于CSP(迄今为止最好的保护性PE抗原)引发的保护。CD8+T细胞耗竭消除了无菌免疫，表明这些细胞对于此种保护性表型是必需的，类似于经辐照的子孢子赋予的保护作用。这些抗原中的5种的最小组合也能够在大多数受攻击的动物中引发无菌保护。对这5种抗原的保护结构域的定位，使得可以设计含有这5种下选抗原的融合保护结构域的嵌合抗原。这些保护性抗原或它们的保护结构域的人感染寄生虫直系同源是用于开发疟疾疫苗制剂的潜在候选物，其中该疟疾疫苗制剂含有融合在单个分子中的多个保护性抗原或多个保护性结构域。
材料与方法
寄生虫菌株：在hsp70启动子控制下，组成型表达GFP的伯氏疟原虫ANKA菌株(Ishino等，2006年)用于使用寄生虫血症的攻击，其通过流式细胞术定量作为保护性读数。在eef-1α启动子控制下，组成型表达GFP-荧光素酶融合体的伯氏疟原虫ANKA菌株(Franke-Fayard等，2008年)用于使用肝脏感染的攻击，其通过生物发光评估作为保护性读数。值得注意的是，因为GFP的表达水平更强，使用hsp70-gfp寄生虫的定量的寄生虫血症，比使用eef-1a gfp:luc寄生虫的灵敏度至少高10倍。
-Ishino T,Orito Y,Chinzei Y,Yuda M(2006)A calcium-dependent proteinkinase regulates Plasmodium ookinete access to the midgut epithelial cell.MolMicrobiol,59:1175-1184.
-Franke-Fayard B,Djokovic D,Dooren MW,Ramesar J,Waters AP,et al.(2008)Simple and sensitive antimalarial drug screening in vitro and in vivousing transgenic luciferase expressing Plasmodium berghei parasites.Int JParasitol,38:1651-1662.
小鼠品系：C57BL/6Rj和瑞士小鼠购自Elevage Janvier(法国)。所有实验均由巴斯德研究所动物护理和使用委员会(Animal Care and Use Committee of InstitutPasteur)(CETEA 2013-0093)批准，并且操作符合欧洲指南和法规(MESR-01324)。
慢病毒颗粒原液的生产：由Eurofins MWG合成下选疟原虫抗原作为小鼠密码子优化基因，并加有Kozak共有序列(GCCACCATGGCTC)(SEQ ID NO：85和86)，代表抗原多肽的编码序列中的前12个核苷酸，包括第一个翻译的ATG。该修饰在第一个甲硫氨酸后添加额外的丙氨酸。还将BamHI(GGATCC-SEQ ID NO：87)和Xho I(CTCGAG-SEQ ID NO：88)限制性位点分别插入构建体的5'和3'末端。然后将这些合成的密码子优化的基因克隆到含有CMV或B2M启动子的pTRIP质粒的BamHI和Xho I限制性位点中(图16和17)。通过用三个辅助质粒瞬时钙共转染HEK 293T细胞产生慢病毒颗粒，其中，三个辅助质粒编码单独的包装功能，pTRIP载体质粒含有合成的疟原虫基因，包膜表达质粒编码来自VSV(水疱性口炎病毒，印第安纳型(图19)或新泽西型(图20))的糖蛋白G，以及p8.74衣壳化质粒(图18)，含有HIV的结构、辅助和调节基因。此种共转染将产生整合但复制缺陷的假型慢病毒颗粒。在转染后24小时，用无血清DMEM更换细胞培养基。在转染后48小时收集上清液，通过低速离心澄清，并储存在-80℃。通过对来自转导的HEK293T细胞的细胞裂解物进行实时PCR来滴定慢病毒载体原液，滴度表示为转导单位(TU)/ml。
免疫方案：对于使用单剂量LP的筛选，将4周龄C57BL/6小鼠(每个实验每组n＝5)适应3周并且用单剂量1×107TU的非浓缩VSVIND B2M LP进行腹膜内免疫。对于使用两个免疫剂量的方案：使4周龄的C57BL/6小鼠(每个实验每组n＝5)适应3周，并用第一剂5×105TU的非浓缩VSVIND B2M LP进行腹膜内免疫；第一次免疫后30天，动物接受第二剂1×107TU的非浓缩VSVIND B2M LP。为了检测次优剂量的CSP+(最佳剂量的下选抗原)的组合，将小鼠免疫两次，间隔四周，用次优剂量的CSP(第一次免疫中的5×105TU的非浓缩VSVNJ B2M LP和第二次免疫中的5×106TU非浓缩VSVIND B2M LP)和通常剂量的保护性疟原虫抗原(第一次免疫中的5×105TU的非浓缩VSVNJ B2M LP和第二次免疫中的非浓缩1×107TU的VSVIND B2MLP)。为了检测多种抗原的组合，小鼠被免疫两次，间隔四周，7×个体剂量(1剂量＝第一次免疫中的5×105TU的非浓缩VSVNJ B2M LP/第二次免疫中的1×107TU的非浓缩VSVIND B2MLP)的对照抗原AI11荧光素酶(Luc)，以及个体剂量的CSP加6个剂量的Luc，或个体剂量的CSP和6种保守PE抗原(11-05、11-06、11-07、11-09、11-10和18-10)。对于该实验，发明人使用超滤和lenti-X(Clontech)浓缩原液。平均注射量为500μL用DMEM稀释的LP。
在所有情况下，在最后一次免疫后30天，用5,000个表达GFP的子孢子(皮下微量注射在小鼠足垫中)攻击小鼠。
子孢子攻击：使用标准方法饲养斯氏按蚊(Anopheles stephensi)(Sda500株)蚊子。羽化(emergence)后3至5天，用感染的瑞士小鼠(寄生虫血症高于2％)喂养该蚊子，并如Amino等(2007年)所述进行保存。在喂食后20至23天，将感染的蚊子的唾液腺在PBS中解剖，用20uL无菌PBS冰上收集，使用Eppendorf杵捣碎。通过40μm的尼龙网过滤寄生虫悬浮液，使用Kova Glasstic Slide载玻片(Hycor)计数，并用无菌PBS将浓度调节至5,000或10,000个子孢子/uL。将该悬浮液分成单独的管，每组免疫小鼠(每组n＝4-7)一个管，并保持在冰上直至攻击。使用具有35GA斜面针(NF35BV)的Nanofil注射器(WorldPrecisionInstruments)将1微升寄生虫悬浮液注射到小鼠的右足垫中。
Amino R,Thiberge S,Blazquez S,Baldacci P,Renaud O,et al.(2007)Imagingmalaria sporozoites in the dermis of the mammalian host.Nat Protoc 2:1705-1712.
寄生虫感染的测量：在约44h时，通过生物发光定量感染GFPLUC寄生虫的毛皮剃毛小鼠的肝寄生虫负荷。首先用异氟烷麻醉感染小鼠并皮下注射D-荧光素(150mg/kg，Caliper LifeSciences)。孵育5分钟，使基质在麻醉动物体内分布后，将小鼠转移至强化电荷耦合装置光子计数摄像机盒的阶段，在该阶段，通过鼻锥递送2.5％异氟烷维持麻醉状态。在Living Image软件(Xenogen Corporation)控制信号采集5分钟后，将动物放回笼中。系统自动检测生物发光信号从而产生叠加到实验小鼠的灰度照片上的生物发光信号图。然后使用Living Image软件量化这些图像。简言之，手动定义包含肝脏的目标区域(ROI)，应用于所有动物，并自动计算这些ROI内的平均辐射度。在腹部的下部区域测量背景信号，并获得背景信号的平均值。
或者，使用hsp70-GFP寄生虫通过流式细胞术评估血液感染。在攻击后第4、5、6和>10天，在小鼠尾部进行毫米切除，使得收集一滴血液，即时用500uL的PBS进行稀释。使用流式细胞仪分析该稀释的血液。攻击后第4天分析了500,000个事件，并在随后的几天分析了100,000个事件。第10天后未感染的小鼠被认为是受无菌保护的。
统计学分析：对来自GFP免疫对照的寄生虫血症数据进行log转换并合并以计算95％的容许区间，确信度(certitude)为95％。对于一个剂量的免疫方案，该限度包括平均值±2.49SD的区间(平均值＝-0.3906，SD＝0.3392，n＝35)。类似地，对于两个剂量的免疫方案，该限度包括平均值±2.51的区间(平均值＝-0.3002，SD＝0.3305，n＝33)。所有log(寄生虫血症)低于下限(平均值–2.5SD)的小鼠被认为与对照小鼠显著不同(P<0.05)，因此被认为是受保护的。在使用两个免疫剂量的方案中，根据上述定义，使用Fisher精确检验评估测试组和GFP对照组的受保护小鼠数量的差异。使用单向ANOVA(Holm-Sidak的多重比较检验)将具有Fisher检验中显著差异的组的log(寄生虫血症)的平均值值与GFP组进行比较。
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序列表
<110> 巴斯德研究所（INSTITUT PASTEUR）
法国国家科学研究中心（CENTRE DE LA RECHERCHE SCIENTIFIQUE）
<120> 多种疟疾红前期抗原及其在宿主中引发保护性免疫应答的用途
<130> B12096A-AD/BA
<140> PCT/EP2018/XXXXXX
<141> 2017-02-01
<150> 17305122.8
<151> 2017-02-02
<160> 94
<170> PatentIn version 3.5
<210> 1
<211> 4536
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> pTRIP CMV GFP
<400> 1
tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca 60
cacaaggcta cttccctgat tagcagaact acacaccagg gccagggatc agatatccac 120
tgacctttgg atggtgctac aagctagtac cagttgagcc agagaagtta gaagaagcca 180
acaaaggaga gaacaccagc ttgttacaac ctgtgagcct gcatgggatg gatgacccgg 240
agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac ggtggcccga 300
gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc 360
gctgggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag 420
atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct 480
gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc 540
cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc 600
tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa 660
agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac 720
ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta 780
gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg 840
ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg 900
ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg 960
ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag 1020
atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga 1080
caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca 1140
gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg 1200
aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa 1260
agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt 1320
tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca 1380
gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc 1440
aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg 1500
ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac 1560
tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga 1620
tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa 1680
tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg 1740
aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata 1800
taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac 1860
tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc 1920
caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca 1980
gagacagatc cattcgatta gtgaacggat ctcgacggta tcgccgaatt cacaaatggc 2040
agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag 2100
aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa 2160
aattcaaaat tttcgggttt attacaggga cagcagagat ccactttggc tgatacgcgt 2220
ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc 2280
ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca 2340
ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta 2400
tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta 2460
tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat 2520
cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga 2580
ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca 2640
aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg 2700
taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc 2760
ctggagacgc catccacgct gttttgacct ccatagaaga caccgcgatc ggatccccac 2820
cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg 2880
tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg 2940
atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc 3000
cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg 3060
accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc 3120
gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg 3180
gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca 3240
tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca 3300
agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg 3360
tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc 3420
ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg 3480
atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc 3540
tgtacaagta aagcggccgc gactctagct cgagctcaag cttcgaattc ccgataatca 3600
acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt 3660
tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc 3720
tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc 3780
cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg 3840
gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc 3900
cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg 3960
cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg 4020
tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc 4080
agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct 4140
tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcggg aattctgcag 4200
tcgacggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta 4260
aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tcgtcgagag 4320
atgctgcata taagcagctg ctttttgctt gtactgggtc tctctggtta gaccagatct 4380
gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc 4440
cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc 4500
tcagaccctt ttagtcagtg tggaaaatct ctagca 4536
<210> 2
<211> 4468
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> pTRIP B2M GFP
<400> 2
tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca 60
cacaaggcta cttccctgat tagcagaact acacaccagg gccagggatc agatatccac 120
tgacctttgg atggtgctac aagctagtac cagttgagcc agagaagtta gaagaagcca 180
acaaaggaga gaacaccagc ttgttacaac ctgtgagcct gcatgggatg gatgacccgg 240
agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac ggtggcccga 300
gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc 360
gctgggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag 420
atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct 480
gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc 540
cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc 600
tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa 660
agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac 720
ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta 780
gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg 840
ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg 900
ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg 960
ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag 1020
atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga 1080
caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca 1140
gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg 1200
aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa 1260
agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt 1320
tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca 1380
gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc 1440
aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg 1500
ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac 1560
tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga 1620
tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa 1680
tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg 1740
aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata 1800
taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac 1860
tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc 1920
caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca 1980
gagacagatc cattcgatta gtgaacggat ctcgacggta tcgccgaatt cacaaatggc 2040
agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag 2100
aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa 2160
aattcaaaat tttcgggttt attacaggga cagcagagat ccactttggc tgatacgcgc 2220
cggaaaccct gcagggaatt ccccagctgt agttataaac agaagttctc cttctgctag 2280
gtagcattca aagatcttaa tcttctgggt ttccgttttc tcgaatgaaa aatgcaggtc 2340
cgagcagtta actggcgggg gcaccattag caagtcactt agcatctctg gggccagtct 2400
gcaaagcgag ggggcagcct taatgtgcct ccagcctgaa gtcctagaat gagcgcccgg 2460
tgtcccaagc tggggcgcgc accccagatc ggagggcgcc gatgtacaga cagcaaactc 2520
acccagtcta gtgcatgcct tcttaaacat cacgagactc taagaaaagg aaactgaaaa 2580
cgggaaagtc cctctctcta acctggcact gcgtcgctgg cttggagaca ggtgacggtc 2640
cctgcgggcc ttgtcctgat tggctgggca cgcgtttaat ataagtggag gcgtcgcgct 2700
ggcgggcatt cctgaagctg acagcattcg ggccgagcga tcggatcccc accggtcgcc 2760
accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg 2820
gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc 2880
tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc 2940
accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg 3000
aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc 3060
ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc 3120
ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg 3180
cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag 3240
aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc 3300
gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac 3360
cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg 3420
gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag 3480
taaagcggcc gcgactctag ctcgagctca agcttcgaat tcccgataat caacctctgg 3540
attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat 3600
gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt 3660
tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca 3720
ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg 3780
ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg 3840
aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca 3900
attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca 3960
cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc 4020
ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc 4080
agacgagtcg gatctccctt tgggccgcct ccccgcatcg ggaattctgc agtcgacggt 4140
acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa 4200
ggggggactg gaagggctaa ttcactccca acgaagacaa gatcgtcgag agatgctgca 4260
tataagcagc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg 4320
gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg 4380
cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc 4440
ttttagtcag tgtggaaaat ctctagca 4468
<210> 3
<211> 11904
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 包装8.74质粒
<400> 3
gggctgcagg aattaattcg agctcgcccg acattgatta ttgactagtt attaatagta 60
atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac 120
ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac 180
gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt 240
acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat 300
tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga 360
ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt 420
ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca 480
ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg 540
tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta 600
tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt 660
tgacctccat agaagacacc gggaccgatc cagcctccgc ggccgcgttg acgcgcacgg 720
caagaggcga ggggcggcga ctggtgagag atgggtgcga gagcgtcagt attaagcggg 780
ggagaattag atcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa 840
ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg 900
ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca 960
ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa 1020
aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa 1080
agtaagaaaa aagcacagca agcagcagct gacacaggac acagcaatca ggtcagccaa 1140
aattacccta tagtgcagaa catccagggg caaatggtac atcaggccat atcacctaga 1200
actttaaatg catgggtaaa agtagtagaa gagaaggctt tcagcccaga agtgataccc 1260
atgttttcag cattatcaga aggagccacc ccacaagatt taaacaccat gctaaacaca 1320
gtggggggac atcaagcagc catgcaaatg ttaaaagaga ccatcaatga ggaagctgca 1380
gaatgggata gagtgcatcc agtgcatgca gggcctattg caccaggcca gatgagagaa 1440
ccaaggggaa gtgacatagc aggaactact agtacccttc aggaacaaat aggatggatg 1500
acacataatc cacctatccc agtaggagaa atctataaaa gatggataat cctgggatta 1560
aataaaatag taagaatgta tagccctacc agcattctgg acataagaca aggaccaaag 1620
gaacccttta gagactatgt agaccgattc tataaaactc taagagccga gcaagcttca 1680
caagaggtaa aaaattggat gacagaaacc ttgttggtcc aaaatgcgaa cccagattgt 1740
aagactattt taaaagcatt gggaccagga gcgacactag aagaaatgat gacagcatgt 1800
cagggagtgg ggggacccgg ccataaagca agagttttgg ctgaagcaat gagccaagta 1860
acaaatccag ctaccataat gatacagaaa ggcaatttta ggaaccaaag aaagactgtt 1920
aagtgtttca attgtggcaa agaagggcac atagccaaaa attgcagggc ccctaggaaa 1980
aagggctgtt ggaaatgtgg aaaggaagga caccaaatga aagattgtac tgagagacag 2040
gctaattttt tagggaagat ctggccttcc cacaagggaa ggccagggaa ttttcttcag 2100
agcagaccag agccaacagc cccaccagaa gagagcttca ggtttgggga agagacaaca 2160
actccctctc agaagcagga gccgatagac aaggaactgt atcctttagc ttccctcaga 2220
tcactctttg gcagcgaccc ctcgtcacaa taaagatagg ggggcaatta aaggaagctc 2280
tattagatac aggagcagat gatacagtat tagaagaaat gaatttgcca ggaagatgga 2340
aaccaaaaat gataggggga attggaggtt ttatcaaagt aagacagtat gatcagatac 2400
tcatagaaat ctgcggacat aaagctatag gtacagtatt agtaggacct acacctgtca 2460
acataattgg aagaaatctg ttgactcaga ttggctgcac tttaaatttt cccattagtc 2520
ctattgagac tgtaccagta aaattaaagc caggaatgga tggcccaaaa gttaaacaat 2580
ggccattgac agaagaaaaa ataaaagcat tagtagaaat ttgtacagaa atggaaaagg 2640
aaggaaaaat ttcaaaaatt gggcctgaaa atccatacaa tactccagta tttgccataa 2700
agaaaaaaga cagtactaaa tggagaaaat tagtagattt cagagaactt aataagagaa 2760
ctcaagattt ctgggaagtt caattaggaa taccacatcc tgcagggtta aaacagaaaa 2820
aatcagtaac agtactggat gtgggcgatg catatttttc agttccctta gataaagact 2880
tcaggaagta tactgcattt accataccta gtataaacaa tgagacacca gggattagat 2940
atcagtacaa tgtgcttcca cagggatgga aaggatcacc agcaatattc cagtgtagca 3000
tgacaaaaat cttagagcct tttagaaaac aaaatccaga catagtcatc tatcaataca 3060
tggatgattt gtatgtagga tctgacttag aaatagggca gcatagaaca aaaatagagg 3120
aactgagaca acatctgttg aggtggggat ttaccacacc agacaaaaaa catcagaaag 3180
aacctccatt cctttggatg ggttatgaac tccatcctga taaatggaca gtacagccta 3240
tagtgctgcc agaaaaggac agctggactg tcaatgacat acagaaatta gtgggaaaat 3300
tgaattgggc aagtcagatt tatgcaggga ttaaagtaag gcaattatgt aaacttctta 3360
ggggaaccaa agcactaaca gaagtagtac cactaacaga agaagcagag ctagaactgg 3420
cagaaaacag ggagattcta aaagaaccgg tacatggagt gtattatgac ccatcaaaag 3480
acttaatagc agaaatacag aagcaggggc aaggccaatg gacatatcaa atttatcaag 3540
agccatttaa aaatctgaaa acaggaaagt atgcaagaat gaagggtgcc cacactaatg 3600
atgtgaaaca attaacagag gcagtacaaa aaatagccac agaaagcata gtaatatggg 3660
gaaagactcc taaatttaaa ttacccatac aaaaggaaac atgggaagca tggtggacag 3720
agtattggca agccacctgg attcctgagt gggagtttgt caatacccct cccttagtga 3780
agttatggta ccagttagag aaagaaccca taataggagc agaaactttc tatgtagatg 3840
gggcagccaa tagggaaact aaattaggaa aagcaggata tgtaactgac agaggaagac 3900
aaaaagttgt ccccctaacg gacacaacaa atcagaagac tgagttacaa gcaattcatc 3960
tagctttgca ggattcggga ttagaagtaa acatagtgac agactcacaa tatgcattgg 4020
gaatcattca agcacaacca gataagagtg aatcagagtt agtcagtcaa ataatagagc 4080
agttaataaa aaaggaaaaa gtctacctgg catgggtacc agcacacaaa ggaattggag 4140
gaaatgaaca agtagataaa ttggtcagtg ctggaatcag gaaagtacta tttttagatg 4200
gaatagataa ggcccaagaa gaacatgaga aatatcacag taattggaga gcaatggcta 4260
gtgattttaa cctaccacct gtagtagcaa aagaaatagt agccagctgt gataaatgtc 4320
agctaaaagg ggaagccatg catggacaag tagactgtag cccaggaata tggcagctag 4380
attgtacaca tttagaagga aaagttatct tggtagcagt tcatgtagcc agtggatata 4440
tagaagcaga agtaattcca gcagagacag ggcaagaaac agcatacttc ctcttaaaat 4500
tagcaggaag atggccagta aaaacagtac atacagacaa tggcagcaat ttcaccagta 4560
ctacagttaa ggccgcctgt tggtgggcgg ggatcaagca ggaatttggc attccctaca 4620
atccccaaag tcaaggagta atagaatcta tgaataaaga attaaagaaa attataggac 4680
aggtaagaga tcaggctgaa catcttaaga cagcagtaca aatggcagta ttcatccaca 4740
attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa 4800
tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc 4860
gggtttatta cagggacagc agagatccag tttggaaagg accagcaaag ctcctctgga 4920
aaggtgaagg ggcagtagta atacaagata atagtgacat aaaagtagtg ccaagaagaa 4980
aagcaaagat catcagggat tatggaaaac agatggcagg tgatgattgt gtggcaagta 5040
gacaggatga ggattaacac atggaattct gcaacaactg ctgtttatcc atttcagaat 5100
tgggtgtcga catagcagaa taggcgttac tcgacagagg agagcaagaa atggagccag 5160
tagatcctag actagagccc tggaagcatc caggaagtca gcctaaaact gcttgtacca 5220
attgctattg taaaaagtgt tgctttcatt gccaagtttg tttcatgaca aaagccttag 5280
gcatctccta tggcaggaag aagcggagac agcgacgaag agctcatcag aacagtcaga 5340
ctcatcaagc ttctctatca aagcagtaag tagtacatgt aatgcaacct ataatagtag 5400
caatagtagc attagtagta gcaataataa tagcaatagt tgtgtggtcc atagtaatca 5460
tagaatatag gaaaatggcc gctgatcttc agacctggag gaggagatat gagggacaat 5520
tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc 5580
accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg 5640
ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg 5700
gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct 5760
attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca 5820
agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc 5880
tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct 5940
ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac 6000
acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa 6060
gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg 6120
ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt 6180
tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag 6240
acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga 6300
gagagagaca gagacagatc cattcgatta gtgaacggat ccttggcact tatctgggac 6360
gatctgcgga gcctgtgcct cttcagctac caccgcttga gagacttact cttgattgta 6420
acgaggattg tggaacttct gggacgcagg gggtgggaag ccctcaaata ttggtggaat 6480
ctcctacaat attggagtca ggagctaaag aatagtgctg ttagcttgct caatgccaca 6540
gccatagcag tagctgaggg gacagatagg gttatagaag tagtacaagg agcttgtaga 6600
gctattcgcc acatacctag aagaataaga cagggcttgg aaaggatttt gctataagct 6660
cgagtgacct tcagaccttg gcactggagg tggcccggca gaagcgcggc atcgtggatc 6720
agtgctgcac cagcatctgc tctctctacc aactggagaa ctactgcaac taggcccacc 6780
actaccctgt ccacccctct gcaatgaata aaacctttga aagagcacta caagttgtgt 6840
gtacatgcgt gcatgtgcat atgtggtgcg gggggaacat gagtggggct ggctggagtg 6900
gcgatgataa gctgtcaaac atgagaatta attcttgaag acgaaagggc ctcgtgatac 6960
gcctattttt ataggttaat gtcatgataa taatggtttc ttagtctaga attaattccg 7020
tgtattctat agtgtcacct aaatcgtatg tgtatgatac ataaggttat gtattaattg 7080
tagccgcgtt ctaacgacaa tatgtacaag cctaattgtg tagcatctgg cttactgaag 7140
cagaccctat catctctctc gtaaactgcc gtcagagtcg gtttggttgg acgaaccttc 7200
tgagtttctg gtaacgccgt cccgcacccg gaaatggtca gcgaaccaat cagcagggtc 7260
atcgctagcc agatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt 7320
gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc 7380
gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg 7440
ttgggcgcca tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac 7500
ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgtcg aatggtgcac 7560
tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc 7620
cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac 7680
cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg 7740
aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta 7800
gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta 7860
aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata 7920
ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc 7980
ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga 8040
agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct 8100
tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg 8160
tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta 8220
ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat 8280
gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt 8340
acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga 8400
tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga 8460
gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga 8520
actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc 8580
aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc 8640
cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg 8700
tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat 8760
cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata 8820
tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct 8880
ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga 8940
ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 9000
cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 9060
aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct 9120
agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 9180
tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 9240
ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 9300
cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct 9360
atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 9420
ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 9480
tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 9540
gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 9600
gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac 9660
cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt 9720
gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat 9780
tcattaatgc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca 9840
gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc 9900
ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata 9960
gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 10020
ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag 10080
ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttgga 10140
cacaagacag gcttgcgaga tatgtttgag aataccactt tatcccgcgt cagggagagg 10200
cagtgcgtaa aaagacgcgg actcatgtga aatactggtt tttagtgcgc cagatctcta 10260
taatctcgcg caacctattt tcccctcgaa cactttttaa gccgtagata aacaggctgg 10320
gacacttcac atgagcgaaa aatacatcgt cacctgggac atgttgcaga tccatgcacg 10380
taaactcgca agccgactga tgccttctga acaatggaaa ggcattattg ccgtaagccg 10440
tggcggtctg taccgggtgc gttactggcg cgtgaactgg gtattcgtca tgtcgatacc 10500
gtttgtattt ccagctacga tcacgacaac cagcgcgagc ttaaagtgct gaaacgcgca 10560
gaaggcgatg gcgaaggctt catcgttatt gatgacctgg tggataccgg tggtactgcg 10620
gttgcgattc gtgaaatgta tccaaaagcg cactttgtca ccatcttcgc aaaaccggct 10680
ggtcgtccgc tggttgatga ctatgttgtt gatatcccgc aagatacctg gattgaacag 10740
ccgtgggata tgggcgtcgt attcgtcccg ccaatctccg gtcgctaatc ttttcaacgc 10800
ctggcactgc cgggcgttgt tctttttaac ttcaggcggg ttacaatagt ttccagtaag 10860
tattctggag gctgcatcca tgacacaggc aaacctgagc gaaaccctgt tcaaaccccg 10920
ctttaaacat cctgaaacct cgacgctagt ccgccgcttt aatcacggcg cacaaccgcc 10980
tgtgcagtcg gcccttgatg gtaaaaccat ccctcactgg tatcgcatga ttaaccgtct 11040
gatgtggatc tggcgcggca ttgacccacg cgaaatcctc gacgtccagg cacgtattgt 11100
gatgagcgat gccgaacgta ccgacgatga tttatacgat acggtgattg gctaccgtgg 11160
cggcaactgg atttatgagt gggccccgga tctttgtgaa ggaaccttac ttctgtggtg 11220
tgacataatt ggacaaacta cctacagaga tttaaagctc taaggtaaat ataaaatttt 11280
taagtgtata atgtgttaaa ctactgattc taattgtttg tgtattttag attccaacct 11340
atggaactga tgaatgggag cagtggtgga atgcctttaa tgaggaaaac ctgttttgct 11400
cagaagaaat gccatctagt gatgatgagg ctactgctga ctctcaacat tctactcctc 11460
caaaaaagaa gagaaaggta gaagacccca aggactttcc ttcagaattg ctaagttttt 11520
tgagtcatgc tgtgtttagt aatagaactc ttgcttgctt tgctatttac accacaaagg 11580
aaaaagctgc actgctatac aagaaaatta tggaaaaata ttctgtaacc tttataagta 11640
ggcataacag ttataatcat aacatactgt tttttcttac tccacacagg catagagtgt 11700
ctgctattaa taactatgct caaaaattgt gtacctttag ctttttaatt tgtaaagggg 11760
ttaataagga atatttgatg tatagtgcct tgactagaga tcataatcag ccataccaca 11820
tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat 11880
aaaatgaatg caattgttgt tgtt 11904
<210> 4
<211> 5140
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> pCMV-VSV-INDco
<400> 4
ctggatggct ttctcgccgc caaggatctg atggcgcagg ggatcaagct ctgatcaaga 60
gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc 120
cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga 180
tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct 240
gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac 300
gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct 360
attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt 420
atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt 480
cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt 540
cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag 600
gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt 660
gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg 720
tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg 780
cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg 840
catcgccttc tatcgccttc ttgacgagtt cttctgaatt attaacgctt acaatttcct 900
gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac aggtggcact 960
tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg 1020
tatccgctca tgagacaata accctgataa atgcttcaat aatagcacgt gctaaaactt 1080
catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc 1140
ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 1200
tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 1260
ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 1320
ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac 1380
ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 1440
gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 1500
aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 1560
acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 1620
gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 1680
gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 1740
cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc 1800
aacgcggcct ttttacggtt cctgggcttt tgctggcctt ttgctcacat gttcttgact 1860
cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag 1920
taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt 1980
acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg 2040
acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggactat 2100
ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct 2160
attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg 2220
gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg 2280
ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc 2340
caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa 2400
tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc 2460
tatataagca gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt 2520
aatacgactc actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg 2580
agctcggatc ctgatcagcc accatgaaat gcctgctcta tctggccttc ctgttcatcg 2640
gcgtgaactg caagttcacc atcgtgttcc cccacaacca gaagggcaac tggaagaacg 2700
tgcccagcaa ctaccactac tgccccagca gcagcgacct gaactggcac aacgacctga 2760
tcggcaccgc cctgcaggtg aagatgccca agagccacaa ggccattcag gctgatggct 2820
ggatgtgtca tgccagcaag tgggtgacca cctgcgactt ccggtggtac ggccccaagt 2880
acatcaccca cagcatccgc agcttcaccc ccagcgtgga gcagtgcaaa gagagcatcg 2940
agcagaccaa gcagggcacc tggctgaacc ccggcttccc cccccagtcc tgcggctacg 3000
ccaccgtgac cgacgccgag gccgtgatcg tgcaggtgac cccccaccac gtgctggtcg 3060
acgagtacac cggcgagtgg gtggacagcc agttcatcaa cggcaagtgc agcaactaca 3120
tctgccctac cgtgcacaac agcaccacct ggcacagcga ctacaaggtg aaaggcctgt 3180
gcgacagcaa cctgatcagc atggacatca cctttttcag cgaggacggc gagctgtcca 3240
gcctgggcaa agagggcacc ggcttcagaa gcaactactt cgcctacgag acaggcggca 3300
aggcctgcaa gatgcagtac tgcaagcact ggggcgtgcg gctgcctagc ggcgtgtggt 3360
tcgagatggc cgacaaggac ctgttcgccg ctgcccggtt ccctgagtgc cccgagggca 3420
gcagcatcag cgcccccagc cagaccagcg tggacgtgag cctgatccag gacgtggaga 3480
gaatcctgga ctacagcctg tgccaggaaa cctggtccaa gatcagagcc ggcctgccca 3540
tcagccccgt ggacctgagc tacctggccc ccaagaaccc cggcaccggc ccagccttca 3600
ccatcatcaa tggcaccctg aagtacttcg agacacggta catcagagtg gacattgccg 3660
cccctatcct gagccggatg gtgggcatga tcagcggcac caccaccgag cgggagctgt 3720
gggacgactg ggccccctac gaggatgtgg agatcggccc caacggcgtg ctgcggacca 3780
gcagcggcta caagttcccc ctgtacatga tcggccacgg catgctggac agcgacctgc 3840
acctgagcag caaggcccag gtgttcgagc acccccacat ccaggacgcc gccagccagc 3900
tgcccgacga cgagagcctg ttcttcggcg acaccggcct gagcaagaac cccatcgaac 3960
tggtggaggg ctggttcagc agctggaaga gcagcattgc cagctttttc ttcatcatcg 4020
gcctgatcat cgggctgttt ctggtgctga gagtgggcat ccacctgtgc atcaagctga 4080
agcacaccaa gaagcggcag atctacaccg acatcgagat gaatcgcctg gggaagtaag 4140
aattctgcag atatccagca cagtggcggc cgctcgagtg tacaaattcc cgataatcaa 4200
cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt 4260
acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct 4320
ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc 4380
gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg 4440
ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc 4500
acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc 4560
actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatcgct gctcgcctgt 4620
gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca 4680
gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt 4740
cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgctcgagtc tagagggccc 4800
gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc 4860
ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa 4920
aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg 4980
gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg 5040
ggctctatgg cttctactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg 5100
gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa 5140
<210> 5
<211> 5158
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> pCMV-VSV-Njco
<400> 5
ctggatggct ttctcgccgc caaggatctg atggcgcagg ggatcaagct ctgatcaaga 60
gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc 120
cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga 180
tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct 240
gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac 300
gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct 360
attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt 420
atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt 480
cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt 540
cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag 600
gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt 660
gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg 720
tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg 780
cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg 840
catcgccttc tatcgccttc ttgacgagtt cttctgaatt attaacgctt acaatttcct 900
gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac aggtggcact 960
tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg 1020
tatccgctca tgagacaata accctgataa atgcttcaat aatagcacgt gctaaaactt 1080
catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc 1140
ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 1200
tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 1260
ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 1320
ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac 1380
ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 1440
gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 1500
aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 1560
acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 1620
gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 1680
gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 1740
cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc 1800
aacgcggcct ttttacggtt cctgggcttt tgctggcctt ttgctcacat gttcttgact 1860
cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag 1920
taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt 1980
acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg 2040
acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggactat 2100
ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct 2160
attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg 2220
gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg 2280
ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc 2340
caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa 2400
tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc 2460
tatataagca gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt 2520
aatacgactc actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg 2580
agctcggatc ctgatcagcc accatgctgt catatctgat ctttgccctg gctgtgagcc 2640
caatcctcgg aaagatcgaa atcgtgttcc cacaacacac cacaggggac tggaagcgcg 2700
tgccccacga gtacaactac tgcccgacct cagccgacaa gaatagccac ggcacgcaga 2760
ccggcatccc tgtggagctg accatgccca aggggctcac aacgcaccaa gtcgaaggct 2820
tcatgtgcca cagcgctctc tggatgacaa cctgcgattt tcgctggtat ggccccaagt 2880
acatcacgca cagcatccac aatgaggaac caaccgacta ccagtgcctc gaagccatca 2940
agtcatacaa ggatggggtg agcttcaacc ccggcttccc gccccaatca tgtggctacg 3000
gcaccgtgac cgacgccgag gcccacatcg tgaccgtgac accccactca gtcaaggtgg 3060
acgagtacac aggcgaatgg atcgaccccc acttcatcgg gggccgctgt aagggccaaa 3120
tctgcgagac cgtgcacaac agcaccaagt ggtttacgtc atcagacggc gaaagcgtgt 3180
gcagccaact gtttacgctc gtgggcggca tcttctttag cgacagcgag gagatcacca 3240
gcatgggcct cccggagaca ggaatccgca gcaactactt tccgtacatc agcaccgagg 3300
gaatctgtaa gatgcctttt tgccgcaagc agggatataa gctgaagaat gacctgtggt 3360
tccagatcat ggacccggac ctggacaaga ccgtccgcga tctgccccac atcaaggact 3420
gtgatctgtc atcaagcatc atcacccccg gagaacacgc cacggacatc agcctcatca 3480
gcgatgtgga gcgcatcctc gactacgctc tctgccagaa cacatggagc aagatcgaaa 3540
gcggcgaacc catcacccca gtggacctga gctatctcgg cccaaagaac cccggcgtgg 3600
ggcccgtgtt caccatcatc aacgggagcc tgcactactt tacaagcaag tatctgcgcg 3660
tggagctcga aagcccagtc atcccccgca tggaggggaa ggtggccggg acccgcatcg 3720
tgcgccagct gtgggaccag tggttccctt ttggcgaggt ggaaatcggc cccaacggcg 3780
tgctgaagac caagcaagga tataagttcc cgctgcacat catcgggacg ggcgaagtgg 3840
acagcgatat caagatggag cgcgtggtca agcactggga gcacccacac atcgaggctg 3900
ctcagacctt tctcaagaag gacgataccg gcgaagtcct gtattacggg gatacgggag 3960
tgagcaagaa ccctgtggag ctggtggaag gctggttcag cggatggcgc tcaagcctga 4020
tgggcgtgct ggccgtcatc atcggatttg tgatcctgat gttcctcatc aagctgatcg 4080
gcgtgctgtc aagcctgttc cgccctaagc gccgcccaat ctacaagagc gacgtcgaga 4140
tggcccactt tcgctaagaa ttctgcagat atccagcaca gtggcggccg ctcgagtgta 4200
caaattcccg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt 4260
aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct 4320
attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt 4380
tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac 4440
gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct 4500
ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca 4560
ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt 4620
ccatcgctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc 4680
ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct 4740
cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg 4800
ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc 4860
cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc 4920
actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct 4980
attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg 5040
catgctgggg atgcggtggg ctctatggct tctactgggc ggttttatgg acagcaagcg 5100
aaccggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc aaagtaaa 5158
<210> 6
<211> 720
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> eGFP
<400> 6
atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac 60
ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac 120
ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc 180
ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag 240
cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc 300
ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg 360
gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac 420
aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac 480
ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc 540
gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac 600
tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc 660
ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa 720
<210> 7
<211> 239
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> eGFP蛋白
<400> 7
Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu
1 5 10 15
Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly
20 25 30
Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile
35 40 45
Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr
50 55 60
Leu Thr Tyr Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys
65 70 75 80
Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu
85 90 95
Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu
100 105 110
Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly
115 120 125
Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr
130 135 140
Asn Tyr Asn Ser His Asn Val Tyr Ile Met Ala Asp Lys Gln Lys Asn
145 150 155 160
Gly Ile Lys Val Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser
165 170 175
Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly
180 185 190
Pro Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu
195 200 205
Ser Lys Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe
210 215 220
Val Thr Ala Ala Gly Ile Thr Leu Gly Met Asp Glu Leu Tyr Lys
225 230 235
<210> 8
<211> 1725
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> A111-荧光素酶
<400> 8
ggatccgcca ccatggccgc agccgcagcc gtgaagaact ggatgactca aacgctcgcc 60
gctgcaatgg aagatgccaa aaacattaag aagggcccag cgccattcta cccactcgaa 120
gacgggaccg ccggcgagca gctgcacaaa gccatgaagc gctacgccct ggtgcccggc 180
accatcgcct ttaccgacgc acatatcgag gtggacatta cctacgccga gtacttcgag 240
atgagcgttc ggctggcaga agctatgaag cgctatgggc tgaatacaaa ccatcggatc 300
gtggtgtgca gcgagaatag cttgcagttc ttcatgcccg tgttgggtgc cctgttcatc 360
ggtgtggctg tggccccagc taacgacatc tacaacgagc gcgagctgct gaacagcatg 420
ggcatcagcc agcccaccgt cgtattcgtg agcaagaaag ggctgcaaaa gatcctcaac 480
gtgcaaaaga agctaccgat catacaaaag atcatcatca tggatagcaa gaccgactac 540
cagggcttcc aaagcatgta caccttcgtg acttcccatt tgccacccgg cttcaacgag 600
tacgacttcg tgcccgagag cttcgaccgg gacaaaacca tcgccctgat catgaacagt 660
agtggcagta ccggattgcc caagggcgta gccctaccgc accgcaccgc ttgtgtccga 720
ttcagtcatg cccgcgaccc catcttcggc aaccagatca tccccgacac cgctatcctc 780
agcgtggtgc catttcacca cggcttcggc atgttcacca cgctgggcta cttgatctgc 840
ggctttcggg tcgtgctcat gtaccgcttc gaggaggagc tattcttgcg cagcttgcaa 900
gactataaga ttcaatctgc cctgctggtg cccacactat ttagcttctt cgctaagagc 960
actctcatcg acaagtacga cctaagcaac ttgcacgaga tcgccagcgg cggggcgccg 1020
ctcagcaagg aggtaggtga ggccgtggcc aaacgcttcc acctaccagg catccgccag 1080
ggctacggcc tgacagaaac aaccagcgcc attctgatca cccccgaagg ggacgacaag 1140
cctggcgcag taggcaaggt ggtgcccttc ttcgaggcta aggtggtgga cttggacacc 1200
ggtaagacac tgggtgtgaa ccagcgcggc gagctgtgcg tccgtggccc catgatcatg 1260
agcggctacg ttaacaaccc cgaggctaca aacgctctca tcgacaagga cggctggctg 1320
cacagcggcg acatcgccta ctgggacgag gacgagcact tcttcatcgt ggaccggctg 1380
aagagcctga tcaaatacaa gggctaccag gtagccccag ccgaactgga gagcatcctg 1440
ctgcaacacc ccaacatctt cgacgccggg gtcgccggcc tgcccgacga cgatgccggc 1500
gagctgcccg ccgcagtcgt cgtgctggaa cacggtaaaa ccatgaccga gaaggagatc 1560
gtggactatg tggccagcca ggttacaacc gccaagaagc tgcgcggtgg tgttgtgttc 1620
gtggacgagg tgcctaaagg actgaccggc aagttggacg cccgcaagat ccgcgagatt 1680
ctcattaagg ccaagaaggg cggcaagatc gccgtgtaac tcgag 1725
<210> 9
<211> 568
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> A111-荧光素酶
<400> 9
Met Ala Ala Ala Ala Ala Val Lys Asn Trp Met Thr Gln Thr Leu Ala
1 5 10 15
Ala Ala Met Glu Asp Ala Lys Asn Ile Lys Lys Gly Pro Ala Pro Phe
20 25 30
Tyr Pro Leu Glu Asp Gly Thr Ala Gly Glu Gln Leu His Lys Ala Met
35 40 45
Lys Arg Tyr Ala Leu Val Pro Gly Thr Ile Ala Phe Thr Asp Ala His
50 55 60
Ile Glu Val Asp Ile Thr Tyr Ala Glu Tyr Phe Glu Met Ser Val Arg
65 70 75 80
Leu Ala Glu Ala Met Lys Arg Tyr Gly Leu Asn Thr Asn His Arg Ile
85 90 95
Val Val Cys Ser Glu Asn Ser Leu Gln Phe Phe Met Pro Val Leu Gly
100 105 110
Ala Leu Phe Ile Gly Val Ala Val Ala Pro Ala Asn Asp Ile Tyr Asn
115 120 125
Glu Arg Glu Leu Leu Asn Ser Met Gly Ile Ser Gln Pro Thr Val Val
130 135 140
Phe Val Ser Lys Lys Gly Leu Gln Lys Ile Leu Asn Val Gln Lys Lys
145 150 155 160
Leu Pro Ile Ile Gln Lys Ile Ile Ile Met Asp Ser Lys Thr Asp Tyr
165 170 175
Gln Gly Phe Gln Ser Met Tyr Thr Phe Val Thr Ser His Leu Pro Pro
180 185 190
Gly Phe Asn Glu Tyr Asp Phe Val Pro Glu Ser Phe Asp Arg Asp Lys
195 200 205
Thr Ile Ala Leu Ile Met Asn Ser Ser Gly Ser Thr Gly Leu Pro Lys
210 215 220
Gly Val Ala Leu Pro His Arg Thr Ala Cys Val Arg Phe Ser His Ala
225 230 235 240
Arg Asp Pro Ile Phe Gly Asn Gln Ile Ile Pro Asp Thr Ala Ile Leu
245 250 255
Ser Val Val Pro Phe His His Gly Phe Gly Met Phe Thr Thr Leu Gly
260 265 270
Tyr Leu Ile Cys Gly Phe Arg Val Val Leu Met Tyr Arg Phe Glu Glu
275 280 285
Glu Leu Phe Leu Arg Ser Leu Gln Asp Tyr Lys Ile Gln Ser Ala Leu
290 295 300
Leu Val Pro Thr Leu Phe Ser Phe Phe Ala Lys Ser Thr Leu Ile Asp
305 310 315 320
Lys Tyr Asp Leu Ser Asn Leu His Glu Ile Ala Ser Gly Gly Ala Pro
325 330 335
Leu Ser Lys Glu Val Gly Glu Ala Val Ala Lys Arg Phe His Leu Pro
340 345 350
Gly Ile Arg Gln Gly Tyr Gly Leu Thr Glu Thr Thr Ser Ala Ile Leu
355 360 365
Ile Thr Pro Glu Gly Asp Asp Lys Pro Gly Ala Val Gly Lys Val Val
370 375 380
Pro Phe Phe Glu Ala Lys Val Val Asp Leu Asp Thr Gly Lys Thr Leu
385 390 395 400
Gly Val Asn Gln Arg Gly Glu Leu Cys Val Arg Gly Pro Met Ile Met
405 410 415
Ser Gly Tyr Val Asn Asn Pro Glu Ala Thr Asn Ala Leu Ile Asp Lys
420 425 430
Asp Gly Trp Leu His Ser Gly Asp Ile Ala Tyr Trp Asp Glu Asp Glu
435 440 445
His Phe Phe Ile Val Asp Arg Leu Lys Ser Leu Ile Lys Tyr Lys Gly
450 455 460
Tyr Gln Val Ala Pro Ala Glu Leu Glu Ser Ile Leu Leu Gln His Pro
465 470 475 480
Asn Ile Phe Asp Ala Gly Val Ala Gly Leu Pro Asp Asp Asp Ala Gly
485 490 495
Glu Leu Pro Ala Ala Val Val Val Leu Glu His Gly Lys Thr Met Thr
500 505 510
Glu Lys Glu Ile Val Asp Tyr Val Ala Ser Gln Val Thr Thr Ala Lys
515 520 525
Lys Leu Arg Gly Gly Val Val Phe Val Asp Glu Val Pro Lys Gly Leu
530 535 540
Thr Gly Lys Leu Asp Ala Arg Lys Ile Arg Glu Ile Leu Ile Lys Ala
545 550 555 560
Lys Lys Gly Gly Lys Ile Ala Val
565
<210> 10
<211> 1026
<212> DNA
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> 环子孢子（CS）蛋白（CSP）
<400> 10
atggccaaga agtgtaccat actggtcgtt gcgtcacttc tgttggtcaa ttctctgctc 60
ccaggctatg gacagaacaa atccattcag gcacagagga acctcaacga actctgctac 120
aatgaaggga atgacaataa gctgtatcac gtgctgaatt ccaagaacgg caaaatctac 180
aatcgcaaca cagtaaatcg gttgcttgcc gatgcacccg agggtaagaa gaatgaaaag 240
aagaatgaga aaatcgagcg caacaacaag cttaaacagc caccgcctcc tcctaaccca 300
aatgacccac cgccacccaa tccaaacgac ccaccgcctc ccaaccctaa cgatcctcca 360
ccgcccaacc ctaatgaccc accacctccc aatgcaaacg atccaccccc tcctaacgct 420
aacgaccctg ctccacccaa cgctaacgat cccgcgcccc ccaatgccaa cgaccccgca 480
ccacctaatg ccaacgatcc cgccccgccc aatgctaatg atcctccacc acccaaccca 540
aacgaccctg cccctcctaa tgctaacgat ccaccacctc ccaatccgaa tgatcccgct 600
ccacctcagg ggaacaacaa ccctcagccc caacctagac cacagccgca gccccaaccc 660
caaccccagc cccagcctca accccagccc cagccacgtc cgcagcctca gcctcaacct 720
ggaggcaaca ataacaacaa gaataacaac aacgacgaca gctacattcc aagtgccgag 780
aaaattctgg agtttgttaa gcagatccga gacagcataa ccgaagaatg gtcacagtgt 840
aacgtgacgt gtggatctgg catcagagtg aggaaacgga agggttccaa taagaaagca 900
gaggatctga ctctggagga cattgataca gagatctgca aaatggacaa atgcagctct 960
atcttcaaca tcgtgagtaa tagcctcggg tttgtgattc tgctggtcct ggtgttcttc 1020
aattga 1026
<210> 11
<211> 341
<212> PRT
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> CSP
<400> 11
Met Ala Lys Lys Cys Thr Ile Leu Val Val Ala Ser Leu Leu Leu Val
1 5 10 15
Asn Ser Leu Leu Pro Gly Tyr Gly Gln Asn Lys Ser Ile Gln Ala Gln
20 25 30
Arg Asn Leu Asn Glu Leu Cys Tyr Asn Glu Gly Asn Asp Asn Lys Leu
35 40 45
Tyr His Val Leu Asn Ser Lys Asn Gly Lys Ile Tyr Asn Arg Asn Thr
50 55 60
Val Asn Arg Leu Leu Ala Asp Ala Pro Glu Gly Lys Lys Asn Glu Lys
65 70 75 80
Lys Asn Glu Lys Ile Glu Arg Asn Asn Lys Leu Lys Gln Pro Pro Pro
85 90 95
Pro Pro Asn Pro Asn Asp Pro Pro Pro Pro Asn Pro Asn Asp Pro Pro
100 105 110
Pro Pro Asn Pro Asn Asp Pro Pro Pro Pro Asn Pro Asn Asp Pro Pro
115 120 125
Pro Pro Asn Ala Asn Asp Pro Pro Pro Pro Asn Ala Asn Asp Pro Ala
130 135 140
Pro Pro Asn Ala Asn Asp Pro Ala Pro Pro Asn Ala Asn Asp Pro Ala
145 150 155 160
Pro Pro Asn Ala Asn Asp Pro Ala Pro Pro Asn Ala Asn Asp Pro Pro
165 170 175
Pro Pro Asn Pro Asn Asp Pro Ala Pro Pro Asn Ala Asn Asp Pro Pro
180 185 190
Pro Pro Asn Pro Asn Asp Pro Ala Pro Pro Gln Gly Asn Asn Asn Pro
195 200 205
Gln Pro Gln Pro Arg Pro Gln Pro Gln Pro Gln Pro Gln Pro Gln Pro
210 215 220
Gln Pro Gln Pro Gln Pro Gln Pro Arg Pro Gln Pro Gln Pro Gln Pro
225 230 235 240
Gly Gly Asn Asn Asn Asn Lys Asn Asn Asn Asn Asp Asp Ser Tyr Ile
245 250 255
Pro Ser Ala Glu Lys Ile Leu Glu Phe Val Lys Gln Ile Arg Asp Ser
260 265 270
Ile Thr Glu Glu Trp Ser Gln Cys Asn Val Thr Cys Gly Ser Gly Ile
275 280 285
Arg Val Arg Lys Arg Lys Gly Ser Asn Lys Lys Ala Glu Asp Leu Thr
290 295 300
Leu Glu Asp Ile Asp Thr Glu Ile Cys Lys Met Asp Lys Cys Ser Ser
305 310 315 320
Ile Phe Asn Ile Val Ser Asn Ser Leu Gly Phe Val Ile Leu Leu Val
325 330 335
Leu Val Phe Phe Asn
340
<210> 12
<211> 340
<212> PRT
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> CSP
<400> 12
Met Lys Lys Cys Thr Ile Leu Val Val Ala Ser Leu Leu Leu Val Asn
1 5 10 15
Ser Leu Leu Pro Gly Tyr Gly Gln Asn Lys Ser Ile Gln Ala Gln Arg
20 25 30
Asn Leu Asn Glu Leu Cys Tyr Asn Glu Gly Asn Asp Asn Lys Leu Tyr
35 40 45
His Val Leu Asn Ser Lys Asn Gly Lys Ile Tyr Asn Arg Asn Thr Val
50 55 60
Asn Arg Leu Leu Ala Asp Ala Pro Glu Gly Lys Lys Asn Glu Lys Lys
65 70 75 80
Asn Glu Lys Ile Glu Arg Asn Asn Lys Leu Lys Gln Pro Pro Pro Pro
85 90 95
Pro Asn Pro Asn Asp Pro Pro Pro Pro Asn Pro Asn Asp Pro Pro Pro
100 105 110
Pro Asn Pro Asn Asp Pro Pro Pro Pro Asn Pro Asn Asp Pro Pro Pro
115 120 125
Pro Asn Ala Asn Asp Pro Pro Pro Pro Asn Ala Asn Asp Pro Ala Pro
130 135 140
Pro Asn Ala Asn Asp Pro Ala Pro Pro Asn Ala Asn Asp Pro Ala Pro
145 150 155 160
Pro Asn Ala Asn Asp Pro Ala Pro Pro Asn Ala Asn Asp Pro Pro Pro
165 170 175
Pro Asn Pro Asn Asp Pro Ala Pro Pro Asn Ala Asn Asp Pro Pro Pro
180 185 190
Pro Asn Pro Asn Asp Pro Ala Pro Pro Gln Gly Asn Asn Asn Pro Gln
195 200 205
Pro Gln Pro Arg Pro Gln Pro Gln Pro Gln Pro Gln Pro Gln Pro Gln
210 215 220
Pro Gln Pro Gln Pro Gln Pro Arg Pro Gln Pro Gln Pro Gln Pro Gly
225 230 235 240
Gly Asn Asn Asn Asn Lys Asn Asn Asn Asn Asp Asp Ser Tyr Ile Pro
245 250 255
Ser Ala Glu Lys Ile Leu Glu Phe Val Lys Gln Ile Arg Asp Ser Ile
260 265 270
Thr Glu Glu Trp Ser Gln Cys Asn Val Thr Cys Gly Ser Gly Ile Arg
275 280 285
Val Arg Lys Arg Lys Gly Ser Asn Lys Lys Ala Glu Asp Leu Thr Leu
290 295 300
Glu Asp Ile Asp Thr Glu Ile Cys Lys Met Asp Lys Cys Ser Ser Ile
305 310 315 320
Phe Asn Ile Val Ser Asn Ser Leu Gly Phe Val Ile Leu Leu Val Leu
325 330 335
Val Phe Phe Asn
340
<210> 13
<211> 1203
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> PfCSP人CO
<400> 13
gccaccatgg ctatgagaaa gctcgcgatc ctttccgtga gctcattcct gtttgtcgaa 60
gccttgttcc aggagtatca gtgctatgga tcatcctcca acacaagagt gctcaacgaa 120
ctgaactatg acaatgccgg tactaacctc tacaatgaac tggagatgaa ctactacggc 180
aaacaggaga actggtattc cctgaagaag aattccagat cactgggcga aaacgacgat 240
gggaataatg aggataacga gaagttgcgg aaaccaaagc acaagaagtt gaaacaaccc 300
gccgacggaa accctgatcc taacgccaac ccaaatgtag atcccaacgc caacccaaac 360
gtcgatccca atgccaatcc caatgttgac cccaatgcaa accctaatgc aaatcccaat 420
gccaatccca atgcaaatcc taatgctaat ccaaacgcca accctaacgc gaaccccaac 480
gccaatccta acgcaaatcc taacgcaaat cctaatgcca accctaatgc gaacccgaac 540
gctaatccta acgctaatcc gaatgcaaat ccaaatgcaa atccgaacgc caatcccaac 600
gtagacccaa atgcaaaccc gaacgctaac ccgaacgcaa acccaaacgc caatccgaac 660
gctaatccca atgctaatcc caacgctaac cccaatgcca acccgaatgc caaccccaat 720
gcgaatccaa atgcgaaccc aaacgccaat ccgaatgcga atcctaacgc taacccaaat 780
gctaatccaa acgctaatcc aaatgcgaac cccaatgcga atccaaataa gaacaatcag 840
gggaatggtc agggccataa catgcctaac gaccccaacc gaaatgtgga cgagaacgct 900
aacgcaaaca gcgctgtaaa gaacaacaac aatgaggagc cttctgacaa gcacatcaaa 960
gagtacctga ataaaatcca gaacagtctt tctacggaat ggtccccatg tagtgttact 1020
tgtggcaatg ggattcaagt caggatcaaa ccaggctctg cgaataagcc taaggatgaa 1080
ctggattatg ccaatgacat cgagaagaaa atatgcaaga tggagaagtg cagtagcgtg 1140
ttcaatgtcg taaactcaag cataggtctg ataatggtac tgagctttct gttcctcaac 1200
taa 1203
<210> 14
<211> 398
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> PfCSP
<400> 14
Met Ala Met Arg Lys Leu Ala Ile Leu Ser Val Ser Ser Phe Leu Phe
1 5 10 15
Val Glu Ala Leu Phe Gln Glu Tyr Gln Cys Tyr Gly Ser Ser Ser Asn
20 25 30
Thr Arg Val Leu Asn Glu Leu Asn Tyr Asp Asn Ala Gly Thr Asn Leu
35 40 45
Tyr Asn Glu Leu Glu Met Asn Tyr Tyr Gly Lys Gln Glu Asn Trp Tyr
50 55 60
Ser Leu Lys Lys Asn Ser Arg Ser Leu Gly Glu Asn Asp Asp Gly Asn
65 70 75 80
Asn Glu Asp Asn Glu Lys Leu Arg Lys Pro Lys His Lys Lys Leu Lys
85 90 95
Gln Pro Ala Asp Gly Asn Pro Asp Pro Asn Ala Asn Pro Asn Val Asp
100 105 110
Pro Asn Ala Asn Pro Asn Val Asp Pro Asn Ala Asn Pro Asn Val Asp
115 120 125
Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn
130 135 140
Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn
145 150 155 160
Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn
165 170 175
Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn
180 185 190
Pro Asn Ala Asn Pro Asn Val Asp Pro Asn Ala Asn Pro Asn Ala Asn
195 200 205
Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn
210 215 220
Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn
225 230 235 240
Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn
245 250 255
Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn
260 265 270
Pro Asn Lys Asn Asn Gln Gly Asn Gly Gln Gly His Asn Met Pro Asn
275 280 285
Asp Pro Asn Arg Asn Val Asp Glu Asn Ala Asn Ala Asn Ser Ala Val
290 295 300
Lys Asn Asn Asn Asn Glu Glu Pro Ser Asp Lys His Ile Lys Glu Tyr
305 310 315 320
Leu Asn Lys Ile Gln Asn Ser Leu Ser Thr Glu Trp Ser Pro Cys Ser
325 330 335
Val Thr Cys Gly Asn Gly Ile Gln Val Arg Ile Lys Pro Gly Ser Ala
340 345 350
Asn Lys Pro Lys Asp Glu Leu Asp Tyr Ala Asn Asp Ile Glu Lys Lys
355 360 365
Ile Cys Lys Met Glu Lys Cys Ser Ser Val Phe Asn Val Val Asn Ser
370 375 380
Ser Ile Gly Leu Ile Met Val Leu Ser Phe Leu Phe Leu Asn
385 390 395
<210> 15
<211> 423
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> CSP
<400> 15
Met Arg Lys Leu Ala Ile Leu Ser Val Ser Ser Phe Leu Phe Val Glu
1 5 10 15
Ala Leu Phe Gln Glu Tyr Gln Cys Tyr Gly Ser Ser Ser Asn Thr Arg
20 25 30
Val Leu Asn Glu Leu Asn Tyr Asp Asn Ala Gly Thr Asn Leu Tyr Asn
35 40 45
Glu Leu Glu Met Asn Tyr Tyr Gly Lys Gln Glu Asn Trp Tyr Ser Leu
50 55 60
Lys Lys Asn Ser Arg Ser Leu Gly Glu Asn Asp Asp Gly Asn Asn Asn
65 70 75 80
Asn Gly Asp Asn Gly Arg Glu Gly Lys Asp Glu Asp Lys Arg Asp Gly
85 90 95
Asn Asn Glu Asp Asn Glu Lys Leu Arg Lys Pro Lys His Lys Lys Leu
100 105 110
Lys Gln Pro Gly Asp Gly Asn Pro Asp Pro Asn Ala Asn Pro Asn Val
115 120 125
Asp Pro Asn Ala Asn Pro Asn Val Asp Pro Asn Ala Asn Pro Asn Val
130 135 140
Asp Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala
145 150 155 160
Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala
165 170 175
Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Val
180 185 190
Asp Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala
195 200 205
Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala
210 215 220
Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala
225 230 235 240
Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala
245 250 255
Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala
260 265 270
Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Ala
275 280 285
Asn Pro Asn Ala Asn Pro Asn Ala Asn Pro Asn Lys Asn Asn Gln Gly
290 295 300
Asn Gly Gln Gly His Asn Met Pro Asn Asp Pro Asn Arg Asn Val Asp
305 310 315 320
Glu Asn Ala Asn Ala Asn Asn Ala Val Lys Asn Asn Asn Asn Glu Glu
325 330 335
Pro Ser Asp Lys His Ile Thr Glu Tyr Leu Lys Lys Ile Gln Asn Ser
340 345 350
Leu Ser Thr Glu Trp Ser Pro Cys Ser Val Thr Cys Gly Asn Gly Ile
355 360 365
Gln Val Arg Ile Lys Pro Gly Ser Ala Gly Lys Ser Lys Asp Glu Leu
370 375 380
Asp Tyr Glu Asn Asp Ile Glu Lys Lys Ile Cys Lys Met Glu Lys Cys
385 390 395 400
Ser Ser Val Phe Asn Val Val Asn Ser Ser Ile Gly Leu Ile Met Val
405 410 415
Leu Ser Phe Leu Phe Leu Asn
420
<210> 16
<211> 1143
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> PfCSP人CO
<400> 16
gccaccatgg ctaagaactt cattctgctt gccgtaagct ctattctctt ggtagacttg 60
tttccaacac actgtggaca taatgtagat ctcagtaaag caattaacct caatggtgtt 120
aattttaata atgtagacgc atcctctctc ggcgcggcac acgtgggtca aagtgccagt 180
agggggcgcg gactcggcga gaacccggac gacgaagaag gggatgcgaa aaagaagaag 240
gatgggaaga aggcagaacc caagaatcca agggagaata agctgaagca gccaggagat 300
agagctgacg gccagcctgc cggcgatagg gccgacggac aacccgcggg agaccgcgcg 360
gatggccaac ctgcgggtga ccgcgctgac gggcagcccg caggagacag agccgccgga 420
caacccgccg gcgaccgagc ggatggtcag cccgcgggcg atcgagcgga cggtcagcca 480
gctggagatc gcgcggacgg acaacctgcg ggggacaggg ctgacggtca accagccggt 540
gatagagcag cgggccaacc ggcaggggac cgagccgcag ggcagcccgc cggggacagg 600
gctgacgggc agccagcggg ggatagggcc gccggtcaac ctgcaggcga tcgagctgac 660
ggccaacccg cgggtgaccg ggcagccggc caacctgctg gggatcgagc agacggacag 720
ccggccggag atcgggccgc tggccaaccg gctggagaca gagccgctgg tcaacctgcc 780
ggagatcggg ccgctgggca ggccgcgggt gacagggcag ccggacaagc agcagggggg 840
aacgccggtg gacaaggaca gaacaatgag ggtgcgaatg ccccgaatga aaagagcgtt 900
aaagagtacc tggataaggt tagagcaacc gtaggcaccg aatggacccc ctgctccgtg 960
acttgcgggg taggtgttcg ggtcaggcgg agggtcaatg cggcgaacaa gaaaccggaa 1020
gatctgactt tgaacgatct tgaaacagat gtatgcacga tggacaagtg tgctggcata 1080
tttaacgtag tcagtaactc tctcggcctt gtgatcctct tggtcttggc gctgtttaac 1140
tag 1143
<210> 17
<211> 378
<212> PRT
<213> 间日疟原虫（Plasmodium vivax）
<220>
<221> misc_feature
<223> PvCSP
<400> 17
Met Ala Lys Asn Phe Ile Leu Leu Ala Val Ser Ser Ile Leu Leu Val
1 5 10 15
Asp Leu Phe Pro Thr His Cys Gly His Asn Val Asp Leu Ser Lys Ala
20 25 30
Ile Asn Leu Asn Gly Val Asn Phe Asn Asn Val Asp Ala Ser Ser Leu
35 40 45
Gly Ala Ala His Val Gly Gln Ser Ala Ser Arg Gly Arg Gly Leu Gly
50 55 60
Glu Asn Pro Asp Asp Glu Glu Gly Asp Ala Lys Lys Lys Lys Asp Gly
65 70 75 80
Lys Lys Ala Glu Pro Lys Asn Pro Arg Glu Asn Lys Leu Lys Gln Pro
85 90 95
Gly Asp Arg Ala Asp Gly Gln Pro Ala Gly Asp Arg Ala Asp Gly Gln
100 105 110
Pro Ala Gly Asp Arg Ala Asp Gly Gln Pro Ala Gly Asp Arg Ala Asp
115 120 125
Gly Gln Pro Ala Gly Asp Arg Ala Ala Gly Gln Pro Ala Gly Asp Arg
130 135 140
Ala Asp Gly Gln Pro Ala Gly Asp Arg Ala Asp Gly Gln Pro Ala Gly
145 150 155 160
Asp Arg Ala Asp Gly Gln Pro Ala Gly Asp Arg Ala Asp Gly Gln Pro
165 170 175
Ala Gly Asp Arg Ala Ala Gly Gln Pro Ala Gly Asp Arg Ala Ala Gly
180 185 190
Gln Pro Ala Gly Asp Arg Ala Asp Gly Gln Pro Ala Gly Asp Arg Ala
195 200 205
Ala Gly Gln Pro Ala Gly Asp Arg Ala Asp Gly Gln Pro Ala Gly Asp
210 215 220
Arg Ala Ala Gly Gln Pro Ala Gly Asp Arg Ala Asp Gly Gln Pro Ala
225 230 235 240
Gly Asp Arg Ala Ala Gly Gln Pro Ala Gly Asp Arg Ala Ala Gly Gln
245 250 255
Pro Ala Gly Asp Arg Ala Ala Gly Gln Ala Ala Gly Asp Arg Ala Ala
260 265 270
Gly Gln Ala Ala Gly Gly Asn Ala Gly Gly Gln Gly Gln Asn Asn Glu
275 280 285
Gly Ala Asn Ala Pro Asn Glu Lys Ser Val Lys Glu Tyr Leu Asp Lys
290 295 300
Val Arg Ala Thr Val Gly Thr Glu Trp Thr Pro Cys Ser Val Thr Cys
305 310 315 320
Gly Val Gly Val Arg Val Arg Arg Arg Val Asn Ala Ala Asn Lys Lys
325 330 335
Pro Glu Asp Leu Thr Leu Asn Asp Leu Glu Thr Asp Val Cys Thr Met
340 345 350
Asp Lys Cys Ala Gly Ile Phe Asn Val Val Ser Asn Ser Leu Gly Leu
355 360 365
Val Ile Leu Leu Val Leu Ala Leu Phe Asn
370 375
<210> 18
<211> 368
<212> PRT
<213> 间日疟原虫（Plasmodium vivax）
<220>
<221> misc_feature
<223> CSP
<400> 18
Met Lys Asn Phe Ile Leu Leu Ala Val Ser Ser Ile Leu Leu Val Asp
1 5 10 15
Leu Phe Pro Thr His Cys Gly His Asn Val Asp Leu Ser Lys Ala Ile
20 25 30
Asn Leu Asn Gly Val Asn Phe Asn Asn Val Asp Ala Ser Ser Leu Gly
35 40 45
Ala Ala His Val Gly Gln Ser Ala Ser Arg Gly Arg Gly Leu Gly Glu
50 55 60
Asn Pro Asp Asp Glu Glu Gly Asp Ala Lys Lys Lys Lys Asp Gly Lys
65 70 75 80
Lys Ala Glu Pro Lys Asn Pro Arg Glu Asn Lys Leu Lys Gln Pro Gly
85 90 95
Asp Arg Ala Asp Gly Gln Pro Ala Gly Asp Arg Ala Asp Gly Gln Pro
100 105 110
Ala Gly Asp Arg Ala Asp Gly Gln Pro Ala Gly Asp Arg Ala Gly Gln
115 120 125
Pro Ala Gly Asp Arg Ala Asp Gly Gln Pro Ala Gly Asp Arg Ala Asp
130 135 140
Gly Gln Pro Ala Gly Asp Arg Ala Asp Gly Gln Pro Ala Gly Asp Arg
145 150 155 160
Ala Asp Gly Gln Pro Ala Gly Asp Arg Ala Gly Gln Pro Ala Gly Asp
165 170 175
Arg Ala Gly Gln Pro Ala Gly Asp Arg Ala Asp Gly Gln Pro Ala Gly
180 185 190
Asp Arg Ala Gly Gln Pro Ala Gly Asp Arg Ala Asp Gly Gln Pro Ala
195 200 205
Gly Asp Arg Ala Gly Gln Pro Ala Gly Asp Arg Ala Asp Gly Gln Pro
210 215 220
Ala Gly Asp Arg Ala Gly Gln Pro Ala Gly Asp Arg Ala Gly Gln Pro
225 230 235 240
Ala Gly Asp Arg Ala Gly Gln Pro Ala Gly Asp Arg Ala Gly Gln Pro
245 250 255
Ala Gly Asn Gly Ala Gly Gly Gln Ala Ala Gly Gly Asn Ala Gly Gly
260 265 270
Gln Gly Gln Asn Asn Glu Gly Ala Asn Ala Pro Asn Glu Lys Ser Val
275 280 285
Lys Glu Tyr Leu Asp Lys Val Arg Ala Thr Val Gly Thr Glu Trp Thr
290 295 300
Pro Cys Ser Val Thr Cys Gly Val Gly Val Arg Val Arg Arg Arg Val
305 310 315 320
Asn Ala Ala Asn Lys Lys Pro Glu Asp Leu Thr Leu Asn Asp Leu Glu
325 330 335
Thr Asp Val Cys Thr Met Asp Lys Cys Ala Gly Ile Phe Asn Val Val
340 345 350
Ser Asn Ser Leu Gly Leu Val Ile Leu Leu Val Leu Ala Leu Phe Asn
355 360 365
<210> 19
<211> 1842
<212> DNA
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> 血小板反应蛋白相关匿名蛋白（TRAP）
<400> 19
ggatccgcca ccatggctaa gcttctgggg aacagtaaat acttctttgt ggtcctgctg 60
ctgtgcatta gcgttttcct caacggtcag gagattctgg acgaaatcaa gtactctgag 120
gaggtctgca acgaacaaat cgatctccac attctgctgg atgggagtgg cagcataggt 180
cactctaact ggatcagtca cgtgataccc atgctgacaa cccttgtgga caatttgaac 240
atcagccgcg atgagatcaa tatctccatg accttgttct ccacttatgc cagggaactt 300
gtgagactta agagatatgg gtctacaagc aaagccagtc tgaggttcat catcgcgcaa 360
ctccagaata actattctcc tcatggaacg acaaatctga ctagcgccct gttgaatgtg 420
gacaatctca ttcagaagaa aatgaatcgc cctaatgcca ttcagctcgt gattatcctt 480
actgacggca tccctaacaa tctgaagaag tccactactg ttgtcaacca gctgaagaag 540
aaggacgtca atgtcgctat tattggtgtt ggcgccggag taaacaatat gtttaaccgt 600
atattggtag gatgtggaaa acttgggcct tgtccctact actcttatgg ctcttgggat 660
caagcacaaa ccatgatcaa accatttctc tcaaaggtct gtcaggaagt ggagaaagtg 720
gcactgtgcg gtaagtggga ggagtggagt gagtgttcaa ccacttgcga caacggaacg 780
aaaataagga agcgaaaggt tctccatccc aactgtgccg gggaaatgac agccccatgt 840
aaagtgcggg actgtcctcc caaacctgta gcccctccgg tcattcccat caaagtccct 900
gacgtgcctg tgaaaccagt cgaacctatt gagcccgccg agccagcaga gccagcagaa 960
ccagcagagc ctgcagaacc cgccgaaccc gctgagcccg cggagcccgc cgaacccgct 1020
gaaccggcag aacccgcgga accagcggag cctgcagagc cagctgagcc tgctgaaccg 1080
gcggagcccg ctgaaccagc cgagcctgct aaaccggccg aaccggcaga gcccgctgag 1140
cctgccgagc cagcggaacc agttaacccc gataatccta tcctgccgat caagcccgag 1200
gagccatctg gtggagccga gccattgaat ccagaggtcg agaatccctt tatcatcccc 1260
gacgaaccca tcgaacccat tattgcgcca ggagctgtac cggataagcc catcattcct 1320
gaggaatcaa atgagctgcc aaacaatctt ccagagtctc cctccgatag tcaggtggag 1380
tatcctcggc caaacgacaa tggggataac agcaacaaca caatcaattc caacaagaac 1440
ataccaaata agcatgtgcc tcctacagac gacaacccct acaagggcca ggaagaacga 1500
atccctaagc cgcatcggag caacgacgaa tacatttact acaataatgc taacaataac 1560
gacaagctgg agcccgagat accctctaag gattacgagg aaaacaagag caagaaacag 1620
agcaaaagca acaatggcta taagatcgcc ggcggcataa ttggcgggct ggctattatc 1680
ggctgcattg gagtgggcta taacttcata gccgggtcct ccgccgccgc tatggctgga 1740
gaggcggcac cttttgagga cgtgatggct gatgatgaga aggggatcgt ggaaaacgaa 1800
cagttcaaac tgccagagga caatgattgg aattgactcg ag 1842
<210> 20
<211> 607
<212> PRT
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> TRAP
<400> 20
Met Ala Lys Leu Leu Gly Asn Ser Lys Tyr Phe Phe Val Val Leu Leu
1 5 10 15
Leu Cys Ile Ser Val Phe Leu Asn Gly Gln Glu Ile Leu Asp Glu Ile
20 25 30
Lys Tyr Ser Glu Glu Val Cys Asn Glu Gln Ile Asp Leu His Ile Leu
35 40 45
Leu Asp Gly Ser Gly Ser Ile Gly His Ser Asn Trp Ile Ser His Val
50 55 60
Ile Pro Met Leu Thr Thr Leu Val Asp Asn Leu Asn Ile Ser Arg Asp
65 70 75 80
Glu Ile Asn Ile Ser Met Thr Leu Phe Ser Thr Tyr Ala Arg Glu Leu
85 90 95
Val Arg Leu Lys Arg Tyr Gly Ser Thr Ser Lys Ala Ser Leu Arg Phe
100 105 110
Ile Ile Ala Gln Leu Gln Asn Asn Tyr Ser Pro His Gly Thr Thr Asn
115 120 125
Leu Thr Ser Ala Leu Leu Asn Val Asp Asn Leu Ile Gln Lys Lys Met
130 135 140
Asn Arg Pro Asn Ala Ile Gln Leu Val Ile Ile Leu Thr Asp Gly Ile
145 150 155 160
Pro Asn Asn Leu Lys Lys Ser Thr Thr Val Val Asn Gln Leu Lys Lys
165 170 175
Lys Asp Val Asn Val Ala Ile Ile Gly Val Gly Ala Gly Val Asn Asn
180 185 190
Met Phe Asn Arg Ile Leu Val Gly Cys Gly Lys Leu Gly Pro Cys Pro
195 200 205
Tyr Tyr Ser Tyr Gly Ser Trp Asp Gln Ala Gln Thr Met Ile Lys Pro
210 215 220
Phe Leu Ser Lys Val Cys Gln Glu Val Glu Lys Val Ala Leu Cys Gly
225 230 235 240
Lys Trp Glu Glu Trp Ser Glu Cys Ser Thr Thr Cys Asp Asn Gly Thr
245 250 255
Lys Ile Arg Lys Arg Lys Val Leu His Pro Asn Cys Ala Gly Glu Met
260 265 270
Thr Ala Pro Cys Lys Val Arg Asp Cys Pro Pro Lys Pro Val Ala Pro
275 280 285
Pro Val Ile Pro Ile Lys Val Pro Asp Val Pro Val Lys Pro Val Glu
290 295 300
Pro Ile Glu Pro Ala Glu Pro Ala Glu Pro Ala Glu Pro Ala Glu Pro
305 310 315 320
Ala Glu Pro Ala Glu Pro Ala Glu Pro Ala Glu Pro Ala Glu Pro Ala
325 330 335
Glu Pro Ala Glu Pro Ala Glu Pro Ala Glu Pro Ala Glu Pro Ala Glu
340 345 350
Pro Ala Glu Pro Ala Glu Pro Ala Glu Pro Ala Glu Pro Ala Lys Pro
355 360 365
Ala Glu Pro Ala Glu Pro Ala Glu Pro Ala Glu Pro Ala Glu Pro Val
370 375 380
Asn Pro Asp Asn Pro Ile Leu Pro Ile Lys Pro Glu Glu Pro Ser Gly
385 390 395 400
Gly Ala Glu Pro Leu Asn Pro Glu Val Glu Asn Pro Phe Ile Ile Pro
405 410 415
Asp Glu Pro Ile Glu Pro Ile Ile Ala Pro Gly Ala Val Pro Asp Lys
420 425 430
Pro Ile Ile Pro Glu Glu Ser Asn Glu Leu Pro Asn Asn Leu Pro Glu
435 440 445
Ser Pro Ser Asp Ser Gln Val Glu Tyr Pro Arg Pro Asn Asp Asn Gly
450 455 460
Asp Asn Ser Asn Asn Thr Ile Asn Ser Asn Lys Asn Ile Pro Asn Lys
465 470 475 480
His Val Pro Pro Thr Asp Asp Asn Pro Tyr Lys Gly Gln Glu Glu Arg
485 490 495
Ile Pro Lys Pro His Arg Ser Asn Asp Glu Tyr Ile Tyr Tyr Asn Asn
500 505 510
Ala Asn Asn Asn Asp Lys Leu Glu Pro Glu Ile Pro Ser Lys Asp Tyr
515 520 525
Glu Glu Asn Lys Ser Lys Lys Gln Ser Lys Ser Asn Asn Gly Tyr Lys
530 535 540
Ile Ala Gly Gly Ile Ile Gly Gly Leu Ala Ile Ile Gly Cys Ile Gly
545 550 555 560
Val Gly Tyr Asn Phe Ile Ala Gly Ser Ser Ala Ala Ala Met Ala Gly
565 570 575
Glu Ala Ala Pro Phe Glu Asp Val Met Ala Asp Asp Glu Lys Gly Ile
580 585 590
Val Glu Asn Glu Gln Phe Lys Leu Pro Glu Asp Asn Asp Trp Asn
595 600 605
<210> 21
<211> 606
<212> PRT
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> TRAP
<400> 21
Met Lys Leu Leu Gly Asn Ser Lys Tyr Phe Phe Val Val Leu Leu Leu
1 5 10 15
Cys Ile Ser Val Phe Leu Asn Gly Gln Glu Ile Leu Asp Glu Ile Lys
20 25 30
Tyr Ser Glu Glu Val Cys Asn Glu Gln Ile Asp Leu His Ile Leu Leu
35 40 45
Asp Gly Ser Gly Ser Ile Gly His Ser Asn Trp Ile Ser His Val Ile
50 55 60
Pro Met Leu Thr Thr Leu Val Asp Asn Leu Asn Ile Ser Arg Asp Glu
65 70 75 80
Ile Asn Ile Ser Met Thr Leu Phe Ser Thr Tyr Ala Arg Glu Leu Val
85 90 95
Arg Leu Lys Arg Tyr Gly Ser Thr Ser Lys Ala Ser Leu Arg Phe Ile
100 105 110
Ile Ala Gln Leu Gln Asn Asn Tyr Ser Pro His Gly Thr Thr Asn Leu
115 120 125
Thr Ser Ala Leu Leu Asn Val Asp Asn Leu Ile Gln Lys Lys Met Asn
130 135 140
Arg Pro Asn Ala Ile Gln Leu Val Ile Ile Leu Thr Asp Gly Ile Pro
145 150 155 160
Asn Asn Leu Lys Lys Ser Thr Thr Val Val Asn Gln Leu Lys Lys Lys
165 170 175
Asp Val Asn Val Ala Ile Ile Gly Val Gly Ala Gly Val Asn Asn Met
180 185 190
Phe Asn Arg Ile Leu Val Gly Cys Gly Lys Leu Gly Pro Cys Pro Tyr
195 200 205
Tyr Ser Tyr Gly Ser Trp Asp Gln Ala Gln Thr Met Ile Lys Pro Phe
210 215 220
Leu Ser Lys Val Cys Gln Glu Val Glu Lys Val Ala Leu Cys Gly Lys
225 230 235 240
Trp Glu Glu Trp Ser Glu Cys Ser Thr Thr Cys Asp Asn Gly Thr Lys
245 250 255
Ile Arg Lys Arg Lys Val Leu His Pro Asn Cys Ala Gly Glu Met Thr
260 265 270
Ala Pro Cys Lys Val Arg Asp Cys Pro Pro Lys Pro Val Ala Pro Pro
275 280 285
Val Ile Pro Ile Lys Val Pro Asp Val Pro Val Lys Pro Val Glu Pro
290 295 300
Ile Glu Pro Ala Glu Pro Ala Glu Pro Ala Glu Pro Ala Glu Pro Ala
305 310 315 320
Glu Pro Ala Glu Pro Ala Glu Pro Ala Glu Pro Ala Glu Pro Ala Glu
325 330 335
Pro Ala Glu Pro Ala Glu Pro Ala Glu Pro Ala Glu Pro Ala Glu Pro
340 345 350
Ala Glu Pro Ala Glu Pro Ala Glu Pro Ala Glu Pro Ala Lys Pro Ala
355 360 365
Glu Pro Ala Glu Pro Ala Glu Pro Ala Glu Pro Ala Glu Pro Val Asn
370 375 380
Pro Asp Asn Pro Ile Leu Pro Ile Lys Pro Glu Glu Pro Ser Gly Gly
385 390 395 400
Ala Glu Pro Leu Asn Pro Glu Val Glu Asn Pro Phe Ile Ile Pro Asp
405 410 415
Glu Pro Ile Glu Pro Ile Ile Ala Pro Gly Ala Val Pro Asp Lys Pro
420 425 430
Ile Ile Pro Glu Glu Ser Asn Glu Leu Pro Asn Asn Leu Pro Glu Ser
435 440 445
Pro Ser Asp Ser Gln Val Glu Tyr Pro Arg Pro Asn Asp Asn Gly Asp
450 455 460
Asn Ser Asn Asn Thr Ile Asn Ser Asn Lys Asn Ile Pro Asn Lys His
465 470 475 480
Val Pro Pro Thr Asp Asp Asn Pro Tyr Lys Gly Gln Glu Glu Arg Ile
485 490 495
Pro Lys Pro His Arg Ser Asn Asp Glu Tyr Ile Tyr Tyr Asn Asn Ala
500 505 510
Asn Asn Asn Asp Lys Leu Glu Pro Glu Ile Pro Ser Lys Asp Tyr Glu
515 520 525
Glu Asn Lys Ser Lys Lys Gln Ser Lys Ser Asn Asn Gly Tyr Lys Ile
530 535 540
Ala Gly Gly Ile Ile Gly Gly Leu Ala Ile Ile Gly Cys Ile Gly Val
545 550 555 560
Gly Tyr Asn Phe Ile Ala Gly Ser Ser Ala Ala Ala Met Ala Gly Glu
565 570 575
Ala Ala Pro Phe Glu Asp Val Met Ala Asp Asp Glu Lys Gly Ile Val
580 585 590
Glu Asn Glu Gln Phe Lys Leu Pro Glu Asp Asn Asp Trp Asn
595 600 605
<210> 22
<211> 1734
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 血小板反应蛋白相关匿名蛋白（TRAP）
<400> 22
gccaccatgg ctaatcactt gggcaacgtg aagtacctgg tgatagtgtt cctgatcttc 60
ttcgacctgt ttctggtcaa cggcagagat gtgcagaaca acatagtcga cgagattaag 120
tacagagagg aggtgtgcaa tgacgaggtg gatctctacc tgctgatgga ctgtagcggg 180
tcaatacgac ggcacaattg ggttaatcat gctgtcccct tggctatgaa gctgatccag 240
cagctcaatc tgaacgataa tgccattcac ctgtatgctt ccgtgttcag caataatgca 300
agggagatca ttcgcctcca cagtgacgct agtaagaata aggagaaagc cctgatcatc 360
atcaaatccc tgcttagcac caaccttccc tatggcaaga ccaatctcac agacgcgctt 420
ctgcaggtaa ggaagcatct gaacgatcgc atcaacagag agaatgcgaa tcagctggta 480
gtaatcctga ctgatgggat tcccgattcc atccaggaca gcctgaagga atcaaggaag 540
ctcagcgata gaggggtgaa aatcgcagtt ttcggaatcg gtcaggggat taacgtggcc 600
tttaatcgct ttctggtggg atgtcatcca tctgatggga aatgcaatct gtatgccgat 660
tctgcgtggg agaatgtgaa gaacgtgatt ggccccttta tgaaagccgt gtgcgtagag 720
gtggagaaaa ccgcctcctg tggagtttgg gacgaatgga gcccctgttc cgttacttgc 780
gggaaaggga cccgaagtcg caagagagaa atccttcacg agggttgcac ttctgaactc 840
caagagcaat gcgaggagga aaggtgtctc cctaaacgag aaccccttga tgtgcctgac 900
gaacctgagg acgatcaacc aagaccaagg ggagacaact tcgccgtcga gaaacccaat 960
gagaatatca ttgacaacaa tccacaggag ccatcaccta atccagagga aggcaaaggc 1020
gaaaacccta acggttttga cttggacgag aatccggaaa atccacccaa cccgcctaac 1080
cctccaaatc caccaaaccc tccgaatcct cctaaccctg atatccccga gcaagagcct 1140
aatattccag aggatagcga aaaggaggtc ccctctgacg tccctaagaa ccctgaagat 1200
gatcgcgaag agaactttga catacccaag aaacccgaga ataaacacga taatcagaac 1260
aatctcccga acgataaatc tgaccgctac attccataca gtccattgag cccaaaggtc 1320
ttggataatg aacggaaaca gtccgaccct caatcacagg acaataacgg taatcgtcat 1380
gttccgaact ccgaagatcg ggaaacaagg ccccatgggc gtaacaacga aaaccggagc 1440
tataaccgga agcacaacaa cacacccaag cacccggaac gggaagaaca tgagaagccc 1500
gataacaaca agaagaaggc aggctctgac aacaaataca aaatagccgg aggtattgcc 1560
ggaggattgg cactgctggc ctgcgctggc cttgcttaca agttcgtcgt tccaggcgcc 1620
gcaacgccct atgcaggcga accagctccc tttgacgaga cactgggaga agaggacaaa 1680
gacctggacg agcctgaaca gttcagactg cccgaggaaa acgagtggaa ttaa 1734
<210> 23
<211> 575
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> PfTRAP
<400> 23
Met Ala Asn His Leu Gly Asn Val Lys Tyr Leu Val Ile Val Phe Leu
1 5 10 15
Ile Phe Phe Asp Leu Phe Leu Val Asn Gly Arg Asp Val Gln Asn Asn
20 25 30
Ile Val Asp Glu Ile Lys Tyr Arg Glu Glu Val Cys Asn Asp Glu Val
35 40 45
Asp Leu Tyr Leu Leu Met Asp Cys Ser Gly Ser Ile Arg Arg His Asn
50 55 60
Trp Val Asn His Ala Val Pro Leu Ala Met Lys Leu Ile Gln Gln Leu
65 70 75 80
Asn Leu Asn Asp Asn Ala Ile His Leu Tyr Ala Ser Val Phe Ser Asn
85 90 95
Asn Ala Arg Glu Ile Ile Arg Leu His Ser Asp Ala Ser Lys Asn Lys
100 105 110
Glu Lys Ala Leu Ile Ile Ile Lys Ser Leu Leu Ser Thr Asn Leu Pro
115 120 125
Tyr Gly Lys Thr Asn Leu Thr Asp Ala Leu Leu Gln Val Arg Lys His
130 135 140
Leu Asn Asp Arg Ile Asn Arg Glu Asn Ala Asn Gln Leu Val Val Ile
145 150 155 160
Leu Thr Asp Gly Ile Pro Asp Ser Ile Gln Asp Ser Leu Lys Glu Ser
165 170 175
Arg Lys Leu Ser Asp Arg Gly Val Lys Ile Ala Val Phe Gly Ile Gly
180 185 190
Gln Gly Ile Asn Val Ala Phe Asn Arg Phe Leu Val Gly Cys His Pro
195 200 205
Ser Asp Gly Lys Cys Asn Leu Tyr Ala Asp Ser Ala Trp Glu Asn Val
210 215 220
Lys Asn Val Ile Gly Pro Phe Met Lys Ala Val Cys Val Glu Val Glu
225 230 235 240
Lys Thr Ala Ser Cys Gly Val Trp Asp Glu Trp Ser Pro Cys Ser Val
245 250 255
Thr Cys Gly Lys Gly Thr Arg Ser Arg Lys Arg Glu Ile Leu His Glu
260 265 270
Gly Cys Thr Ser Glu Leu Gln Glu Gln Cys Glu Glu Glu Arg Cys Leu
275 280 285
Pro Lys Arg Glu Pro Leu Asp Val Pro Asp Glu Pro Glu Asp Asp Gln
290 295 300
Pro Arg Pro Arg Gly Asp Asn Phe Ala Val Glu Lys Pro Asn Glu Asn
305 310 315 320
Ile Ile Asp Asn Asn Pro Gln Glu Pro Ser Pro Asn Pro Glu Glu Gly
325 330 335
Lys Gly Glu Asn Pro Asn Gly Phe Asp Leu Asp Glu Asn Pro Glu Asn
340 345 350
Pro Pro Asn Pro Pro Asn Pro Pro Asn Pro Pro Asn Pro Pro Asn Pro
355 360 365
Pro Asn Pro Asp Ile Pro Glu Gln Glu Pro Asn Ile Pro Glu Asp Ser
370 375 380
Glu Lys Glu Val Pro Ser Asp Val Pro Lys Asn Pro Glu Asp Asp Arg
385 390 395 400
Glu Glu Asn Phe Asp Ile Pro Lys Lys Pro Glu Asn Lys His Asp Asn
405 410 415
Gln Asn Asn Leu Pro Asn Asp Lys Ser Asp Arg Tyr Ile Pro Tyr Ser
420 425 430
Pro Leu Ser Pro Lys Val Leu Asp Asn Glu Arg Lys Gln Ser Asp Pro
435 440 445
Gln Ser Gln Asp Asn Asn Gly Asn Arg His Val Pro Asn Ser Glu Asp
450 455 460
Arg Glu Thr Arg Pro His Gly Arg Asn Asn Glu Asn Arg Ser Tyr Asn
465 470 475 480
Arg Lys His Asn Asn Thr Pro Lys His Pro Glu Arg Glu Glu His Glu
485 490 495
Lys Pro Asp Asn Asn Lys Lys Lys Ala Gly Ser Asp Asn Lys Tyr Lys
500 505 510
Ile Ala Gly Gly Ile Ala Gly Gly Leu Ala Leu Leu Ala Cys Ala Gly
515 520 525
Leu Ala Tyr Lys Phe Val Val Pro Gly Ala Ala Thr Pro Tyr Ala Gly
530 535 540
Glu Pro Ala Pro Phe Asp Glu Thr Leu Gly Glu Glu Asp Lys Asp Leu
545 550 555 560
Asp Glu Pro Glu Gln Phe Arg Leu Pro Glu Glu Asn Glu Trp Asn
565 570 575
<210> 24
<211> 560
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> TRAP
<400> 24
Met Asn His Leu Gly Asn Val Lys Tyr Leu Val Ile Val Phe Leu Ile
1 5 10 15
Phe Phe Asp Leu Phe Leu Val Asn Gly Arg Asp Val Gln Asn Asn Ile
20 25 30
Val Asp Glu Ile Lys Tyr Arg Glu Glu Val Cys Asn Asp Glu Val Asp
35 40 45
Leu Tyr Leu Leu Met Asp Cys Ser Gly Ser Ile Arg Arg His Asn Trp
50 55 60
Val Asn His Ala Val Pro Leu Ala Met Lys Leu Ile Gln Gln Leu Asn
65 70 75 80
Leu Asn Glu Asn Ala Ile His Leu Tyr Ala Asn Val Phe Ser Asn Asn
85 90 95
Ala Arg Glu Ile Ile Arg Leu His Ser Asp Ala Ser Lys Asn Lys Glu
100 105 110
Lys Ala Leu Ser Ile Ile Lys Ser Leu Leu Ser Thr Asn Leu Pro Phe
115 120 125
Gly Arg Thr Asn Leu Thr Asp Ala Leu Leu Gln Val Arg Lys His Leu
130 135 140
Asn Asp Arg Ile Asn Arg Glu Asn Ala Asn Gln Leu Val Val Ile Leu
145 150 155 160
Thr Asp Gly Ile Pro Asn Ser Ile Gln Asp Ser Leu Lys Glu Ser Arg
165 170 175
Lys Leu Ser Asp Arg Gly Val Lys Ile Ala Val Phe Gly Ile Gly Gln
180 185 190
Gly Ile Asn Val Ala Phe Asn Arg Phe Leu Val Gly Cys His Pro Ser
195 200 205
Asp Gly Lys Cys Asn Leu Tyr Ala Asp Ser Ala Trp Glu Asn Val Lys
210 215 220
Asn Val Ile Gly Pro Phe Met Lys Ala Val Cys Val Glu Val Glu Lys
225 230 235 240
Thr Ala Ser Cys Gly Val Trp Asp Glu Trp Ser Pro Cys Ser Val Thr
245 250 255
Cys Gly Lys Gly Thr Arg Ser Arg Lys Arg Glu Ile Leu His Glu Gly
260 265 270
Cys Thr Ser Glu Leu Gln Glu Gln Cys Glu Glu Glu Arg Cys Pro Pro
275 280 285
Lys Trp Glu Pro Leu Asp Val Pro Asp Asp Pro Glu Asp Asp Gln Pro
290 295 300
Arg Pro Arg Gly Asp Asn Ser Ser Val Gln Lys Pro Glu Glu Asn Ile
305 310 315 320
Ile Asp Asn Asn Pro Gln Glu Pro Ser Pro Asn Pro Glu Glu Gly Lys
325 330 335
Gly Glu Asn Pro Asn Gly Phe Asp Leu Asp Glu Asn Pro Glu Asn Pro
340 345 350
Pro Asn Pro Asp Ile Pro Gln Gln Glu Pro Asn Ile Pro Glu Asp Ser
355 360 365
Glu Lys Glu Val Pro Ser Asp Val Pro Lys Asn Pro Glu Asp Asp Arg
370 375 380
Glu Glu Asn Phe Asp Ile Pro Lys Lys Pro Glu Asn Lys His Asp Asn
385 390 395 400
Gln Asn Asn Leu Pro Asn Asp Lys Ser Asp Arg Tyr Ile Pro Tyr Ser
405 410 415
Pro Leu Pro Pro Asn Val Leu Asp Asn Glu Arg Lys Gln Ser Asp Pro
420 425 430
Gln Ser Gln Asp Asn Asn Gly Asn Arg His Val Pro Asn Ser Glu Asp
435 440 445
Arg Glu Thr Arg Pro His Gly Arg Asn Asn Glu Asn Arg Ser Tyr Asn
450 455 460
Arg Lys His Asn Asp Thr Pro Lys His Pro Glu Arg Glu Glu His Glu
465 470 475 480
Lys Pro Asp Asn Asn Lys Lys Lys Gly Gly Ser Ser Asp Asn Lys Tyr
485 490 495
Lys Ile Ala Gly Gly Ile Ala Gly Gly Leu Ala Leu Leu Ala Cys Ala
500 505 510
Gly Leu Ala Tyr Lys Phe Val Val Pro Gly Ala Ala Thr Pro Tyr Ala
515 520 525
Gly Glu Pro Ala Pro Phe Asp Glu Thr Leu Gly Glu Glu Asp Lys Asp
530 535 540
Leu Asp Glu Pro Glu Gln Phe Arg Leu Pro Glu Glu Asn Glu Trp Asn
545 550 555 560
<210> 25
<211> 1680
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> PvTRAP人CO
<400> 25
gccaccatgg ctaagctgct gcagaacaag tcttatctcc tggtcgtgtt cctgctttac 60
gtatccatat ttgcacgggg agatgaaaaa gttgtagatg aggtaaagta cagcgaggag 120
gtatgcaacg aatctgtcga tctctatctt ctcgtagatg gctctggtag tattgggtat 180
ccgaactgga taactaaggt aattcctatg ctgaacggtc ttattaatag tttgagcctc 240
agtcgagaca cgatcaatct ctatatgaat ctcttcggaa attacactac agaactcata 300
cggctgggct ctgggcagag tatagataaa cgacaggctc tgtcaaaggt gactgaactt 360
agaaagacct acacgccata tggcactact aacatgacag ccgcgcttga tgaagtccaa 420
aagcacctta atgatagagt gaaccgagag aaagcgatac aacttgtcat attgatgacc 480
gacggggtgc ccaactcaaa atacagggcg ctggaggtcg caaataaact gaagcaacgc 540
aatgtttcct tggccgttat tggtgtcgga caaggaatca accatcagtt caatcggctg 600
atcgcgggct gccgacctcg ggagccgaac tgcaaatttt actcttatgc tgattggaat 660
gaggccgtag cacttataaa gccctttatt gctaaggtat gcacagaagt tgagcgcgtt 720
gccaactgtg gaccctggga cccctggacc gcgtgtagcg ttacgtgcgg caggggaacg 780
cactccaggt ctcgcccttc attgcatgaa aagtgcacta cccatatggt gtctgaatgt 840
gaagaggggg aatgcccagt ggagcctgag ccactgccag tacctgcgcc actcccaacg 900
gttccagaag atgtaaatcc gcgagacacg gacgacgaaa atgagaaccc gaacttcaac 960
aagggactgg acgtgccgga tgaagatgat gacgaagtac cgcccgcgaa tgaaggtgcg 1020
gatggcaacc cagtcgagga aaatgttttc ccgcctgctg acgatagcgt tcctgatgag 1080
agtaacgtgc tgcctctccc tccggcagtc ccaggcgggt cctctgagga gttcccggcg 1140
gacgttcaga acaatcccga ctctcctgaa gagctgccga tggagcaaga agtgcctcaa 1200
gataataacg tgaatgaacc agagagatcc gatagcaacg gttatggcgt aaatgagaag 1260
gtgataccta atcccctcga caatgagcga gacatggcca ataagaataa gaccgttcac 1320
ccaggccgga aggacagcgc gagagatcga tatgcccgcc cccatggttc cactcatgtc 1380
aataataata gagccaatga gaactcagac ataccaaaca acccagtacc ttctgattac 1440
gaacagccgg aggacaaggc taaaaagtcc tccaataacg gctataaaat cgctggtgga 1500
gtgatcgctg ggctcgcgtt ggttgggtgc gtcggtttcg cgtataactt cgtagcgggt 1560
ggaggcgctg ctggtatggc aggcgaaccg gctccctttg acgaagctat ggccgaagac 1620
gaaaaagacg tagccgaagc ggaccagttc aaattgcctg aggacaacga ttggaattaa 1680
<210> 26
<211> 557
<212> PRT
<213> 间日疟原虫（Plasmodium vivax）
<220>
<221> misc_feature
<223> PvTRAP
<400> 26
Met Ala Lys Leu Leu Gln Asn Lys Ser Tyr Leu Leu Val Val Phe Leu
1 5 10 15
Leu Tyr Val Ser Ile Phe Ala Arg Gly Asp Glu Lys Val Val Asp Glu
20 25 30
Val Lys Tyr Ser Glu Glu Val Cys Asn Glu Ser Val Asp Leu Tyr Leu
35 40 45
Leu Val Asp Gly Ser Gly Ser Ile Gly Tyr Pro Asn Trp Ile Thr Lys
50 55 60
Val Ile Pro Met Leu Asn Gly Leu Ile Asn Ser Leu Ser Leu Ser Arg
65 70 75 80
Asp Thr Ile Asn Leu Tyr Met Asn Leu Phe Gly Asn Tyr Thr Thr Glu
85 90 95
Leu Ile Arg Leu Gly Ser Gly Gln Ser Ile Asp Lys Arg Gln Ala Leu
100 105 110
Ser Lys Val Thr Glu Leu Arg Lys Thr Tyr Thr Pro Tyr Gly Thr Thr
115 120 125
Asn Met Thr Ala Ala Leu Asp Glu Val Gln Lys His Leu Asn Asp Arg
130 135 140
Val Asn Arg Glu Lys Ala Ile Gln Leu Val Ile Leu Met Thr Asp Gly
145 150 155 160
Val Pro Asn Ser Lys Tyr Arg Ala Leu Glu Val Ala Asn Lys Leu Lys
165 170 175
Gln Arg Asn Val Ser Leu Ala Val Ile Gly Val Gly Gln Gly Ile Asn
180 185 190
His Gln Phe Asn Arg Leu Ile Ala Gly Cys Arg Pro Arg Glu Pro Asn
195 200 205
Cys Lys Phe Tyr Ser Tyr Ala Asp Trp Asn Glu Ala Val Ala Leu Ile
210 215 220
Lys Pro Phe Ile Ala Lys Val Cys Thr Glu Val Glu Arg Val Ala Asn
225 230 235 240
Cys Gly Pro Trp Asp Pro Trp Thr Ala Cys Ser Val Thr Cys Gly Arg
245 250 255
Gly Thr His Ser Arg Ser Arg Pro Ser Leu His Glu Lys Cys Thr Thr
260 265 270
His Met Val Ser Glu Cys Glu Glu Gly Glu Cys Pro Val Glu Pro Glu
275 280 285
Pro Leu Pro Val Pro Ala Pro Leu Pro Thr Val Pro Glu Asp Val Asn
290 295 300
Pro Arg Asp Thr Asp Asp Glu Asn Glu Asn Pro Asn Phe Asn Lys Gly
305 310 315 320
Leu Asp Val Pro Asp Glu Asp Asp Asp Glu Val Pro Pro Ala Asn Glu
325 330 335
Gly Ala Asp Gly Asn Pro Val Glu Glu Asn Val Phe Pro Pro Ala Asp
340 345 350
Asp Ser Val Pro Asp Glu Ser Asn Val Leu Pro Leu Pro Pro Ala Val
355 360 365
Pro Gly Gly Ser Ser Glu Glu Phe Pro Ala Asp Val Gln Asn Asn Pro
370 375 380
Asp Ser Pro Glu Glu Leu Pro Met Glu Gln Glu Val Pro Gln Asp Asn
385 390 395 400
Asn Val Asn Glu Pro Glu Arg Ser Asp Ser Asn Gly Tyr Gly Val Asn
405 410 415
Glu Lys Val Ile Pro Asn Pro Leu Asp Asn Glu Arg Asp Met Ala Asn
420 425 430
Lys Asn Lys Thr Val His Pro Gly Arg Lys Asp Ser Ala Arg Asp Arg
435 440 445
Tyr Ala Arg Pro His Gly Ser Thr His Val Asn Asn Asn Arg Ala Asn
450 455 460
Glu Asn Ser Asp Ile Pro Asn Asn Pro Val Pro Ser Asp Tyr Glu Gln
465 470 475 480
Pro Glu Asp Lys Ala Lys Lys Ser Ser Asn Asn Gly Tyr Lys Ile Ala
485 490 495
Gly Gly Val Ile Ala Gly Leu Ala Leu Val Gly Cys Val Gly Phe Ala
500 505 510
Tyr Asn Phe Val Ala Gly Gly Gly Ala Ala Gly Met Ala Gly Glu Pro
515 520 525
Ala Pro Phe Asp Glu Ala Met Ala Glu Asp Glu Lys Asp Val Ala Glu
530 535 540
Ala Asp Gln Phe Lys Leu Pro Glu Asp Asn Asp Trp Asn
545 550 555
<210> 27
<211> 556
<212> PRT
<213> 间日疟原虫（Plasmodium vivax）
<220>
<221> misc_feature
<223> TRAP
<400> 27
Met Lys Leu Leu Gln Asn Lys Ser Tyr Leu Leu Val Val Phe Leu Leu
1 5 10 15
Tyr Val Ser Ile Phe Ala Arg Gly Asp Glu Lys Val Val Asp Glu Val
20 25 30
Lys Tyr Ser Glu Glu Val Cys Asn Glu Ser Val Asp Leu Tyr Leu Leu
35 40 45
Val Asp Gly Ser Gly Ser Ile Gly Tyr Pro Asn Trp Ile Thr Lys Val
50 55 60
Ile Pro Met Leu Asn Gly Leu Ile Asn Ser Leu Ser Leu Ser Arg Asp
65 70 75 80
Thr Ile Asn Leu Tyr Met Asn Leu Phe Gly Asn Tyr Thr Thr Glu Leu
85 90 95
Ile Arg Leu Gly Ser Gly Gln Ser Ile Asp Lys Arg Gln Ala Leu Ser
100 105 110
Lys Val Thr Glu Leu Arg Lys Ser Tyr Ser Pro Tyr Gly Thr Thr Asn
115 120 125
Met Thr Ala Ala Leu Asp Glu Val Gln Lys His Leu Asn Asp Arg Val
130 135 140
Asn Arg Glu Lys Ala Ile Gln Leu Val Ile Leu Met Thr Asp Gly Ile
145 150 155 160
Pro Asn Ser Lys Tyr Thr Ala Leu Glu Val Ala Lys Lys Leu Lys Gln
165 170 175
Arg Asn Val Ser Leu Ala Val Ile Gly Ile Gly Gln Gly Ile Asn His
180 185 190
Gln Phe Asn Arg Leu Ile Ala Gly Cys Arg Pro Arg Glu Ser Asn Cys
195 200 205
Lys Phe Tyr Ser Tyr Ala Asp Trp Asn Glu Ala Val Ala Leu Ile Lys
210 215 220
Pro Phe Ile Ala Lys Val Cys Thr Glu Val Glu Arg Val Ala Asn Cys
225 230 235 240
Gly Pro Trp Asp Pro Trp Thr Ala Cys Ser Val Thr Cys Gly Arg Gly
245 250 255
Thr His Ser Arg Ser Arg Pro Ser Leu His Glu Gly Cys Thr Thr His
260 265 270
Met Val Ser Glu Cys Glu Glu Gly Glu Cys Pro Val Glu Pro Glu Pro
275 280 285
Leu Pro Val Pro Ala Pro Leu Pro Thr Val Pro Glu Asp Val Asn Pro
290 295 300
Arg Asp Thr Asp Asp Glu Asn Glu Asn Pro Asn Phe Asn Lys Gly Leu
305 310 315 320
Asp Val Pro Glu Glu Asp Asp Asp Glu Val Pro Pro Ala Asn Glu Arg
325 330 335
Ala Asp Gly Asn Pro Val Glu Glu Asn Val Phe Pro Pro Ala Asp Asp
340 345 350
Ser Val Pro Asp Glu Ser Asn Val Leu Pro Leu Pro Pro Ala Val Pro
355 360 365
Gly Gly Ser Ser Glu Glu Phe Pro Ala Asp Val Gln Asn Asn Pro Asp
370 375 380
Ser Pro Glu Glu Leu Pro Met Glu Gln Glu Val Pro Gln Asp Asn Asn
385 390 395 400
Val Asn Glu Pro Glu Arg Ser Asp Ser Asn Gly Tyr Gly Val Asn Glu
405 410 415
Lys Val Ile Pro Asn Pro Leu Asp Asn Glu Arg Asp Met Ala Asn Lys
420 425 430
Asn Lys Thr Val His Pro Asp Arg Lys Asp Ser Ala Arg Asp Arg Tyr
435 440 445
Ala Arg Pro His Gly Ser Thr His Val Asn Asn Asn Arg Ala Asn Glu
450 455 460
Asn Ser Asp Ile Pro Asn Asn Pro Val Pro Ser Asp Tyr Glu Gln Pro
465 470 475 480
Glu Asp Lys Ala Lys Lys Ser Ser Asn Asn Gly Tyr Lys Ile Ala Gly
485 490 495
Gly Val Ile Ala Gly Leu Ala Leu Val Gly Cys Val Gly Phe Ala Tyr
500 505 510
Asn Phe Val Ala Gly Gly Gly Ala Ala Gly Met Ala Gly Glu Pro Ala
515 520 525
Pro Phe Asp Glu Ala Met Ala Glu Asp Glu Lys Asp Val Ala Glu Ala
530 535 540
Asp Gln Phe Lys Leu Pro Glu Asp Asn Asp Trp Asn
545 550 555
<210> 28
<211> 1086
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 半胱氨酸蛋白酶抑制剂（ICP）
<400> 28
ggatccgcca ccatggctaa atccattacc ttctttgtgt tcaacatttg cagcattttg 60
gctctgctga gtcactgtga ggacaatgac atctacagct tcgacattgt caatgagaca 120
aattggctga agatcgccaa gaacatcttc aaaggcaagt ctcctagcaa tttcacgatc 180
ataccgttta acaataccgg tagttctaac gataacgagt caaacaagga ggaatcagta 240
ctgctgatca gaaagaagat caaaagcaac aagaatcacg atagttccat cattagtggt 300
gacactgtta acggggacat tagtgacctg aattatacgg ctagcaactt ttccgataac 360
tctgaggaca tagaagataa ccagaaatat cccacaacca gctacaatag tttcaaccat 420
ctcaattcca atatcgcctt taacgaagag tccgaataca ttgagattaa tagcgagtct 480
gacttggaga acaagatcaa ggacatcaac atcaaatcca atcttgagga aaacaatacc 540
atgaacgaat ccggcaaagt ggatagcaag tatgagctca ctggggacga gaaatgtggt 600
aaaagcctga agctcggcaa catcagcaat cagacaaacc aggaaaccat aacccaaagc 660
ctgtcagttg gagagattct gtgcattgac ctcgaaggga atgcaggaac aggctatctg 720
tgggtgttgc tgggcataca caaggatgag ccaatcataa accccgagaa cttcccaacc 780
aaactgacaa agaagtcttt cttttccgag gaaataagtg tgactcagcc aaagaagtac 840
aagatcgatg agcatgattc ttcaaagaat gtgaatcgcg aaatcgaaag ccctgaacag 900
aaggagtccg actcaaagcc caagaaacct cagatgcaac ttcttggagg accagatcgg 960
atgaggtcag tcatcaaagg acacaaacct ggcaaatatt acattgtgta ctcttactat 1020
cgaccgtttt ctcccacttc tggggcgaac actaaaatca tttacgtcac agtacagtga 1080
ctcgag 1086
<210> 29
<211> 355
<212> PRT
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> ICP
<400> 29
Met Ala Lys Ser Ile Thr Phe Phe Val Phe Asn Ile Cys Ser Ile Leu
1 5 10 15
Ala Leu Leu Ser His Cys Glu Asp Asn Asp Ile Tyr Ser Phe Asp Ile
20 25 30
Val Asn Glu Thr Asn Trp Leu Lys Ile Ala Lys Asn Ile Phe Lys Gly
35 40 45
Lys Ser Pro Ser Asn Phe Thr Ile Ile Pro Phe Asn Asn Thr Gly Ser
50 55 60
Ser Asn Asp Asn Glu Ser Asn Lys Glu Glu Ser Val Leu Leu Ile Arg
65 70 75 80
Lys Lys Ile Lys Ser Asn Lys Asn His Asp Ser Ser Ile Ile Ser Gly
85 90 95
Asp Thr Val Asn Gly Asp Ile Ser Asp Leu Asn Tyr Thr Ala Ser Asn
100 105 110
Phe Ser Asp Asn Ser Glu Asp Ile Glu Asp Asn Gln Lys Tyr Pro Thr
115 120 125
Thr Ser Tyr Asn Ser Phe Asn His Leu Asn Ser Asn Ile Ala Phe Asn
130 135 140
Glu Glu Ser Glu Tyr Ile Glu Ile Asn Ser Glu Ser Asp Leu Glu Asn
145 150 155 160
Lys Ile Lys Asp Ile Asn Ile Lys Ser Asn Leu Glu Glu Asn Asn Thr
165 170 175
Met Asn Glu Ser Gly Lys Val Asp Ser Lys Tyr Glu Leu Thr Gly Asp
180 185 190
Glu Lys Cys Gly Lys Ser Leu Lys Leu Gly Asn Ile Ser Asn Gln Thr
195 200 205
Asn Gln Glu Thr Ile Thr Gln Ser Leu Ser Val Gly Glu Ile Leu Cys
210 215 220
Ile Asp Leu Glu Gly Asn Ala Gly Thr Gly Tyr Leu Trp Val Leu Leu
225 230 235 240
Gly Ile His Lys Asp Glu Pro Ile Ile Asn Pro Glu Asn Phe Pro Thr
245 250 255
Lys Leu Thr Lys Lys Ser Phe Phe Ser Glu Glu Ile Ser Val Thr Gln
260 265 270
Pro Lys Lys Tyr Lys Ile Asp Glu His Asp Ser Ser Lys Asn Val Asn
275 280 285
Arg Glu Ile Glu Ser Pro Glu Gln Lys Glu Ser Asp Ser Lys Pro Lys
290 295 300
Lys Pro Gln Met Gln Leu Leu Gly Gly Pro Asp Arg Met Arg Ser Val
305 310 315 320
Ile Lys Gly His Lys Pro Gly Lys Tyr Tyr Ile Val Tyr Ser Tyr Tyr
325 330 335
Arg Pro Phe Ser Pro Thr Ser Gly Ala Asn Thr Lys Ile Ile Tyr Val
340 345 350
Thr Val Gln
355
<210> 30
<211> 354
<212> PRT
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> ICP
<400> 30
Met Lys Ser Ile Thr Phe Phe Val Phe Asn Ile Cys Ser Ile Leu Ala
1 5 10 15
Leu Leu Ser His Cys Glu Asp Asn Asp Ile Tyr Ser Phe Asp Ile Val
20 25 30
Asn Glu Thr Asn Trp Leu Lys Ile Ala Lys Asn Ile Phe Lys Gly Lys
35 40 45
Ser Pro Ser Asn Phe Thr Ile Ile Pro Phe Asn Asn Thr Gly Ser Ser
50 55 60
Asn Asp Asn Glu Ser Asn Lys Glu Glu Ser Val Leu Leu Ile Arg Lys
65 70 75 80
Lys Ile Lys Ser Asn Lys Asn His Asp Ser Ser Ile Ile Ser Gly Asp
85 90 95
Thr Val Asn Gly Asp Ile Ser Asp Leu Asn Tyr Thr Ala Ser Asn Phe
100 105 110
Ser Asp Asn Ser Glu Asp Ile Glu Asp Asn Gln Lys Tyr Pro Thr Thr
115 120 125
Ser Tyr Asn Ser Phe Asn His Leu Asn Ser Asn Ile Ala Phe Asn Glu
130 135 140
Glu Ser Glu Tyr Ile Glu Ile Asn Ser Glu Ser Asp Leu Glu Asn Lys
145 150 155 160
Ile Lys Asp Ile Asn Ile Lys Ser Asn Leu Glu Glu Asn Asn Thr Met
165 170 175
Asn Glu Ser Gly Lys Val Asp Ser Lys Tyr Glu Leu Thr Gly Asp Glu
180 185 190
Lys Cys Gly Lys Ser Leu Lys Leu Gly Asn Ile Ser Asn Gln Thr Asn
195 200 205
Gln Glu Thr Ile Thr Gln Ser Leu Ser Val Gly Glu Ile Leu Cys Ile
210 215 220
Asp Leu Glu Gly Asn Ala Gly Thr Gly Tyr Leu Trp Val Leu Leu Gly
225 230 235 240
Ile His Lys Asp Glu Pro Ile Ile Asn Pro Glu Asn Phe Pro Thr Lys
245 250 255
Leu Thr Lys Lys Ser Phe Phe Ser Glu Glu Ile Ser Val Thr Gln Pro
260 265 270
Lys Lys Tyr Lys Ile Asp Glu His Asp Ser Ser Lys Asn Val Asn Arg
275 280 285
Glu Ile Glu Ser Pro Glu Gln Lys Glu Ser Asp Ser Lys Pro Lys Lys
290 295 300
Pro Gln Met Gln Leu Leu Gly Gly Pro Asp Arg Met Arg Ser Val Ile
305 310 315 320
Lys Gly His Lys Pro Gly Lys Tyr Tyr Ile Val Tyr Ser Tyr Tyr Arg
325 330 335
Pro Phe Ser Pro Thr Ser Gly Ala Asn Thr Lys Ile Ile Tyr Val Thr
340 345 350
Val Gln
<210> 31
<211> 1251
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> PfICP人CO
<400> 31
gccaccatgg ctaatctgtt ggtgttcttt tgcttctttc tgttgtcttg catcgtacac 60
ctctctcgtt gttctgacaa caattcatat agctttgaga tcgtgaatcg gagcacatgg 120
ctcaacattg ctgaacggat attcaaaggc aatgcaccat tcaactttac tattataccg 180
tacaactatg tgaacaatag tactgaagag aataataaca aagactctgt ccttctgata 240
agcaagaatc tgaagaattc ctcaaaccca gtggacgaaa ataaccacat cattgactca 300
accaagaaga acacgtccaa taacaacaat aataacagca atatcgtcgg aatatacgaa 360
agtcaggtac atgaggagaa gattaaagag gacaatacaa gacaggataa tatcaacaag 420
aaggaaaacg agataatcaa caataaccat cagatccctg tgtccaacat cttttcagag 480
aacattgaca acaacaagaa ctacattgag agcaactaca agagcaccta taacaataat 540
cccgagttga ttcatagcac agatttcatt ggcagtaata acaaccacac tttcaatttc 600
ctgtctcgct ataacaactc agtgctgaac aacatgcaag ggaataccaa agttccaggg 660
aatgttcccg aactgaaagc ccgcattttc tccgaggaag aaaacaccga agtcgaaagc 720
gccgaaaaca atcacactaa cagtctgaat cctaacgagt cttgtgatca aatcatcaaa 780
ctgggcgata tcattaacag cgtcaatgag aagatcatca gcatcaatag tacggtgaat 840
aacgtgctct gcataaatct ggattccgtc aatggcaatg gcttcgtttg gacccttctt 900
ggggtacaca agaagaaacc cctgattgac ccctccaatt ttcccactaa aagggtgact 960
cagtcctacg tttcacctga catttccgtt acaaaccctg tgccaattcc aaagaacagc 1020
aacaccaaca aagatgacag catcaataat aaacaggatg gttcccagaa taacacaaca 1080
acgaatcact ttccgaagcc tagagagcaa ctcgtgggtg gatcttctat gctgatcagt 1140
aagatcaaac cccataaacc cggaaagtat ttcatcgtgt atagctacta cagaccattt 1200
gaccctacaa gggataccaa cacccgaatt gtggagctga atgtccagta a 1251
<210> 32
<211> 414
<212> PRT
<213> 恶性疟原虫（Plasmodium falciparum）
<220>
<221> misc_feature
<223> PfICP
<400> 32
Met Ala Asn Leu Leu Val Phe Phe Cys Phe Phe Leu Leu Ser Cys Ile
1 5 10 15
Val His Leu Ser Arg Cys Ser Asp Asn Asn Ser Tyr Ser Phe Glu Ile
20 25 30
Val Asn Arg Ser Thr Trp Leu Asn Ile Ala Glu Arg Ile Phe Lys Gly
35 40 45
Asn Ala Pro Phe Asn Phe Thr Ile Ile Pro Tyr Asn Tyr Val Asn Asn
50 55 60
Ser Thr Glu Glu Asn Asn Asn Lys Asp Ser Val Leu Leu Ile Ser Lys
65 70 75 80
Asn Leu Lys Asn Ser Ser Asn Pro Val Asp Glu Asn Asn His Ile Ile
85 90 95
Asp Ser Thr Lys Lys Asn Thr Ser Asn Asn Asn Asn Asn Asn Ser Asn
100 105 110
Ile Val Gly Ile Tyr Glu Ser Gln Val His Glu Glu Lys Ile Lys Glu
115 120 125
Asp Asn Thr Arg Gln Asp Asn Ile Asn Lys Lys Glu Asn Glu Ile Ile
130 135 140
Asn Asn Asn His Gln Ile Pro Val Ser Asn Ile Phe Ser Glu Asn Ile
145 150 155 160
Asp Asn Asn Lys Asn Tyr Ile Glu Ser Asn Tyr Lys Ser Thr Tyr Asn
165 170 175
Asn Asn Pro Glu Leu Ile His Ser Thr Asp Phe Ile Gly Ser Asn Asn
180 185 190
Asn His Thr Phe Asn Phe Leu Ser Arg Tyr Asn Asn Ser Val Leu Asn
195 200 205
Asn Met Gln Gly Asn Thr Lys Val Pro Gly Asn Val Pro Glu Leu Lys
210 215 220
Ala Arg Ile Phe Ser Glu Glu Glu Asn Thr Glu Val Glu Ser Ala Glu
225 230 235 240
Asn Asn His Thr Asn Ser Leu Asn Pro Asn Glu Ser Cys Asp Gln Ile
245 250 255
Ile Lys Leu Gly Asp Ile Ile Asn Ser Val Asn Glu Lys Ile Ile Ser
260 265 270
Ile Asn Ser Thr Val Asn Asn Val Leu Cys Ile Asn Leu Asp Ser Val
275 280 285
Asn Gly Asn Gly Phe Val Trp Thr Leu Leu Gly Val His Lys Lys Lys
290 295 300
Pro Leu Ile Asp Pro Ser Asn Phe Pro Thr Lys Arg Val Thr Gln Ser
305 310 315 320
Tyr Val Ser Pro Asp Ile Ser Val Thr Asn Pro Val Pro Ile Pro Lys
325 330 335
Asn Ser Asn Thr Asn Lys Asp Asp Ser Ile Asn Asn Lys Gln Asp Gly
340 345 350
Ser Gln Asn Asn Thr Thr Thr Asn His Phe Pro Lys Pro Arg Glu Gln
355 360 365
Leu Val Gly Gly Ser Ser Met Leu Ile Ser Lys Ile Lys Pro His Lys
370 375 380
Pro Gly Lys Tyr Phe Ile Val Tyr Ser Tyr Tyr Arg Pro Phe Asp Pro
385 390 395 400
Thr Arg Asp Thr Asn Thr Arg Ile Val Glu Leu Asn Val Gln
405 410
<210> 33
<211> 413
<212> PRT
<213> 恶性疟原虫（Plasmodium falciparum）
<220>
<221> misc_feature
<223> ICP
<400> 33
Met Asn Leu Leu Val Phe Phe Cys Phe Phe Leu Leu Ser Cys Ile Val
1 5 10 15
His Leu Ser Arg Cys Ser Asp Asn Asn Ser Tyr Ser Phe Glu Ile Val
20 25 30
Asn Arg Ser Thr Trp Leu Asn Ile Ala Glu Arg Ile Phe Lys Gly Asn
35 40 45
Ala Pro Phe Asn Phe Thr Ile Ile Pro Tyr Asn Tyr Val Asn Asn Ser
50 55 60
Thr Glu Glu Asn Asn Asn Lys Asp Ser Val Leu Leu Ile Ser Lys Asn
65 70 75 80
Leu Lys Asn Ser Ser Asn Pro Val Asp Glu Asn Asn His Ile Ile Asp
85 90 95
Ser Thr Lys Lys Asn Thr Ser Asn Asn Asn Asn Asn Asn Ser Asn Ile
100 105 110
Val Gly Ile Tyr Glu Ser Gln Val His Glu Glu Lys Ile Lys Glu Asp
115 120 125
Asn Thr Arg Gln Asp Asn Ile Asn Lys Lys Glu Asn Glu Ile Ile Asn
130 135 140
Asn Asn His Gln Ile Pro Val Ser Asn Ile Phe Ser Glu Asn Ile Asp
145 150 155 160
Asn Asn Lys Asn Tyr Ile Glu Ser Asn Tyr Lys Ser Thr Tyr Asn Asn
165 170 175
Asn Pro Glu Leu Ile His Ser Thr Asp Phe Ile Gly Ser Asn Asn Asn
180 185 190
His Thr Phe Asn Phe Leu Ser Arg Tyr Asn Asn Ser Val Leu Asn Asn
195 200 205
Met Gln Gly Asn Thr Lys Val Pro Gly Asn Val Pro Glu Leu Lys Ala
210 215 220
Arg Ile Phe Ser Glu Glu Glu Asn Thr Glu Val Glu Ser Ala Glu Asn
225 230 235 240
Asn His Thr Asn Ser Leu Asn Pro Asn Glu Ser Cys Asp Gln Ile Ile
245 250 255
Lys Leu Gly Asp Ile Ile Asn Ser Val Asn Glu Lys Ile Ile Ser Ile
260 265 270
Asn Ser Thr Val Asn Asn Val Leu Cys Ile Asn Leu Asp Ser Val Asn
275 280 285
Gly Asn Gly Phe Val Trp Thr Leu Leu Gly Val His Lys Lys Lys Pro
290 295 300
Leu Ile Asp Pro Ser Asn Phe Pro Thr Lys Arg Val Thr Gln Ser Tyr
305 310 315 320
Val Ser Pro Asp Ile Ser Val Thr Asn Pro Val Pro Ile Pro Lys Asn
325 330 335
Ser Asn Thr Asn Lys Asp Asp Ser Ile Asn Asn Lys Gln Asp Gly Ser
340 345 350
Gln Asn Asn Thr Thr Thr Asn His Phe Pro Lys Pro Arg Glu Gln Leu
355 360 365
Val Gly Gly Ser Ser Met Leu Ile Ser Lys Ile Lys Pro His Lys Pro
370 375 380
Gly Lys Tyr Phe Ile Val Tyr Ser Tyr Tyr Arg Pro Phe Asp Pro Thr
385 390 395 400
Arg Asp Thr Asn Thr Arg Ile Val Glu Leu Asn Val Gln
405 410
<210> 34
<211> 1104
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> PvICP人CO
<400> 34
gccaccatgg ctaagttgtc tagcctgttc tgcctggttg tgtgttctag tgttgcccac 60
ctctcttcct gtagtgatca aaacacttat agttttgata ttgttaatcg aaacacttgg 120
tacagcatag ccaagaaaat ctttcaaggc acgaccccct gtaatttcac tgtaatccct 180
agttcctatg tcaacaactc tgacggagtg tctacgagtg atgattccgt actgctcatc 240
cgcaaaaagc tcaaggatcc gagtgaagct ggccttgacg gatcttcagt ttctggttca 300
tccagttctg gaaacagtca ctccggttct gcaccttgtt gtgataaggg tacccccgct 360
aaagaggcag agctgaaatt ttctacaaag tttgagggcg atgactatgc taagctgaga 420
gattctctga gccttataga caagtcactc cgagaagagt caagctcaga ggaggacagt 480
aagatggaag atagtcaggt cggtgaagta actcatgaag agactatcac ctacaacatg 540
cccgaagaat atatgcccca gaacatttcc gaggtattga tcggtgccgc tgaagaggat 600
aggacatacg cgttgaaggg ggacgagccg tgtgatgtgt acttgaaact tggcgagata 660
atcaatggaa ctaatgaaaa gactatcgag tattctctcc aaaaaaataa gatactgtgc 720
gttcaactcg aagcaattgg gggaaatgga tacctctggg ctctcctcgg cgtacacaaa 780
gaaaaacccc aaatcaaccc agaggagttt ccacgaaaaa agatcacaaa atcttttttc 840
accaatgaga tatccgtcac gcagcctaaa gcagtgcaaa agaacaaatc taataatgga 900
ggtgagagca gttcaaactc acctggctat gggaaacccc ccgcaagcga acagctcggg 960
ggatttgtgg gtggcacatc catgcttcag agtatagtaa aggctcataa agagggcacc 1020
tttttcgtag tttatagcta ctaccgcccc ttcgacccta ccgccaacgc caacacgaag 1080
atactcaaac ttacggtttc atag 1104
<210> 35
<211> 365
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> PVICP 人CO + kozac
<220>
<221> misc_feature
<223> PvICP
<400> 35
Met Ala Lys Leu Ser Ser Leu Phe Cys Leu Val Val Cys Ser Ser Val
1 5 10 15
Ala His Leu Ser Ser Cys Ser Asp Gln Asn Thr Tyr Ser Phe Asp Ile
20 25 30
Val Asn Arg Asn Thr Trp Tyr Ser Ile Ala Lys Lys Ile Phe Gln Gly
35 40 45
Thr Thr Pro Cys Asn Phe Thr Val Ile Pro Ser Ser Tyr Val Asn Asn
50 55 60
Ser Asp Gly Val Ser Thr Ser Asp Asp Ser Val Leu Leu Ile Arg Lys
65 70 75 80
Lys Leu Lys Asp Pro Ser Glu Ala Gly Leu Asp Gly Ser Ser Val Ser
85 90 95
Gly Ser Ser Ser Ser Gly Asn Ser His Ser Gly Ser Ala Pro Cys Cys
100 105 110
Asp Lys Gly Thr Pro Ala Lys Glu Ala Glu Leu Lys Phe Ser Thr Lys
115 120 125
Phe Glu Gly Asp Asp Tyr Ala Lys Leu Arg Asp Ser Leu Ser Leu Ile
130 135 140
Asp Lys Ser Leu Arg Glu Glu Ser Ser Ser Glu Glu Asp Ser Lys Met
145 150 155 160
Glu Asp Ser Gln Val Gly Glu Val Thr His Glu Glu Thr Ile Thr Tyr
165 170 175
Asn Met Pro Glu Glu Tyr Met Pro Gln Asn Ile Ser Glu Val Leu Ile
180 185 190
Gly Ala Ala Glu Glu Asp Arg Thr Tyr Ala Leu Lys Gly Asp Glu Pro
195 200 205
Cys Asp Val Tyr Leu Lys Leu Gly Glu Ile Ile Asn Gly Thr Asn Glu
210 215 220
Lys Thr Ile Glu Tyr Ser Leu Gln Lys Asn Lys Ile Leu Cys Val Gln
225 230 235 240
Leu Glu Ala Ile Gly Gly Asn Gly Tyr Leu Trp Ala Leu Leu Gly Val
245 250 255
His Lys Glu Lys Pro Gln Ile Asn Pro Glu Glu Phe Pro Arg Lys Lys
260 265 270
Ile Thr Lys Ser Phe Phe Thr Asn Glu Ile Ser Val Thr Gln Pro Lys
275 280 285
Ala Val Gln Lys Asn Lys Ser Asn Asn Gly Gly Glu Ser Ser Ser Asn
290 295 300
Ser Pro Gly Tyr Gly Lys Pro Pro Ala Ser Glu Gln Leu Gly Gly Phe
305 310 315 320
Val Gly Gly Thr Ser Met Leu Gln Ser Ile Val Lys Ala His Lys Glu
325 330 335
Gly Thr Phe Phe Val Val Tyr Ser Tyr Tyr Arg Pro Phe Asp Pro Thr
340 345 350
Ala Asn Ala Asn Thr Lys Ile Leu Lys Leu Thr Val Ser
355 360 365
<210> 36
<211> 359
<212> PRT
<213> 间日疟原虫（Plasmodium vivax）
<220>
<221> misc_feature
<223> ICP
<400> 36
Met Lys Leu Ser Ser Leu Phe Cys Leu Val Val Cys Ser Ser Val Ala
1 5 10 15
His Leu Ser Ser Cys Ser Asp Gln Asn Thr Tyr Ser Phe Asp Ile Val
20 25 30
Asn Arg Asn Thr Trp Tyr Ser Ile Ala Lys Lys Ile Phe Gln Gly Thr
35 40 45
Thr Pro Cys Asn Phe Thr Val Ile Pro Ser Ser Tyr Val Asn Asn Ser
50 55 60
Asp Gly Val Ser Thr Ser Asp Asp Ser Val Leu Leu Ile Arg Lys Lys
65 70 75 80
Leu Lys Asp Pro Ser Glu Ala Gly Leu Asp Gly Ser Ser Val Ser Gly
85 90 95
Asn Ser His Ser Gly Ser Ala Pro Cys Cys Asp Lys Gly Thr Pro Ala
100 105 110
Lys Glu Ala Glu Leu Asn Phe Ser Thr Lys Phe Glu Gly Asp Asp Tyr
115 120 125
Ala Lys Leu Arg Asp Ser Leu Ser Leu Ile Asp Lys Ser Leu Arg Glu
130 135 140
Glu Ser Ser Ser Glu Glu Asp Ser Lys Met Glu Asp Ser Gln Val Gly
145 150 155 160
Glu Val Thr His Glu Glu Thr Ile Thr Tyr Asn Met Pro Glu Glu Tyr
165 170 175
Met Pro Gln Asn Ile Ser Glu Val Leu Ile Gly Ala Ala Glu Glu Asp
180 185 190
Arg Thr Tyr Ala Leu Lys Gly Asp Glu Pro Cys Asp Val Tyr Leu Lys
195 200 205
Leu Gly Glu Ile Ile Asn Gly Thr Asn Glu Lys Thr Ile Glu Tyr Ser
210 215 220
Leu Gln Lys Asn Lys Ile Leu Cys Val Gln Leu Glu Ala Ile Gly Gly
225 230 235 240
Asn Gly Tyr Leu Trp Ala Leu Leu Gly Val His Lys Glu Lys Pro Gln
245 250 255
Ile Asn Pro Glu Glu Phe Pro Arg Lys Lys Ile Thr Lys Ser Phe Phe
260 265 270
Thr Asn Glu Ile Ser Val Thr Gln Pro Lys Ala Val Gln Lys Asn Lys
275 280 285
Ser Asn Asn Gly Gly Glu Ser Ser Ser Asn Ser Pro Gly Tyr Gly Lys
290 295 300
Pro Pro Ala Ser Glu Gln Leu Gly Gly Phe Val Gly Gly Thr Ser Met
305 310 315 320
Leu Gln Ser Ile Val Lys Ala His Lys Glu Gly Thr Phe Phe Val Val
325 330 335
Tyr Ser Tyr Tyr Arg Pro Phe Asp Pro Thr Ala Asn Ala Asn Thr Lys
340 345 350
Ile Leu Lys Leu Thr Val Ser
355
<210> 37
<211> 2355
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> Falcilysin，Bergheilysin-A-CO
<400> 37
ggatccgcca ccatggctaa attgatgaag gttttgggtt acatcaacat tattaccaac 60
tgcgtgaatg gaatcctctg caaaggtgac aagaaaaggt actccatctt caccaataat 120
tacatctaca gcatatcaac cctcaataat tatagctttg ctgcaacaat gaacaaaatg 180
cccgcttggg ttaacgagaa atgtccagaa cacaaaagct acgacatcgt tgagaagcgc 240
tacaatgaga acctcaatct cacgtacaca gtctatgagc acaagaaggc caagactcag 300
gtcatagcac tggggtctaa cgatcctctc gacgctgagc aagcatttgg cttctacgtg 360
aaaaccctga cgcattcaga taaaggcatt ccgcacatac tggaacacac tgtcctgagt 420
ggctctaaga atttcaacta caaggactca atggggcttt tggagaaagg caccctgaac 480
acacacctga atgcctacac cttcaatgac aggaccatct acatggccgg gagtatgaac 540
aatagggatt tctttaacat tatggccgtc tacatggata gcgtgttcca gcctaacgta 600
ctggaaaaca aattcatctt ccagacagag ggatggacct atgaggtaga gaagctgaag 660
gaggaggaga agaacctcga cattcccaag attaaggact acaaggtgtc ttttaacgga 720
atcgtgtata atgagatgaa gggtgcgttt agcaatcctc tgcaggacct gtattatgaa 780
gtgatgagaa acatgttccc cgacaacgta cacagtaaca tctccggagg agatcctaaa 840
gaaataccaa atctgtcata tgaagagttt aaggagtttt actacaagaa ttacaatccg 900
aagaaaatca aagtgttctt tttctccaaa aataatccga cagagctgct caactttgtg 960
gacaactatc tctgtcagct ggacttcacg aaatatcggg atgatgctgt ggaacatgtc 1020
aattaccaag aataccgcaa aggcccattc tatattaaga agaaattcgc tgatcactca 1080
gaagagaaag aaaatcttgc ctccgtgagt tggcttctga atcccaagaa acacaagaat 1140
tccgatactg atctctctct ggagtctcct acagactatt tcgccttgct catcatcaat 1200
aatctgctta ctcataccag tgagagcgtc ctgtacaaag ccctgataga atccggattg 1260
gggaatagta ttgtagatcg agggctgaat gattccctgg ttcagtatgt gttcagcatc 1320
ggcctgaaag gcataaaaga gaagaacgag aagaacatct ccctggacaa ggtccactac 1380
gaggtggaaa agatcgttct tgaggcactg aaaaaagtgg tcaaagaagg tttcaataag 1440
tcagcagttg aggcagccat taacaatatt gagttcgtcc tgaaagaagc caatctcaag 1500
atctccaaat ctatagactt tgtgtttgaa atggccagca gactgaacta tggcaaagat 1560
ccactgctga tctttgagtt tgaaaagcat ctcaacgtag tgaaggacaa gatcaagaac 1620
gagcctaaat acctggagaa gtatgtggag aaacatcttc tgaacaatga tcatcgagtc 1680
gttattctgc tggaagggga tgaaaactat ggcaccgaac aggagaaact ggaaaaggac 1740
atgctgaaga agcggattga aagcttcact gagaaagaaa aggagaatat tatcacagac 1800
ttcgaaaatc ttacgaagta caagaacact gaggaatctc ccgaacatct ggacaagttt 1860
cccatcatta gcattagcga cctcaatgga aagactttgg agatccccgt gaaccctttc 1920
tttaccaacc tgaacaacga gaacaacatg cagcactata atgagacaaa gaacaaccaa 1980
accctggtca aagaaaacat ggaccgtttc attaacaaat acattctcaa caaggatgga 2040
aacgataaga acgacagcaa gaacgcggat gtgccaatgc tgatttatga aatcccaaca 2100
tctggcatat tgtatctgca gtttatcttc tctctggata accttacact ggaagaattg 2160
tcctacctga atctgtttaa aagcttgata ctggagaaca agactaacaa gagatcaagt 2220
gaagagtttg tgatcttgcg ggagaagaat attgggaata tgatgaccaa tgttgctctt 2280
cttagcacat ccgatcgcct caatgtgact gacaaatata atgcgaaagg tttctttaac 2340
tttgagtaac tcgag 2355
<210> 38
<211> 778
<212> PRT
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> Falcilysin，Bergheilysin-A
<400> 38
Met Ala Lys Leu Met Lys Val Leu Gly Tyr Ile Asn Ile Ile Thr Asn
1 5 10 15
Cys Val Asn Gly Ile Leu Cys Lys Gly Asp Lys Lys Arg Tyr Ser Ile
20 25 30
Phe Thr Asn Asn Tyr Ile Tyr Ser Ile Ser Thr Leu Asn Asn Tyr Ser
35 40 45
Phe Ala Ala Thr Met Asn Lys Met Pro Ala Trp Val Asn Glu Lys Cys
50 55 60
Pro Glu His Lys Ser Tyr Asp Ile Val Glu Lys Arg Tyr Asn Glu Asn
65 70 75 80
Leu Asn Leu Thr Tyr Thr Val Tyr Glu His Lys Lys Ala Lys Thr Gln
85 90 95
Val Ile Ala Leu Gly Ser Asn Asp Pro Leu Asp Ala Glu Gln Ala Phe
100 105 110
Gly Phe Tyr Val Lys Thr Leu Thr His Ser Asp Lys Gly Ile Pro His
115 120 125
Ile Leu Glu His Thr Val Leu Ser Gly Ser Lys Asn Phe Asn Tyr Lys
130 135 140
Asp Ser Met Gly Leu Leu Glu Lys Gly Thr Leu Asn Thr His Leu Asn
145 150 155 160
Ala Tyr Thr Phe Asn Asp Arg Thr Ile Tyr Met Ala Gly Ser Met Asn
165 170 175
Asn Arg Asp Phe Phe Asn Ile Met Ala Val Tyr Met Asp Ser Val Phe
180 185 190
Gln Pro Asn Val Leu Glu Asn Lys Phe Ile Phe Gln Thr Glu Gly Trp
195 200 205
Thr Tyr Glu Val Glu Lys Leu Lys Glu Glu Glu Lys Asn Leu Asp Ile
210 215 220
Pro Lys Ile Lys Asp Tyr Lys Val Ser Phe Asn Gly Ile Val Tyr Asn
225 230 235 240
Glu Met Lys Gly Ala Phe Ser Asn Pro Leu Gln Asp Leu Tyr Tyr Glu
245 250 255
Val Met Arg Asn Met Phe Pro Asp Asn Val His Ser Asn Ile Ser Gly
260 265 270
Gly Asp Pro Lys Glu Ile Pro Asn Leu Ser Tyr Glu Glu Phe Lys Glu
275 280 285
Phe Tyr Tyr Lys Asn Tyr Asn Pro Lys Lys Ile Lys Val Phe Phe Phe
290 295 300
Ser Lys Asn Asn Pro Thr Glu Leu Leu Asn Phe Val Asp Asn Tyr Leu
305 310 315 320
Cys Gln Leu Asp Phe Thr Lys Tyr Arg Asp Asp Ala Val Glu His Val
325 330 335
Asn Tyr Gln Glu Tyr Arg Lys Gly Pro Phe Tyr Ile Lys Lys Lys Phe
340 345 350
Ala Asp His Ser Glu Glu Lys Glu Asn Leu Ala Ser Val Ser Trp Leu
355 360 365
Leu Asn Pro Lys Lys His Lys Asn Ser Asp Thr Asp Leu Ser Leu Glu
370 375 380
Ser Pro Thr Asp Tyr Phe Ala Leu Leu Ile Ile Asn Asn Leu Leu Thr
385 390 395 400
His Thr Ser Glu Ser Val Leu Tyr Lys Ala Leu Ile Glu Ser Gly Leu
405 410 415
Gly Asn Ser Ile Val Asp Arg Gly Leu Asn Asp Ser Leu Val Gln Tyr
420 425 430
Val Phe Ser Ile Gly Leu Lys Gly Ile Lys Glu Lys Asn Glu Lys Asn
435 440 445
Ile Ser Leu Asp Lys Val His Tyr Glu Val Glu Lys Ile Val Leu Glu
450 455 460
Ala Leu Lys Lys Val Val Lys Glu Gly Phe Asn Lys Ser Ala Val Glu
465 470 475 480
Ala Ala Ile Asn Asn Ile Glu Phe Val Leu Lys Glu Ala Asn Leu Lys
485 490 495
Ile Ser Lys Ser Ile Asp Phe Val Phe Glu Met Ala Ser Arg Leu Asn
500 505 510
Tyr Gly Lys Asp Pro Leu Leu Ile Phe Glu Phe Glu Lys His Leu Asn
515 520 525
Val Val Lys Asp Lys Ile Lys Asn Glu Pro Lys Tyr Leu Glu Lys Tyr
530 535 540
Val Glu Lys His Leu Leu Asn Asn Asp His Arg Val Val Ile Leu Leu
545 550 555 560
Glu Gly Asp Glu Asn Tyr Gly Thr Glu Gln Glu Lys Leu Glu Lys Asp
565 570 575
Met Leu Lys Lys Arg Ile Glu Ser Phe Thr Glu Lys Glu Lys Glu Asn
580 585 590
Ile Ile Thr Asp Phe Glu Asn Leu Thr Lys Tyr Lys Asn Thr Glu Glu
595 600 605
Ser Pro Glu His Leu Asp Lys Phe Pro Ile Ile Ser Ile Ser Asp Leu
610 615 620
Asn Gly Lys Thr Leu Glu Ile Pro Val Asn Pro Phe Phe Thr Asn Leu
625 630 635 640
Asn Asn Glu Asn Asn Met Gln His Tyr Asn Glu Thr Lys Asn Asn Gln
645 650 655
Thr Leu Val Lys Glu Asn Met Asp Arg Phe Ile Asn Lys Tyr Ile Leu
660 665 670
Asn Lys Asp Gly Asn Asp Lys Asn Asp Ser Lys Asn Ala Asp Val Pro
675 680 685
Met Leu Ile Tyr Glu Ile Pro Thr Ser Gly Ile Leu Tyr Leu Gln Phe
690 695 700
Ile Phe Ser Leu Asp Asn Leu Thr Leu Glu Glu Leu Ser Tyr Leu Asn
705 710 715 720
Leu Phe Lys Ser Leu Ile Leu Glu Asn Lys Thr Asn Lys Arg Ser Ser
725 730 735
Glu Glu Phe Val Ile Leu Arg Glu Lys Asn Ile Gly Asn Met Met Thr
740 745 750
Asn Val Ala Leu Leu Ser Thr Ser Asp Arg Leu Asn Val Thr Asp Lys
755 760 765
Tyr Asn Ala Lys Gly Phe Phe Asn Phe Glu
770 775
<210> 39
<211> 1149
<212> PRT
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> Falcilysin，Bergheilysin-A
<400> 39
Met Lys Leu Met Lys Val Leu Gly Tyr Ile Asn Ile Ile Thr Asn Cys
1 5 10 15
Val Asn Gly Ile Leu Cys Lys Gly Asp Lys Lys Arg Tyr Ser Ile Phe
20 25 30
Thr Asn Asn Tyr Ile Tyr Ser Ile Ser Thr Leu Asn Asn Tyr Ser Phe
35 40 45
Ala Ala Thr Met Asn Lys Met Pro Ala Trp Val Asn Glu Lys Cys Pro
50 55 60
Glu His Lys Ser Tyr Asp Ile Val Glu Lys Arg Tyr Asn Glu Asn Leu
65 70 75 80
Asn Leu Thr Tyr Thr Val Tyr Glu His Lys Lys Ala Lys Thr Gln Val
85 90 95
Ile Ala Leu Gly Ser Asn Asp Pro Leu Asp Ala Glu Gln Ala Phe Gly
100 105 110
Phe Tyr Val Lys Thr Leu Thr His Ser Asp Lys Gly Ile Pro His Ile
115 120 125
Leu Glu His Thr Val Leu Ser Gly Ser Lys Asn Phe Asn Tyr Lys Asp
130 135 140
Ser Met Gly Leu Leu Glu Lys Gly Thr Leu Asn Thr His Leu Asn Ala
145 150 155 160
Tyr Thr Phe Asn Asp Arg Thr Ile Tyr Met Ala Gly Ser Met Asn Asn
165 170 175
Arg Asp Phe Phe Asn Ile Met Ala Val Tyr Met Asp Ser Val Phe Gln
180 185 190
Pro Asn Val Leu Glu Asn Lys Phe Ile Phe Gln Thr Glu Gly Trp Thr
195 200 205
Tyr Glu Val Glu Lys Leu Lys Glu Glu Glu Lys Asn Leu Asp Ile Pro
210 215 220
Lys Ile Lys Asp Tyr Lys Val Ser Phe Asn Gly Ile Val Tyr Asn Glu
225 230 235 240
Met Lys Gly Ala Phe Ser Asn Pro Leu Gln Asp Leu Tyr Tyr Glu Val
245 250 255
Met Arg Asn Met Phe Pro Asp Asn Val His Ser Asn Ile Ser Gly Gly
260 265 270
Asp Pro Lys Glu Ile Pro Asn Leu Ser Tyr Glu Glu Phe Lys Glu Phe
275 280 285
Tyr Tyr Lys Asn Tyr Asn Pro Lys Lys Ile Lys Val Phe Phe Phe Ser
290 295 300
Lys Asn Asn Pro Thr Glu Leu Leu Asn Phe Val Asp Asn Tyr Leu Cys
305 310 315 320
Gln Leu Asp Phe Thr Lys Tyr Arg Asp Asp Ala Val Glu His Val Asn
325 330 335
Tyr Gln Glu Tyr Arg Lys Gly Pro Phe Tyr Ile Lys Lys Lys Phe Ala
340 345 350
Asp His Ser Glu Glu Lys Glu Asn Leu Ala Ser Val Ser Trp Leu Leu
355 360 365
Asn Pro Lys Lys His Lys Asn Ser Asp Thr Asp Leu Ser Leu Glu Ser
370 375 380
Pro Thr Asp Tyr Phe Ala Leu Leu Ile Ile Asn Asn Leu Leu Thr His
385 390 395 400
Thr Ser Glu Ser Val Leu Tyr Lys Ala Leu Ile Glu Ser Gly Leu Gly
405 410 415
Asn Ser Ile Val Asp Arg Gly Leu Asn Asp Ser Leu Val Gln Tyr Val
420 425 430
Phe Ser Ile Gly Leu Lys Gly Ile Lys Glu Lys Asn Glu Lys Asn Ile
435 440 445
Ser Leu Asp Lys Val His Tyr Glu Val Glu Lys Ile Val Leu Glu Ala
450 455 460
Leu Lys Lys Val Val Lys Glu Gly Phe Asn Lys Ser Ala Val Glu Ala
465 470 475 480
Ala Ile Asn Asn Ile Glu Phe Val Leu Lys Glu Ala Asn Leu Lys Ile
485 490 495
Ser Lys Ser Ile Asp Phe Val Phe Glu Met Ala Ser Arg Leu Asn Tyr
500 505 510
Gly Lys Asp Pro Leu Leu Ile Phe Glu Phe Glu Lys His Leu Asn Val
515 520 525
Val Lys Asp Lys Ile Lys Asn Glu Pro Lys Tyr Leu Glu Lys Tyr Val
530 535 540
Glu Lys His Leu Leu Asn Asn Asp His Arg Val Val Ile Leu Leu Glu
545 550 555 560
Gly Asp Glu Asn Tyr Gly Thr Glu Gln Glu Lys Leu Glu Lys Asp Met
565 570 575
Leu Lys Lys Arg Ile Glu Ser Phe Thr Glu Lys Glu Lys Glu Asn Ile
580 585 590
Ile Thr Asp Phe Glu Asn Leu Thr Lys Tyr Lys Asn Thr Glu Glu Ser
595 600 605
Pro Glu His Leu Asp Lys Phe Pro Ile Ile Ser Ile Ser Asp Leu Asn
610 615 620
Gly Lys Thr Leu Glu Ile Pro Val Asn Pro Phe Phe Thr Asn Leu Asn
625 630 635 640
Asn Glu Asn Asn Met Gln His Tyr Asn Glu Thr Lys Asn Asn Gln Thr
645 650 655
Leu Val Lys Glu Asn Met Asp Arg Phe Ile Asn Lys Tyr Ile Leu Asn
660 665 670
Lys Asp Gly Asn Asp Lys Asn Asp Ser Lys Asn Ala Asp Val Pro Met
675 680 685
Leu Ile Tyr Glu Ile Pro Thr Ser Gly Ile Leu Tyr Leu Gln Phe Ile
690 695 700
Phe Ser Leu Asp Asn Leu Thr Leu Glu Glu Leu Ser Tyr Leu Asn Leu
705 710 715 720
Phe Lys Ser Leu Ile Leu Glu Asn Lys Thr Asn Lys Arg Ser Ser Glu
725 730 735
Glu Phe Val Ile Leu Arg Glu Lys Asn Ile Gly Asn Met Met Thr Asn
740 745 750
Val Ala Leu Leu Ser Thr Ser Asp Arg Leu Asn Val Thr Asp Lys Tyr
755 760 765
Asn Ala Lys Gly Phe Phe Asn Phe Glu Met His Met Leu Ser His Lys
770 775 780
Cys Asn Asp Ala Leu Glu Ile Ala Leu Glu Ala Leu Lys Glu Ser Asp
785 790 795 800
Phe Ser Asn Lys Lys Lys Val Ile Glu Ile Leu Lys Arg Lys Ile Asn
805 810 815
Gly Met Lys Thr Thr Phe Ala Ser Lys Gly His Ser Ile Leu Ile Lys
820 825 830
Tyr Val Lys Ser Arg Ile Asn Ser Lys Tyr Tyr Ala Tyr Asp Leu Ile
835 840 845
His Gly Tyr Asp Asn Tyr Leu Lys Leu Gln Glu Gln Leu Lys Leu Ala
850 855 860
Glu Thr Asn Tyr Glu Ser Leu Glu Ala Ile Leu Asn Arg Ile Arg Lys
865 870 875 880
Lys Ile Phe Lys Arg Asn Asn Leu Ile Met Asn Val Thr Val Asp Pro
885 890 895
Gly Thr Ile Asp Gln Leu Phe Ala Lys Ser Lys Asn Ser Phe Asn Asn
900 905 910
Leu Leu Ser Tyr Phe Asp Glu Asn Glu Ser Tyr Cys Ser Lys Asn Asp
915 920 925
Ser Phe Asn Lys Val Val Gly Trp Asn Lys Glu Ile Gln Glu Lys Lys
930 935 940
Leu Leu Glu Gly Glu Glu Val Lys Lys Glu Leu Leu Val Val Pro Thr
945 950 955 960
Phe Val Asn Ser Val Ser Met Ser Gly Val Leu Phe Asn Lys Gly Glu
965 970 975
Tyr Leu Asp Pro Ser Phe Thr Val Ile Val Ala Ala Leu Lys Asn Ser
980 985 990
Tyr Leu Trp Glu Thr Val Arg Gly Leu Asn Gly Ala Tyr Gly Val Phe
995 1000 1005
Ala Asp Ile Glu Tyr Asp Gly Thr Val Val Phe Leu Ser Ala Arg
1010 1015 1020
Asp Pro Asn Leu Glu Lys Thr Leu Gln Thr Phe Arg Glu Ala Ala
1025 1030 1035
Gln Gly Leu Arg Lys Met Ala Asp Val Met Thr Lys Asn Asp Leu
1040 1045 1050
Leu Arg Tyr Ile Ile Asn Ala Ile Gly Thr Ile Asp Arg Pro Arg
1055 1060 1065
Arg Gly Val Glu Leu Ser Lys Leu Ser Phe Ser Arg Ile Ile Ser
1070 1075 1080
Asn Glu Thr Glu Gln Asp Arg Ile Glu Phe Arg Asn Arg Val Met
1085 1090 1095
Asn Thr Lys Lys Glu Asp Phe Tyr Lys Phe Ala Asp Leu Leu Glu
1100 1105 1110
Lys Lys Val Lys Glu Phe Glu Lys Asn Val Val Ile Ile Thr Ser
1115 1120 1125
Lys Glu Lys Ala Asn Glu Tyr Ile Asn Asn Val Asp Asn Asp Phe
1130 1135 1140
Lys Lys Ile Leu Ile Glu
1145
<210> 40
<211> 3591
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> PfFalcilysin 人 CO
<400> 40
gccaccatgg ctaatcttac caaacttatg aaagtaatag gatatataaa cattataact 60
aattgtgtac aatcttttac taacagagcg gacaagaaga ggtacaatgt ttttgcaaag 120
tctttcatca atacgataaa cacgaacctg tacaccttca aagcggtgat gtcaaaaacg 180
cctgaatgga tccatgagaa atctcccaaa cacaacagtt atgacataat tgagaaacgc 240
tataatgaag agttcaaaat gacatacacg gtttatcaac ataaaaaagc aaagacgcag 300
gtaatttcac tcggtacgaa cgacccactt gatgtcgaac aggcctttgc cttctatgta 360
aagactctga cccactctgg gaaggggatt ccccacatcc tcgaacacag tgtcctctca 420
gggagtaaaa attacaacta caaaaattcc attggactcc ttgagaaggg aactctgcac 480
acccacctta acgcatacac cttcaatgac cgaactgtgt acatggccgg ttctatgaat 540
aacaaagact tcttcaatat aatgggcgtc tatatggaca gtgtcttcca acctaatgta 600
ctggaaaaca aatacatatt cgagacggag ggatggactt acgaggtgga aaagctgaaa 660
gaggatgaga agggaaaagc tgagattcca cagatgaaag attataaagt atctttcaac 720
ggaattgttt ataatgaaat gaagggtgcc ttgtcttccc cgttggaaga tctttaccat 780
gaagagatga agtatatgtt cccagacaac gtccactcta ataacagtgg cggagaccca 840
aaagagatca caaacttgac ctatgaggag ttcaaggagt tctattacaa aaactataat 900
ccaaaaaaag ttaaagtgtt cttcttttca aagaacaacc caacggagct tctcaatttc 960
gtagaccagt acctcggaca gctggactac agcaaatacc gcgacgatgc tgttgaaagt 1020
gttgaatacc aaacgtataa aaaaggacct ttctacatta aaaaaaagta tggggaccat 1080
agcgaagaga aggagaatct tgtttccgta gcgtggcttc tgaaccccaa ggttgacaag 1140
actaacaatc ataataataa ccatagtaat aaccaatcta gcgaaaataa tggttactcc 1200
aacggctccc actctagcga tttgtccttg gagaatccca cggactattt cgtgctcttg 1260
attatcaata acctccttat acatacccca gaaagcgtcc tgtacaaggc cctcacagat 1320
tgcgggctgg gaaataatgt aattgatagg ggtttgaatg attcacttgt ccaatacatt 1380
ttcagtattg ggctgaaggg aatcaaacgc aacaatgaaa aaattaaaaa cttcgataaa 1440
gtgcactatg aagtagaaga cgtaattatg aatgctctca aaaaagtggt caaggaggga 1500
ttcaataaat ccgccgtaga agccagcata aataatatcg agtttatcct gaaagaagcc 1560
aatttgaaaa cttcaaaatc tatagatttc gtttttgaga tgacttccaa gctcaattat 1620
aatagggatc cactgctgat cttcgagttt gagaaatatc ttaatattgt gaagaataag 1680
attaagaacg aacctatgta tttggagaag tttgttgaaa aacacttcat caacaacgca 1740
catcggtcag tgatccttct tgagggggac gagaactatg cacaggaaca ggaaaacctt 1800
gagaaacaag aactgaagaa acgcatagag aacttcaatg agcaggagaa agagcaagtc 1860
attaagaact tcgaggagct gtccaagtac aagaacgcgg aagagagccc ggaacacttg 1920
aacaagtttc caataatctc catttccgac ctcaataaga aaacactgga agtcccagtt 1980
aacgtctact tcacgaacat caacgaaaac aacaatataa tggagacata taacaagctg 2040
aagacaaatg agcacatgct taaggacaac atggacgtgt ttctcaaaaa atacgttctc 2100
aaaaacgata agcacaacac caacaataat aacaacaata ataataatat ggactactct 2160
ttcaccgaaa ctaaatatga aggaaatgtg ccaatccttg tgtacgaaat gccgacgact 2220
ggaatagtct atttgcagtt cgttttctcc ctcgatcacc tgaccgtaga cgagctcgcc 2280
tatttgaatt tgttcaagac acttatcttg gagaacaaga caaacaaacg ctccagtgag 2340
gatttcgtca ttttgagaga aaaaaatatt gggtcaatgt cagcgaatgt ggcgctctac 2400
agcaaggacg accacctgaa cgtaaccgac aagtataacg cgcaagcact cttcaatctg 2460
gaaatgcacg tactttctca taaatgcaac gatgcgctga acattgccct tgaagctgtt 2520
aaagaatctg acttcagtaa taaaaaaaag gtcatagata tccttaagag gaagattaat 2580
ggaatgaaaa ctacgttctc agaaaaagga tatgctatac tgatgaaata cgttaaagcc 2640
catcttaata gcaagcatta cgcccataac ataatttatg ggtacgaaaa ctatttgaag 2700
ctgcaagagc aattggagct tgcagaaaac gattttaaaa cattggagaa tattttggtg 2760
agaataagaa acaaaatctt taacaaaaaa aacttgatgg tcagcgtgac gtccgactat 2820
ggtgctctca aacacctgtt tgtgaatagc aacgaatcct tgaaaaacct tgtaagttac 2880
ttcgaggaaa acgataagta tatcaacgat atgcagaata aagtgaatga ccctaccgtc 2940
atggggtgga atgaagaaat caagtctaaa aagctgttcg acgaggagaa ggtgaaaaag 3000
gagttctttg tcttgccgac ctttgtcaac agcgtctcaa tgagtggtat cctgttcaag 3060
ccgggcgaat atctcgaccc gagtttcact gtcatcgttg cggccctgaa aaatagctat 3120
ttgtgggaca cagttcgagg acttaacggg gcttacggtg tattcgctga tatcgaatac 3180
gatggctctg tagtatttct ttcagctcga gaccccaatc tcgaaaagac cctcgccact 3240
ttccgagaat ctgctaaagg attgcgcaaa atggctgaca ccatgacaga gaacgatttg 3300
cttcggtata ttattaacac aattggcacg atcgacaagc ctcgacgagg gatagaactt 3360
tctaagctgt cattccttag acttatttcc aacgagtcag agcaagatcg ggtggagttc 3420
cggaaacgga taatgaacac aaaaaaggaa gatttctata aattcgcaga tcttcttgag 3480
agcaaagtaa atgaattcga aaaaaatatc gttatcataa caacaaaaga aaaggccaac 3540
gaatatatag caaacgtaga tggcgagttt aagaaagtct tgatcgagtg a 3591
<210> 41
<211> 1194
<212> PRT
<213> 恶性疟原虫（Plasmodium falciparum）
<220>
<221> misc_feature
<223> PfFalcilysin
<400> 41
Met Ala Asn Leu Thr Lys Leu Met Lys Val Ile Gly Tyr Ile Asn Ile
1 5 10 15
Ile Thr Asn Cys Val Gln Ser Phe Thr Asn Arg Ala Asp Lys Lys Arg
20 25 30
Tyr Asn Val Phe Ala Lys Ser Phe Ile Asn Thr Ile Asn Thr Asn Leu
35 40 45
Tyr Thr Phe Lys Ala Val Met Ser Lys Thr Pro Glu Trp Ile His Glu
50 55 60
Lys Ser Pro Lys His Asn Ser Tyr Asp Ile Ile Glu Lys Arg Tyr Asn
65 70 75 80
Glu Glu Phe Lys Met Thr Tyr Thr Val Tyr Gln His Lys Lys Ala Lys
85 90 95
Thr Gln Val Ile Ser Leu Gly Thr Asn Asp Pro Leu Asp Val Glu Gln
100 105 110
Ala Phe Ala Phe Tyr Val Lys Thr Leu Thr His Ser Gly Lys Gly Ile
115 120 125
Pro His Ile Leu Glu His Ser Val Leu Ser Gly Ser Lys Asn Tyr Asn
130 135 140
Tyr Lys Asn Ser Ile Gly Leu Leu Glu Lys Gly Thr Leu His Thr His
145 150 155 160
Leu Asn Ala Tyr Thr Phe Asn Asp Arg Thr Val Tyr Met Ala Gly Ser
165 170 175
Met Asn Asn Lys Asp Phe Phe Asn Ile Met Gly Val Tyr Met Asp Ser
180 185 190
Val Phe Gln Pro Asn Val Leu Glu Asn Lys Tyr Ile Phe Glu Thr Glu
195 200 205
Gly Trp Thr Tyr Glu Val Glu Lys Leu Lys Glu Asp Glu Lys Gly Lys
210 215 220
Ala Glu Ile Pro Gln Met Lys Asp Tyr Lys Val Ser Phe Asn Gly Ile
225 230 235 240
Val Tyr Asn Glu Met Lys Gly Ala Leu Ser Ser Pro Leu Glu Asp Leu
245 250 255
Tyr His Glu Glu Met Lys Tyr Met Phe Pro Asp Asn Val His Ser Asn
260 265 270
Asn Ser Gly Gly Asp Pro Lys Glu Ile Thr Asn Leu Thr Tyr Glu Glu
275 280 285
Phe Lys Glu Phe Tyr Tyr Lys Asn Tyr Asn Pro Lys Lys Val Lys Val
290 295 300
Phe Phe Phe Ser Lys Asn Asn Pro Thr Glu Leu Leu Asn Phe Val Asp
305 310 315 320
Gln Tyr Leu Gly Gln Leu Asp Tyr Ser Lys Tyr Arg Asp Asp Ala Val
325 330 335
Glu Ser Val Glu Tyr Gln Thr Tyr Lys Lys Gly Pro Phe Tyr Ile Lys
340 345 350
Lys Lys Tyr Gly Asp His Ser Glu Glu Lys Glu Asn Leu Val Ser Val
355 360 365
Ala Trp Leu Leu Asn Pro Lys Val Asp Lys Thr Asn Asn His Asn Asn
370 375 380
Asn His Ser Asn Asn Gln Ser Ser Glu Asn Asn Gly Tyr Ser Asn Gly
385 390 395 400
Ser His Ser Ser Asp Leu Ser Leu Glu Asn Pro Thr Asp Tyr Phe Val
405 410 415
Leu Leu Ile Ile Asn Asn Leu Leu Ile His Thr Pro Glu Ser Val Leu
420 425 430
Tyr Lys Ala Leu Thr Asp Cys Gly Leu Gly Asn Asn Val Ile Asp Arg
435 440 445
Gly Leu Asn Asp Ser Leu Val Gln Tyr Ile Phe Ser Ile Gly Leu Lys
450 455 460
Gly Ile Lys Arg Asn Asn Glu Lys Ile Lys Asn Phe Asp Lys Val His
465 470 475 480
Tyr Glu Val Glu Asp Val Ile Met Asn Ala Leu Lys Lys Val Val Lys
485 490 495
Glu Gly Phe Asn Lys Ser Ala Val Glu Ala Ser Ile Asn Asn Ile Glu
500 505 510
Phe Ile Leu Lys Glu Ala Asn Leu Lys Thr Ser Lys Ser Ile Asp Phe
515 520 525
Val Phe Glu Met Thr Ser Lys Leu Asn Tyr Asn Arg Asp Pro Leu Leu
530 535 540
Ile Phe Glu Phe Glu Lys Tyr Leu Asn Ile Val Lys Asn Lys Ile Lys
545 550 555 560
Asn Glu Pro Met Tyr Leu Glu Lys Phe Val Glu Lys His Phe Ile Asn
565 570 575
Asn Ala His Arg Ser Val Ile Leu Leu Glu Gly Asp Glu Asn Tyr Ala
580 585 590
Gln Glu Gln Glu Asn Leu Glu Lys Gln Glu Leu Lys Lys Arg Ile Glu
595 600 605
Asn Phe Asn Glu Gln Glu Lys Glu Gln Val Ile Lys Asn Phe Glu Glu
610 615 620
Leu Ser Lys Tyr Lys Asn Ala Glu Glu Ser Pro Glu His Leu Asn Lys
625 630 635 640
Phe Pro Ile Ile Ser Ile Ser Asp Leu Asn Lys Lys Thr Leu Glu Val
645 650 655
Pro Val Asn Val Tyr Phe Thr Asn Ile Asn Glu Asn Asn Asn Ile Met
660 665 670
Glu Thr Tyr Asn Lys Leu Lys Thr Asn Glu His Met Leu Lys Asp Asn
675 680 685
Met Asp Val Phe Leu Lys Lys Tyr Val Leu Lys Asn Asp Lys His Asn
690 695 700
Thr Asn Asn Asn Asn Asn Asn Asn Asn Asn Met Asp Tyr Ser Phe Thr
705 710 715 720
Glu Thr Lys Tyr Glu Gly Asn Val Pro Ile Leu Val Tyr Glu Met Pro
725 730 735
Thr Thr Gly Ile Val Tyr Leu Gln Phe Val Phe Ser Leu Asp His Leu
740 745 750
Thr Val Asp Glu Leu Ala Tyr Leu Asn Leu Phe Lys Thr Leu Ile Leu
755 760 765
Glu Asn Lys Thr Asn Lys Arg Ser Ser Glu Asp Phe Val Ile Leu Arg
770 775 780
Glu Lys Asn Ile Gly Ser Met Ser Ala Asn Val Ala Leu Tyr Ser Lys
785 790 795 800
Asp Asp His Leu Asn Val Thr Asp Lys Tyr Asn Ala Gln Ala Leu Phe
805 810 815
Asn Leu Glu Met His Val Leu Ser His Lys Cys Asn Asp Ala Leu Asn
820 825 830
Ile Ala Leu Glu Ala Val Lys Glu Ser Asp Phe Ser Asn Lys Lys Lys
835 840 845
Val Ile Asp Ile Leu Lys Arg Lys Ile Asn Gly Met Lys Thr Thr Phe
850 855 860
Ser Glu Lys Gly Tyr Ala Ile Leu Met Lys Tyr Val Lys Ala His Leu
865 870 875 880
Asn Ser Lys His Tyr Ala His Asn Ile Ile Tyr Gly Tyr Glu Asn Tyr
885 890 895
Leu Lys Leu Gln Glu Gln Leu Glu Leu Ala Glu Asn Asp Phe Lys Thr
900 905 910
Leu Glu Asn Ile Leu Val Arg Ile Arg Asn Lys Ile Phe Asn Lys Lys
915 920 925
Asn Leu Met Val Ser Val Thr Ser Asp Tyr Gly Ala Leu Lys His Leu
930 935 940
Phe Val Asn Ser Asn Glu Ser Leu Lys Asn Leu Val Ser Tyr Phe Glu
945 950 955 960
Glu Asn Asp Lys Tyr Ile Asn Asp Met Gln Asn Lys Val Asn Asp Pro
965 970 975
Thr Val Met Gly Trp Asn Glu Glu Ile Lys Ser Lys Lys Leu Phe Asp
980 985 990
Glu Glu Lys Val Lys Lys Glu Phe Phe Val Leu Pro Thr Phe Val Asn
995 1000 1005
Ser Val Ser Met Ser Gly Ile Leu Phe Lys Pro Gly Glu Tyr Leu
1010 1015 1020
Asp Pro Ser Phe Thr Val Ile Val Ala Ala Leu Lys Asn Ser Tyr
1025 1030 1035
Leu Trp Asp Thr Val Arg Gly Leu Asn Gly Ala Tyr Gly Val Phe
1040 1045 1050
Ala Asp Ile Glu Tyr Asp Gly Ser Val Val Phe Leu Ser Ala Arg
1055 1060 1065
Asp Pro Asn Leu Glu Lys Thr Leu Ala Thr Phe Arg Glu Ser Ala
1070 1075 1080
Lys Gly Leu Arg Lys Met Ala Asp Thr Met Thr Glu Asn Asp Leu
1085 1090 1095
Leu Arg Tyr Ile Ile Asn Thr Ile Gly Thr Ile Asp Lys Pro Arg
1100 1105 1110
Arg Gly Ile Glu Leu Ser Lys Leu Ser Phe Leu Arg Leu Ile Ser
1115 1120 1125
Asn Glu Ser Glu Gln Asp Arg Val Glu Phe Arg Lys Arg Ile Met
1130 1135 1140
Asn Thr Lys Lys Glu Asp Phe Tyr Lys Phe Ala Asp Leu Leu Glu
1145 1150 1155
Ser Lys Val Asn Glu Phe Glu Lys Asn Ile Val Ile Ile Thr Thr
1160 1165 1170
Lys Glu Lys Ala Asn Glu Tyr Ile Ala Asn Val Asp Gly Glu Phe
1175 1180 1185
Lys Lys Val Leu Ile Glu
1190
<210> 42
<211> 1193
<212> PRT
<213> 恶性疟原虫（Plasmodium falciparum）
<220>
<221> misc_feature
<223> Falcilysin
<400> 42
Met Asn Leu Thr Lys Leu Met Lys Val Ile Gly Tyr Ile Asn Ile Ile
1 5 10 15
Thr Asn Cys Val Gln Ser Phe Thr Asn Arg Ala Asp Lys Lys Arg Tyr
20 25 30
Asn Val Phe Ala Lys Ser Phe Ile Asn Thr Ile Asn Thr Asn Leu Tyr
35 40 45
Thr Phe Lys Ala Val Met Ser Lys Thr Pro Glu Trp Ile His Glu Lys
50 55 60
Ser Pro Lys His Asn Ser Tyr Asp Ile Ile Glu Lys Arg Tyr Asn Glu
65 70 75 80
Glu Phe Lys Met Thr Tyr Thr Val Tyr Gln His Lys Lys Ala Lys Thr
85 90 95
Gln Val Ile Ser Leu Gly Thr Asn Asp Pro Leu Asp Val Glu Gln Ala
100 105 110
Phe Ala Phe Tyr Val Lys Thr Leu Thr His Ser Gly Lys Gly Ile Pro
115 120 125
His Ile Leu Glu His Ser Val Leu Ser Gly Ser Lys Asn Tyr Asn Tyr
130 135 140
Lys Asn Ser Ile Gly Leu Leu Glu Lys Gly Thr Leu His Thr His Leu
145 150 155 160
Asn Ala Tyr Thr Phe Asn Asp Arg Thr Val Tyr Met Ala Gly Ser Met
165 170 175
Asn Asn Lys Asp Phe Phe Asn Ile Met Gly Val Tyr Met Asp Ser Val
180 185 190
Phe Gln Pro Asn Val Leu Glu Asn Lys Tyr Ile Phe Glu Thr Glu Gly
195 200 205
Trp Thr Tyr Glu Val Glu Lys Leu Lys Glu Asp Glu Lys Gly Lys Ala
210 215 220
Glu Ile Pro Gln Met Lys Asp Tyr Lys Val Ser Phe Asn Gly Ile Val
225 230 235 240
Tyr Asn Glu Met Lys Gly Ala Leu Ser Ser Pro Leu Glu Asp Leu Tyr
245 250 255
His Glu Glu Met Lys Tyr Met Phe Pro Asp Asn Val His Ser Asn Asn
260 265 270
Ser Gly Gly Asp Pro Lys Glu Ile Thr Asn Leu Thr Tyr Glu Glu Phe
275 280 285
Lys Glu Phe Tyr Tyr Lys Asn Tyr Asn Pro Lys Lys Val Lys Val Phe
290 295 300
Phe Phe Ser Lys Asn Asn Pro Thr Glu Leu Leu Asn Phe Val Asp Gln
305 310 315 320
Tyr Leu Gly Gln Leu Asp Tyr Ser Lys Tyr Arg Asp Asp Ala Val Glu
325 330 335
Ser Val Glu Tyr Gln Thr Tyr Lys Lys Gly Pro Phe Tyr Ile Lys Lys
340 345 350
Lys Tyr Gly Asp His Ser Glu Glu Lys Glu Asn Leu Val Ser Val Ala
355 360 365
Trp Leu Leu Asn Pro Lys Val Asp Lys Thr Asn Asn His Asn Asn Asn
370 375 380
His Ser Asn Asn Gln Ser Ser Glu Asn Asn Gly Tyr Ser Asn Gly Ser
385 390 395 400
His Ser Ser Asp Leu Ser Leu Glu Asn Pro Thr Asp Tyr Phe Val Leu
405 410 415
Leu Ile Ile Asn Asn Leu Leu Ile His Thr Pro Glu Ser Val Leu Tyr
420 425 430
Lys Ala Leu Thr Asp Cys Gly Leu Gly Asn Asn Val Ile Asp Arg Gly
435 440 445
Leu Asn Asp Ser Leu Val Gln Tyr Ile Phe Ser Ile Gly Leu Lys Gly
450 455 460
Ile Lys Arg Asn Asn Glu Lys Ile Lys Asn Phe Asp Lys Val His Tyr
465 470 475 480
Glu Val Glu Asp Val Ile Met Asn Ala Leu Lys Lys Val Val Lys Glu
485 490 495
Gly Phe Asn Lys Ser Ala Val Glu Ala Ser Ile Asn Asn Ile Glu Phe
500 505 510
Ile Leu Lys Glu Ala Asn Leu Lys Thr Ser Lys Ser Ile Asp Phe Val
515 520 525
Phe Glu Met Thr Ser Lys Leu Asn Tyr Asn Arg Asp Pro Leu Leu Ile
530 535 540
Phe Glu Phe Glu Lys Tyr Leu Asn Ile Val Lys Asn Lys Ile Lys Asn
545 550 555 560
Glu Pro Met Tyr Leu Glu Lys Phe Val Glu Lys His Phe Ile Asn Asn
565 570 575
Ala His Arg Ser Val Ile Leu Leu Glu Gly Asp Glu Asn Tyr Ala Gln
580 585 590
Glu Gln Glu Asn Leu Glu Lys Gln Glu Leu Lys Lys Arg Ile Glu Asn
595 600 605
Phe Asn Glu Gln Glu Lys Glu Gln Val Ile Lys Asn Phe Glu Glu Leu
610 615 620
Ser Lys Tyr Lys Asn Ala Glu Glu Ser Pro Glu His Leu Asn Lys Phe
625 630 635 640
Pro Ile Ile Ser Ile Ser Asp Leu Asn Lys Lys Thr Leu Glu Val Pro
645 650 655
Val Asn Val Tyr Phe Thr Asn Ile Asn Glu Asn Asn Asn Ile Met Glu
660 665 670
Thr Tyr Asn Lys Leu Lys Thr Asn Glu His Met Leu Lys Asp Asn Met
675 680 685
Asp Val Phe Leu Lys Lys Tyr Val Leu Lys Asn Asp Lys His Asn Thr
690 695 700
Asn Asn Asn Asn Asn Asn Asn Asn Asn Met Asp Tyr Ser Phe Thr Glu
705 710 715 720
Thr Lys Tyr Glu Gly Asn Val Pro Ile Leu Val Tyr Glu Met Pro Thr
725 730 735
Thr Gly Ile Val Tyr Leu Gln Phe Val Phe Ser Leu Asp His Leu Thr
740 745 750
Val Asp Glu Leu Ala Tyr Leu Asn Leu Phe Lys Thr Leu Ile Leu Glu
755 760 765
Asn Lys Thr Asn Lys Arg Ser Ser Glu Asp Phe Val Ile Leu Arg Glu
770 775 780
Lys Asn Ile Gly Ser Met Ser Ala Asn Val Ala Leu Tyr Ser Lys Asp
785 790 795 800
Asp His Leu Asn Val Thr Asp Lys Tyr Asn Ala Gln Ala Leu Phe Asn
805 810 815
Leu Glu Met His Val Leu Ser His Lys Cys Asn Asp Ala Leu Asn Ile
820 825 830
Ala Leu Glu Ala Val Lys Glu Ser Asp Phe Ser Asn Lys Lys Lys Val
835 840 845
Ile Asp Ile Leu Lys Arg Lys Ile Asn Gly Met Lys Thr Thr Phe Ser
850 855 860
Glu Lys Gly Tyr Ala Ile Leu Met Lys Tyr Val Lys Ala His Leu Asn
865 870 875 880
Ser Lys His Tyr Ala His Asn Ile Ile Tyr Gly Tyr Glu Asn Tyr Leu
885 890 895
Lys Leu Gln Glu Gln Leu Glu Leu Ala Glu Asn Asp Phe Lys Thr Leu
900 905 910
Glu Asn Ile Leu Val Arg Ile Arg Asn Lys Ile Phe Asn Lys Lys Asn
915 920 925
Leu Met Val Ser Val Thr Ser Asp Tyr Gly Ala Leu Lys His Leu Phe
930 935 940
Val Asn Ser Asn Glu Ser Leu Lys Asn Leu Val Ser Tyr Phe Glu Glu
945 950 955 960
Asn Asp Lys Tyr Ile Asn Asp Met Gln Asn Lys Val Asn Asp Pro Thr
965 970 975
Val Met Gly Trp Asn Glu Glu Ile Lys Ser Lys Lys Leu Phe Asp Glu
980 985 990
Glu Lys Val Lys Lys Glu Phe Phe Val Leu Pro Thr Phe Val Asn Ser
995 1000 1005
Val Ser Met Ser Gly Ile Leu Phe Lys Pro Gly Glu Tyr Leu Asp
1010 1015 1020
Pro Ser Phe Thr Val Ile Val Ala Ala Leu Lys Asn Ser Tyr Leu
1025 1030 1035
Trp Asp Thr Val Arg Gly Leu Asn Gly Ala Tyr Gly Val Phe Ala
1040 1045 1050
Asp Ile Glu Tyr Asp Gly Ser Val Val Phe Leu Ser Ala Arg Asp
1055 1060 1065
Pro Asn Leu Glu Lys Thr Leu Ala Thr Phe Arg Glu Ser Ala Lys
1070 1075 1080
Gly Leu Arg Lys Met Ala Asp Thr Met Thr Glu Asn Asp Leu Leu
1085 1090 1095
Arg Tyr Ile Ile Asn Thr Ile Gly Thr Ile Asp Lys Pro Arg Arg
1100 1105 1110
Gly Ile Glu Leu Ser Lys Leu Ser Phe Leu Arg Leu Ile Ser Asn
1115 1120 1125
Glu Ser Glu Gln Asp Arg Val Glu Phe Arg Lys Arg Ile Met Asn
1130 1135 1140
Thr Lys Lys Glu Asp Phe Tyr Lys Phe Ala Asp Leu Leu Glu Ser
1145 1150 1155
Lys Val Asn Glu Phe Glu Lys Asn Ile Val Ile Ile Thr Thr Lys
1160 1165 1170
Glu Lys Ala Asn Glu Tyr Ile Ala Asn Val Asp Gly Glu Phe Lys
1175 1180 1185
Lys Val Leu Ile Glu
1190
<210> 43
<211> 3471
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> PvFalcilysin 人 CO
<400> 43
gccaccatgg ctaaactgat gagggtcttt ggttatctta acataattac taactgcgtt 60
aacggacttc tctgtaaagc ggagaaacgc aaatataacg tctttacgaa cagctttatc 120
tattctatct ctacaacaaa tctgtactcc ttcacggcca tgatgaacaa atcacccgag 180
tgggtgcagg aaaagtgtcc cgagcacaag tcttataaca tccttgaaaa gcggttttct 240
gataaattcc aaatgacgta tacagtgtac gagcacaaaa aagcgaaaac ccaagtcatt 300
gccctcggca gcaatgaccc tctggacgtt gagcagactt ttgcctttta cgtaaagacc 360
ctgacgaact caggtaaggg tattcctcac atactcgagc acacagtttt gagcgggagc 420
aaaaatttca attacaaaga tagcatggga ctcctggaaa agggaacact taacacccat 480
ttgaatgcgt atactttcaa cgacaggact gtgtatatgg cggggtctat gaataataga 540
gactttttca acataatggc ggtgtatatg gatagcgtgt ttcaaccgaa tgttcttgaa 600
aataagttta tcttccaaac ggaaggatgg acctacgaag tggaaaagtt gaaagatgag 660
gagaagaacg ctgatgtacc gaaaattaaa gattataagg tttcctacaa tggtatagtc 720
tattccgaaa tgaaagggag cttctcctct cccctgcaat atctttatta tttgatcatg 780
aaaaacatct tccctgacaa cgtccattct aacataagtg gaggggaccc taaggagatt 840
ccaacgctca cgtatgagga attcaaagag ttctactata agaattacaa tcctaaaaaa 900
attaaagtca ttttcttcag caaaaacaat cctacagagt tgttgaattt cgtcgacgat 960
tatttgaacc agctcgactt tactaaatac agagacgatg cagtagaaaa cgttaattat 1020
caggagtaca agaaggggcc attctatgtt aagaagaaat ttgcggatca cagtgaggaa 1080
aaggaaaatc ttgtttccat ttcatggctt ctcaacccga aaaaaaatga cctcctggat 1140
gttgatctta gccttgaaag cccccaagac tacttcgctt tgttgattat taacaacctt 1200
ctgacgcaca ccaccgaatc tgtgctttac aaggcgctta ttgactgcgg tttgggtaac 1260
accgttatcg atacaggcct tgacgacagc ctggtacagt tcatattttc tataggtctg 1320
aagggtatca aggaaaagaa tgaaaagaac gtatcattgg acgttgtcca ttatgaagtg 1380
gaaaaggtgg ttctcaaagc gctccaaaaa gtagtggatg aaggcttcaa caaatcagcg 1440
gtcgaggcca gcattaacaa catagaattc gttttgaaag aagccaatct taaaacgtcc 1500
aaaagcgtgg actacatatt tgaaatggct tccaggctga attataatag agaccctctg 1560
ctgatctttg aattcgagaa gcacttgaac gtcgttaaag ataagatcaa gaatgaacca 1620
aagtatttgg agaaattcat tgagaaacac tttataaaca ataaccatcg ggctgtcatc 1680
ttgatggagg gtgatgaaaa ctatgggaaa gaacaggagg atttggaaaa ggagactttg 1740
aaaaagaaga tcgaatcact cacggaaaaa gaacgggacg acataatagt tgatttcgag 1800
aacctgacaa agtataagaa catggtcgag agccccgaac atttggataa ttttcccatc 1860
atctctatca gcgatctgaa caaagaaacg ctggaaatcc ctgctaatgc atatttcacg 1920
tccacggcag aggaaaataa tatggaaaag tacaataaag tgaaggcaag cgaggatgtt 1980
atgaagaaaa acatggacca cctcatcgac aaatatgttc tgaaaggagc gcaaggaggc 2040
gctgctaccg acggtgcagc caaacagggt gattccagcg atggtgaaat ccccatgctc 2100
gtttacgaga tgcctaccag tggaatcttg tatctgcaat ttatctttaa cctcgatcat 2160
ctgagcctgg aggaaatgtc atatctcaat ctctttaaga tgctcatcct tgagaacaaa 2220
actatgaaac gctcctctga ggagtttgtc attttgaggg aaaagaatat cggcaacata 2280
atggctaacg tcgccttgta tagtatcagc gatcacctca aggtcacttc taaatacaac 2340
gctcacggtc tgttcaattt tgaaatgcat gtattgagtc acaagtgcaa tgagtctttg 2400
gaaattgcct tggaagctct caaggactct gattttagca acaagaagaa aatagtggag 2460
atcctgaaac ggaagatcaa cggcatgaag acggtcttct cttctaaagg ctactcactt 2520
ctcctcaagt atgtaaagtc acaaatgaat gcaaaatact acgctcatga tttggttttt 2580
ggatacggca actatttgaa gttgcaagaa cagctcaagc tcgccgaaag tgactttcca 2640
cagttcgagc agattctcaa cagaatccga aataagattt tcactaagaa gaatctgctg 2700
atcagcgtga ccagtgacgc tgcagcgttg gatcaactgt ttgtgcatag caaggaatcc 2760
ctgaagaacc ttcttgggta tttcgaagag aatgatgcca agtctggaga ggctgagacc 2820
atagggtgga atgaggagat taaacaatca aaagtgatcg aaaaggaaca aaagaagaag 2880
gaattctttg taataccaac atttgtcaat gcagtatcaa tggctggaat gttgtttaat 2940
gagaaggagt tcctggatcc gtctttcata gttatcgtgg ccgcattgaa aaactcatac 3000
ctttgggaga ccgtgagggg actcaacggg gcatatggag tttttgctga tatagaatac 3060
gatggtgccg tcgtgttctt gtcagcccgc gatccgaatc tggaaaagac gctgcaaacg 3120
ttcaaggagt ctgctcaagg gttgagaaag atggcggata caatgaccaa aaatgacctt 3180
cggcgctaca taatcaatgc gatcggcaat attgataagc cgagacgggg tgttgagctc 3240
tcaaagctct cacttttgag aattatatct aatgaaacca aacaggaccg aatcgacttt 3300
cgcaagagga ttatggagac gacgaaggaa gacttctata aattcgctga cttgcttgaa 3360
aagaagattg cagaatttga aaagaatatt gttattatca catctaagga aaaagcctca 3420
gaatacagca ctaacgtaga ccaggatttt aagcagattc atatcgagta g 3471
<210> 44
<211> 1154
<212> PRT
<213> 间日疟原虫（Plasmodium vivax）
<220>
<221> misc_feature
<223> PvFalcilysin
<400> 44
Met Ala Lys Leu Met Arg Val Phe Gly Tyr Leu Asn Ile Ile Thr Asn
1 5 10 15
Cys Val Asn Gly Leu Leu Cys Lys Ala Glu Lys Arg Lys Tyr Asn Val
20 25 30
Phe Thr Asn Ser Phe Ile Tyr Ser Ile Ser Thr Thr Asn Leu Tyr Ser
35 40 45
Phe Thr Ala Met Met Asn Lys Ser Pro Glu Trp Val Gln Glu Lys Cys
50 55 60
Pro Glu His Lys Ser Tyr Asn Ile Leu Glu Lys Arg Phe Ser Asp Lys
65 70 75 80
Phe Gln Met Thr Tyr Thr Val Tyr Glu His Lys Lys Ala Lys Thr Gln
85 90 95
Val Ile Ala Leu Gly Ser Asn Asp Pro Leu Asp Val Glu Gln Thr Phe
100 105 110
Ala Phe Tyr Val Lys Thr Leu Thr Asn Ser Gly Lys Gly Ile Pro His
115 120 125
Ile Leu Glu His Thr Val Leu Ser Gly Ser Lys Asn Phe Asn Tyr Lys
130 135 140
Asp Ser Met Gly Leu Leu Glu Lys Gly Thr Leu Asn Thr His Leu Asn
145 150 155 160
Ala Tyr Thr Phe Asn Asp Arg Thr Val Tyr Met Ala Gly Ser Met Asn
165 170 175
Asn Arg Asp Phe Phe Asn Ile Met Ala Val Tyr Met Asp Ser Val Phe
180 185 190
Gln Pro Asn Val Leu Glu Asn Lys Phe Ile Phe Gln Thr Glu Gly Trp
195 200 205
Thr Tyr Glu Val Glu Lys Leu Lys Asp Glu Glu Lys Asn Ala Asp Val
210 215 220
Pro Lys Ile Lys Asp Tyr Lys Val Ser Tyr Asn Gly Ile Val Tyr Ser
225 230 235 240
Glu Met Lys Gly Ser Phe Ser Ser Pro Leu Gln Tyr Leu Tyr Tyr Leu
245 250 255
Ile Met Lys Asn Ile Phe Pro Asp Asn Val His Ser Asn Ile Ser Gly
260 265 270
Gly Asp Pro Lys Glu Ile Pro Thr Leu Thr Tyr Glu Glu Phe Lys Glu
275 280 285
Phe Tyr Tyr Lys Asn Tyr Asn Pro Lys Lys Ile Lys Val Ile Phe Phe
290 295 300
Ser Lys Asn Asn Pro Thr Glu Leu Leu Asn Phe Val Asp Asp Tyr Leu
305 310 315 320
Asn Gln Leu Asp Phe Thr Lys Tyr Arg Asp Asp Ala Val Glu Asn Val
325 330 335
Asn Tyr Gln Glu Tyr Lys Lys Gly Pro Phe Tyr Val Lys Lys Lys Phe
340 345 350
Ala Asp His Ser Glu Glu Lys Glu Asn Leu Val Ser Ile Ser Trp Leu
355 360 365
Leu Asn Pro Lys Lys Asn Asp Leu Leu Asp Val Asp Leu Ser Leu Glu
370 375 380
Ser Pro Gln Asp Tyr Phe Ala Leu Leu Ile Ile Asn Asn Leu Leu Thr
385 390 395 400
His Thr Thr Glu Ser Val Leu Tyr Lys Ala Leu Ile Asp Cys Gly Leu
405 410 415
Gly Asn Thr Val Ile Asp Thr Gly Leu Asp Asp Ser Leu Val Gln Phe
420 425 430
Ile Phe Ser Ile Gly Leu Lys Gly Ile Lys Glu Lys Asn Glu Lys Asn
435 440 445
Val Ser Leu Asp Val Val His Tyr Glu Val Glu Lys Val Val Leu Lys
450 455 460
Ala Leu Gln Lys Val Val Asp Glu Gly Phe Asn Lys Ser Ala Val Glu
465 470 475 480
Ala Ser Ile Asn Asn Ile Glu Phe Val Leu Lys Glu Ala Asn Leu Lys
485 490 495
Thr Ser Lys Ser Val Asp Tyr Ile Phe Glu Met Ala Ser Arg Leu Asn
500 505 510
Tyr Asn Arg Asp Pro Leu Leu Ile Phe Glu Phe Glu Lys His Leu Asn
515 520 525
Val Val Lys Asp Lys Ile Lys Asn Glu Pro Lys Tyr Leu Glu Lys Phe
530 535 540
Ile Glu Lys His Phe Ile Asn Asn Asn His Arg Ala Val Ile Leu Met
545 550 555 560
Glu Gly Asp Glu Asn Tyr Gly Lys Glu Gln Glu Asp Leu Glu Lys Glu
565 570 575
Thr Leu Lys Lys Lys Ile Glu Ser Leu Thr Glu Lys Glu Arg Asp Asp
580 585 590
Ile Ile Val Asp Phe Glu Asn Leu Thr Lys Tyr Lys Asn Met Val Glu
595 600 605
Ser Pro Glu His Leu Asp Asn Phe Pro Ile Ile Ser Ile Ser Asp Leu
610 615 620
Asn Lys Glu Thr Leu Glu Ile Pro Ala Asn Ala Tyr Phe Thr Ser Thr
625 630 635 640
Ala Glu Glu Asn Asn Met Glu Lys Tyr Asn Lys Val Lys Ala Ser Glu
645 650 655
Asp Val Met Lys Lys Asn Met Asp His Leu Ile Asp Lys Tyr Val Leu
660 665 670
Lys Gly Ala Gln Gly Gly Ala Ala Thr Asp Gly Ala Ala Lys Gln Gly
675 680 685
Asp Ser Ser Asp Gly Glu Ile Pro Met Leu Val Tyr Glu Met Pro Thr
690 695 700
Ser Gly Ile Leu Tyr Leu Gln Phe Ile Phe Asn Leu Asp His Leu Ser
705 710 715 720
Leu Glu Glu Met Ser Tyr Leu Asn Leu Phe Lys Met Leu Ile Leu Glu
725 730 735
Asn Lys Thr Met Lys Arg Ser Ser Glu Glu Phe Val Ile Leu Arg Glu
740 745 750
Lys Asn Ile Gly Asn Ile Met Ala Asn Val Ala Leu Tyr Ser Ile Ser
755 760 765
Asp His Leu Lys Val Thr Ser Lys Tyr Asn Ala His Gly Leu Phe Asn
770 775 780
Phe Glu Met His Val Leu Ser His Lys Cys Asn Glu Ser Leu Glu Ile
785 790 795 800
Ala Leu Glu Ala Leu Lys Asp Ser Asp Phe Ser Asn Lys Lys Lys Ile
805 810 815
Val Glu Ile Leu Lys Arg Lys Ile Asn Gly Met Lys Thr Val Phe Ser
820 825 830
Ser Lys Gly Tyr Ser Leu Leu Leu Lys Tyr Val Lys Ser Gln Met Asn
835 840 845
Ala Lys Tyr Tyr Ala His Asp Leu Val Phe Gly Tyr Gly Asn Tyr Leu
850 855 860
Lys Leu Gln Glu Gln Leu Lys Leu Ala Glu Ser Asp Phe Pro Gln Phe
865 870 875 880
Glu Gln Ile Leu Asn Arg Ile Arg Asn Lys Ile Phe Thr Lys Lys Asn
885 890 895
Leu Leu Ile Ser Val Thr Ser Asp Ala Ala Ala Leu Asp Gln Leu Phe
900 905 910
Val His Ser Lys Glu Ser Leu Lys Asn Leu Leu Gly Tyr Phe Glu Glu
915 920 925
Asn Asp Ala Lys Ser Gly Glu Ala Glu Thr Ile Gly Trp Asn Glu Glu
930 935 940
Ile Lys Gln Ser Lys Val Ile Glu Lys Glu Gln Lys Lys Lys Glu Phe
945 950 955 960
Phe Val Ile Pro Thr Phe Val Asn Ala Val Ser Met Ala Gly Met Leu
965 970 975
Phe Asn Glu Lys Glu Phe Leu Asp Pro Ser Phe Ile Val Ile Val Ala
980 985 990
Ala Leu Lys Asn Ser Tyr Leu Trp Glu Thr Val Arg Gly Leu Asn Gly
995 1000 1005
Ala Tyr Gly Val Phe Ala Asp Ile Glu Tyr Asp Gly Ala Val Val
1010 1015 1020
Phe Leu Ser Ala Arg Asp Pro Asn Leu Glu Lys Thr Leu Gln Thr
1025 1030 1035
Phe Lys Glu Ser Ala Gln Gly Leu Arg Lys Met Ala Asp Thr Met
1040 1045 1050
Thr Lys Asn Asp Leu Arg Arg Tyr Ile Ile Asn Ala Ile Gly Asn
1055 1060 1065
Ile Asp Lys Pro Arg Arg Gly Val Glu Leu Ser Lys Leu Ser Leu
1070 1075 1080
Leu Arg Ile Ile Ser Asn Glu Thr Lys Gln Asp Arg Ile Asp Phe
1085 1090 1095
Arg Lys Arg Ile Met Glu Thr Thr Lys Glu Asp Phe Tyr Lys Phe
1100 1105 1110
Ala Asp Leu Leu Glu Lys Lys Ile Ala Glu Phe Glu Lys Asn Ile
1115 1120 1125
Val Ile Ile Thr Ser Lys Glu Lys Ala Ser Glu Tyr Ser Thr Asn
1130 1135 1140
Val Asp Gln Asp Phe Lys Gln Ile His Ile Glu
1145 1150
<210> 45
<211> 1153
<212> PRT
<213> 间日疟原虫（Plasmodium vivax）
<220>
<221> misc_feature
<223> Falcilysin
<400> 45
Met Lys Leu Met Arg Val Phe Gly Tyr Leu Asn Ile Ile Thr Asn Cys
1 5 10 15
Val Asn Gly Leu Leu Cys Lys Ala Glu Lys Arg Lys Tyr Asn Val Phe
20 25 30
Thr Asn Ser Phe Ile Tyr Ser Ile Ser Thr Thr Asn Leu Tyr Ser Phe
35 40 45
Thr Ala Met Met Asn Lys Ser Pro Glu Trp Val Gln Glu Lys Cys Pro
50 55 60
Glu His Lys Ser Tyr Asn Ile Leu Glu Lys Arg Phe Ser Asp Lys Phe
65 70 75 80
Gln Met Thr Tyr Thr Val Tyr Glu His Lys Lys Ala Lys Thr Gln Val
85 90 95
Ile Ala Leu Gly Ser Asn Asp Pro Leu Asp Val Glu Gln Thr Phe Ala
100 105 110
Phe Tyr Val Lys Thr Leu Thr Asn Ser Gly Lys Gly Ile Pro His Ile
115 120 125
Leu Glu His Thr Val Leu Ser Gly Ser Lys Asn Phe Asn Tyr Lys Asp
130 135 140
Ser Met Gly Leu Leu Glu Lys Gly Thr Leu Asn Thr His Leu Asn Ala
145 150 155 160
Tyr Thr Phe Asn Asp Arg Thr Val Tyr Met Ala Gly Ser Met Asn Asn
165 170 175
Arg Asp Phe Phe Asn Ile Met Ala Val Tyr Met Asp Ser Val Phe Gln
180 185 190
Pro Asn Val Leu Glu Asn Lys Phe Ile Phe Gln Thr Glu Gly Trp Thr
195 200 205
Tyr Glu Val Glu Lys Leu Lys Asp Glu Glu Lys Asn Ala Asp Val Pro
210 215 220
Lys Ile Lys Asp Tyr Lys Val Ser Tyr Asn Gly Ile Val Tyr Ser Glu
225 230 235 240
Met Lys Gly Ser Phe Ser Ser Pro Leu Gln Tyr Leu Tyr Tyr Leu Ile
245 250 255
Met Lys Asn Ile Phe Pro Asp Asn Val His Ser Asn Ile Ser Gly Gly
260 265 270
Asp Pro Lys Glu Ile Pro Thr Leu Thr Tyr Glu Glu Phe Lys Glu Phe
275 280 285
Tyr Tyr Lys Asn Tyr Asn Pro Lys Lys Ile Lys Val Ile Phe Phe Ser
290 295 300
Lys Asn Asn Pro Thr Glu Leu Leu Asn Phe Val Asp Asp Tyr Leu Asn
305 310 315 320
Gln Leu Asp Phe Thr Lys Tyr Arg Asp Asp Ala Val Glu Asn Val Asn
325 330 335
Tyr Gln Glu Tyr Lys Lys Gly Pro Phe Tyr Val Lys Lys Lys Phe Ala
340 345 350
Asp His Ser Glu Glu Lys Glu Asn Leu Val Ser Ile Ser Trp Leu Leu
355 360 365
Asn Pro Lys Lys Asn Asp Leu Leu Asp Val Asp Leu Ser Leu Glu Ser
370 375 380
Pro Gln Asp Tyr Phe Ala Leu Leu Ile Ile Asn Asn Leu Leu Thr His
385 390 395 400
Thr Thr Glu Ser Val Leu Tyr Lys Ala Leu Ile Asp Cys Gly Leu Gly
405 410 415
Asn Thr Val Ile Asp Thr Gly Leu Asp Asp Ser Leu Val Gln Phe Ile
420 425 430
Phe Ser Ile Gly Leu Lys Gly Ile Lys Glu Lys Asn Glu Lys Asn Val
435 440 445
Ser Leu Asp Val Val His Tyr Glu Val Glu Lys Val Val Leu Lys Ala
450 455 460
Leu Gln Lys Val Val Asp Glu Gly Phe Asn Lys Ser Ala Val Glu Ala
465 470 475 480
Ser Ile Asn Asn Ile Glu Phe Val Leu Lys Glu Ala Asn Leu Lys Thr
485 490 495
Ser Lys Ser Val Asp Tyr Ile Phe Glu Met Ala Ser Arg Leu Asn Tyr
500 505 510
Asn Arg Asp Pro Leu Leu Ile Phe Glu Phe Glu Lys His Leu Asn Val
515 520 525
Val Lys Asp Lys Ile Lys Asn Glu Pro Lys Tyr Leu Glu Lys Phe Ile
530 535 540
Glu Lys His Phe Ile Asn Asn Asn His Arg Ala Val Ile Leu Met Glu
545 550 555 560
Gly Asp Glu Asn Tyr Gly Lys Glu Gln Glu Asp Leu Glu Lys Glu Thr
565 570 575
Leu Lys Lys Lys Ile Glu Ser Leu Thr Glu Lys Glu Arg Asp Asp Ile
580 585 590
Ile Val Asp Phe Glu Asn Leu Thr Lys Tyr Lys Asn Met Val Glu Ser
595 600 605
Pro Glu His Leu Asp Asn Phe Pro Ile Ile Ser Ile Ser Asp Leu Asn
610 615 620
Lys Glu Thr Leu Glu Ile Pro Ala Asn Ala Tyr Phe Thr Ser Thr Ala
625 630 635 640
Glu Glu Asn Asn Met Glu Lys Tyr Asn Lys Val Lys Ala Ser Glu Asp
645 650 655
Val Met Lys Lys Asn Met Asp His Leu Ile Asp Lys Tyr Val Leu Lys
660 665 670
Gly Ala Gln Gly Gly Ala Ala Thr Asp Gly Ala Ala Lys Gln Gly Asp
675 680 685
Ser Ser Asp Gly Glu Ile Pro Met Leu Val Tyr Glu Met Pro Thr Ser
690 695 700
Gly Ile Leu Tyr Leu Gln Phe Ile Phe Asn Leu Asp His Leu Ser Leu
705 710 715 720
Glu Glu Met Ser Tyr Leu Asn Leu Phe Lys Met Leu Ile Leu Glu Asn
725 730 735
Lys Thr Met Lys Arg Ser Ser Glu Glu Phe Val Ile Leu Arg Glu Lys
740 745 750
Asn Ile Gly Asn Ile Met Ala Asn Val Ala Leu Tyr Ser Ile Ser Asp
755 760 765
His Leu Lys Val Thr Ser Lys Tyr Asn Ala His Gly Leu Phe Asn Phe
770 775 780
Glu Met His Val Leu Ser His Lys Cys Asn Glu Ser Leu Glu Ile Ala
785 790 795 800
Leu Glu Ala Leu Lys Asp Ser Asp Phe Ser Asn Lys Lys Lys Ile Val
805 810 815
Glu Ile Leu Lys Arg Lys Ile Asn Gly Met Lys Thr Val Phe Ser Ser
820 825 830
Lys Gly Tyr Ser Leu Leu Leu Lys Tyr Val Lys Ser Gln Met Asn Ala
835 840 845
Lys Tyr Tyr Ala His Asp Leu Val Phe Gly Tyr Gly Asn Tyr Leu Lys
850 855 860
Leu Gln Glu Gln Leu Lys Leu Ala Glu Ser Asp Phe Pro Gln Phe Glu
865 870 875 880
Gln Ile Leu Asn Arg Ile Arg Asn Lys Ile Phe Thr Lys Lys Asn Leu
885 890 895
Leu Ile Ser Val Thr Ser Asp Ala Ala Ala Leu Asp Gln Leu Phe Val
900 905 910
His Ser Lys Glu Ser Leu Lys Asn Leu Leu Gly Tyr Phe Glu Glu Asn
915 920 925
Asp Ala Lys Ser Gly Glu Ala Glu Thr Ile Gly Trp Asn Glu Glu Ile
930 935 940
Lys Gln Ser Lys Val Ile Glu Lys Glu Gln Lys Lys Lys Glu Phe Phe
945 950 955 960
Val Ile Pro Thr Phe Val Asn Ala Val Ser Met Ala Gly Met Leu Phe
965 970 975
Asn Glu Lys Glu Phe Leu Asp Pro Ser Phe Ile Val Ile Val Ala Ala
980 985 990
Leu Lys Asn Ser Tyr Leu Trp Glu Thr Val Arg Gly Leu Asn Gly Ala
995 1000 1005
Tyr Gly Val Phe Ala Asp Ile Glu Tyr Asp Gly Ala Val Val Phe
1010 1015 1020
Leu Ser Ala Arg Asp Pro Asn Leu Glu Lys Thr Leu Gln Thr Phe
1025 1030 1035
Lys Glu Ser Ala Gln Gly Leu Arg Lys Met Ala Asp Thr Met Thr
1040 1045 1050
Lys Asn Asp Leu Arg Arg Tyr Ile Ile Asn Ala Ile Gly Asn Ile
1055 1060 1065
Asp Lys Pro Arg Arg Gly Val Glu Leu Ser Lys Leu Ser Leu Leu
1070 1075 1080
Arg Ile Ile Ser Asn Glu Thr Lys Gln Asp Arg Ile Asp Phe Arg
1085 1090 1095
Lys Arg Ile Met Glu Thr Thr Lys Glu Asp Phe Tyr Lys Phe Ala
1100 1105 1110
Asp Leu Leu Glu Lys Lys Ile Ala Glu Phe Glu Lys Asn Ile Val
1115 1120 1125
Ile Ile Thr Ser Lys Glu Lys Ala Ser Glu Tyr Ser Thr Asn Val
1130 1135 1140
Asp Gln Asp Phe Lys Gln Ile His Ile Glu
1145 1150
<210> 46
<211> 1227
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> Falcilysin，Bergheilysin-B-CO
<400> 46
ggatccgcca ccatggctaa actcatgaag gtcttggggt atatcaacat catcacgaat 60
tgcgtgaatg gcatactgtg taaaggcgat aagaagcgat acatgcacat gctctcccat 120
aagtgtaacg acgctctgga gattgcgctg gaagccctga aagaaagcga tttcagcaat 180
aagaagaaag tgattgaaat cctgaagagg aaaatcaatg gcatgaaaac cacgtttgcc 240
agcaaaggtc actcaatact gattaagtac gtgaagtctc ggattaatag caaatattac 300
gcttatgacc tgattcatgg gtatgacaac tatctgaagc tccaggaaca gcttaaactg 360
gcggagacaa actacgagag tctggaagcc atacttaatc gcatcaggaa gaagatcttt 420
aagcggaaca atctgatcat gaacgtgact gtcgatccag gaactattga tcagctgttt 480
gccaaatcca agaatagctt caacaacctt ctgtcatact ttgacgagaa cgagagctac 540
tgctctaaga atgactcctt caacaaagtg gtaggctgga acaaggagat tcaggagaag 600
aagctgcttg aaggagagga ggtcaagaaa gagctcttgg tcgtacccac atttgtgaac 660
agtgtctcta tgagtggagt gctgttcaac aaaggcgaat atcttgaccc ctcattcacc 720
gtaatcgttg cagccttgaa aaactcatac ctttgggaga cagttcgagg actgaatgga 780
gcttatgggg tgtttgccga catagagtat gacggtactg tcgttttcct gagtgcaaga 840
gatccgaatc tggaaaagac tctccaaacc tttcgtgaag cagcacaagg tctgcgtaaa 900
atggctgatg tgatgacaaa gaatgatctt ctgcggtata tcattaacgc tattgggacc 960
atcgatagac ctcgcagagg cgttgagttg tccaagctgt ccttctctcg cataataagc 1020
aacgaaaccg aacaggacag gattgaattc aggaaccggg tgatgaatac caagaaagag 1080
gatttctaca aatttgcgga tttgctcgaa aagaaagtca aggaatttga gaagaatgtg 1140
gtgatcatca catctaagga gaaagccaac gagtacatca acaatgttga caatgacttc 1200
aagaagatcc tcattgagta actcgag 1227
<210> 47
<211> 402
<212> PRT
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> Falcilysin，Bergheilysin-B
<400> 47
Met Ala Lys Leu Met Lys Val Leu Gly Tyr Ile Asn Ile Ile Thr Asn
1 5 10 15
Cys Val Asn Gly Ile Leu Cys Lys Gly Asp Lys Lys Arg Tyr Met His
20 25 30
Met Leu Ser His Lys Cys Asn Asp Ala Leu Glu Ile Ala Leu Glu Ala
35 40 45
Leu Lys Glu Ser Asp Phe Ser Asn Lys Lys Lys Val Ile Glu Ile Leu
50 55 60
Lys Arg Lys Ile Asn Gly Met Lys Thr Thr Phe Ala Ser Lys Gly His
65 70 75 80
Ser Ile Leu Ile Lys Tyr Val Lys Ser Arg Ile Asn Ser Lys Tyr Tyr
85 90 95
Ala Tyr Asp Leu Ile His Gly Tyr Asp Asn Tyr Leu Lys Leu Gln Glu
100 105 110
Gln Leu Lys Leu Ala Glu Thr Asn Tyr Glu Ser Leu Glu Ala Ile Leu
115 120 125
Asn Arg Ile Arg Lys Lys Ile Phe Lys Arg Asn Asn Leu Ile Met Asn
130 135 140
Val Thr Val Asp Pro Gly Thr Ile Asp Gln Leu Phe Ala Lys Ser Lys
145 150 155 160
Asn Ser Phe Asn Asn Leu Leu Ser Tyr Phe Asp Glu Asn Glu Ser Tyr
165 170 175
Cys Ser Lys Asn Asp Ser Phe Asn Lys Val Val Gly Trp Asn Lys Glu
180 185 190
Ile Gln Glu Lys Lys Leu Leu Glu Gly Glu Glu Val Lys Lys Glu Leu
195 200 205
Leu Val Val Pro Thr Phe Val Asn Ser Val Ser Met Ser Gly Val Leu
210 215 220
Phe Asn Lys Gly Glu Tyr Leu Asp Pro Ser Phe Thr Val Ile Val Ala
225 230 235 240
Ala Leu Lys Asn Ser Tyr Leu Trp Glu Thr Val Arg Gly Leu Asn Gly
245 250 255
Ala Tyr Gly Val Phe Ala Asp Ile Glu Tyr Asp Gly Thr Val Val Phe
260 265 270
Leu Ser Ala Arg Asp Pro Asn Leu Glu Lys Thr Leu Gln Thr Phe Arg
275 280 285
Glu Ala Ala Gln Gly Leu Arg Lys Met Ala Asp Val Met Thr Lys Asn
290 295 300
Asp Leu Leu Arg Tyr Ile Ile Asn Ala Ile Gly Thr Ile Asp Arg Pro
305 310 315 320
Arg Arg Gly Val Glu Leu Ser Lys Leu Ser Phe Ser Arg Ile Ile Ser
325 330 335
Asn Glu Thr Glu Gln Asp Arg Ile Glu Phe Arg Asn Arg Val Met Asn
340 345 350
Thr Lys Lys Glu Asp Phe Tyr Lys Phe Ala Asp Leu Leu Glu Lys Lys
355 360 365
Val Lys Glu Phe Glu Lys Asn Val Val Ile Ile Thr Ser Lys Glu Lys
370 375 380
Ala Asn Glu Tyr Ile Asn Asn Val Asp Asn Asp Phe Lys Lys Ile Leu
385 390 395 400
Ile Glu
<210> 48
<211> 2454
<212> DNA
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> 穿孔素样蛋白1（SPECT2）
<400> 48
ggatccgcca ccatggctaa gatgcggaac attaagaaat cacttccggt gctcttcatt 60
ctcttgtgca tttatcagca gtactttata aactctctcc gtatttccgt ccgcaacaat 120
aagaaccaca gggatgaaaa caaattcaac aagaatatgg agcttggtac gatggagaaa 180
cccatcaaca ttctttgcaa tgacgtcagc tgtaacacag agaataacat ttctttcgta 240
aaccagaaga aaaaggaaat agatagtgac agcgacctct ataacatgtt cgatgacgac 300
gcctctacct ccgctggtga tgacgaagat gactatgacg actacacaga tgataagaac 360
gctgagatca aagatgaaga gcaaaatgag cacatcgaca agatcgacca gaagaaggat 420
aagaaacgca cattctctat caataagcag gaggaagaaa tcaacgaaaa taaaaacaaa 480
acggagaaat tcttcaagaa atacaagttt aacgacgcca actcagaagg ggacgacgat 540
gagtcagaca ccgatgacga gaatttggac aactccacgg agaacagcta cgaggaaaac 600
aagaatcccg agaacgttat cgacaagcat atggccgtat ttcctgggct ctattttgtg 660
gggatcggct acgatattct gttcgggaat cctctcggag aaaccgattc tctgagcgac 720
cctggttata gagcacagat ttacctgttg aattgggaat tctcaaacca tggcatcgcc 780
aacgatctgc acacgctcca accaattaat gcctggattc ggaaagagaa tgcatgctca 840
cgagttgaat ccatcaacga atgttcctct gttagtgagt atacaaagaa tctgtcagtg 900
gacgtaagtg tatcaggcag ctatatgggc ttcggcagtt tctccgctag cactgggtac 960
aagaaattca ttaacgagat aagcaagaga acatccaaga cctactttat aaagagcaac 1020
tgcatcaagt ataccatcgg acttccacct tatgtgccat gggagcatac cactgcttac 1080
atgaatgcgg tgaatattct gccaaaggaa tttaccggcc tggatggaga cagcgaatgc 1140
acaccggatg tttacgagca gaagaagatg actaaacagt gtaagaacgt gcaactgtgg 1200
attcagttct ttaagaccta tggtacacac ataatcgtgg aggctcaatt gggagggaaa 1260
ataaccaaaa tcatcaatgt ctctaacaca agcgtgaacc agatgaagaa ggatggcgtc 1320
agtgtgaaag cccagatcca ggctcagttt ggttttgcaa gtgtgggcgg tagtacaagc 1380
gtgtcaagtg acaatagcac taagaacgac aatagcagct acgacatgtc tgagaaactc 1440
gtggttatcg gagggaatcc tataaaagac gtcaccaaag aggagaatct gtacgaatgg 1500
agcaaaacag tgtcctctaa ccccatgcct atccacatca agctgctgcc aatctataag 1560
tccttcgata gtgaggaact gaaagagtct tacgagaaag cggttctcta ctacaccagg 1620
ctttatggca gctctcccca cggaactatt cagaaagatg agaacgacat catcaaaatc 1680
ttgacggcca gtaccaccat cactaaaatt ggtgctccac ccataacagc ggaatgtcca 1740
cataatcaag tggtgctgtt tgggtatgtc ctgaagcaga acttctggga caacacctcc 1800
aacctgaagg gatacgacat tgagatctgt gaggctggac tgaatagttg cacgtccaaa 1860
cagggaagta caaacaagta cgatgtgagc tatctgtaca ttgaatgtgg cacacaggca 1920
atgtcattct ccgatcaagt cataaccgca tccaacacta cttacaatac catcaagtgt 1980
cccaatgact acactattat ctttggattc gggttttcct ctagctccgg taagggagtt 2040
tctgccatgc acacccacat tacatcctgt agacccggca tgaaaagctg ctccctgaat 2100
atgggcaaca gcaatgacaa gaactacatg tacctggtgt gcgtcgatgc cacaatctgg 2160
tctggcatta atgagctgac tattgtggcc aaagatgatt ttcacggcgc agtgaatagg 2220
tctaagcagt tcaatgatgg cgaattggta ctgtcctgtc aggaaaatgg cactatcctg 2280
acagggttca ctggggagac tcatacctct agcccgtatg tcaagagccc ttttagcaag 2340
tgtcttaaat cactgaagag ctgctcagtc catgggagtg gacaatctat cggatatacc 2400
aactataagt cactgttttc catcatactg tgcaagaacg gtgagtgact cgag 2454
<210> 49
<211> 811
<212> PRT
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> 穿孔素样蛋白1（SPECT2）
<400> 49
Met Ala Lys Met Arg Asn Ile Lys Lys Ser Leu Pro Val Leu Phe Ile
1 5 10 15
Leu Leu Cys Ile Tyr Gln Gln Tyr Phe Ile Asn Ser Leu Arg Ile Ser
20 25 30
Val Arg Asn Asn Lys Asn His Arg Asp Glu Asn Lys Phe Asn Lys Asn
35 40 45
Met Glu Leu Gly Thr Met Glu Lys Pro Ile Asn Ile Leu Cys Asn Asp
50 55 60
Val Ser Cys Asn Thr Glu Asn Asn Ile Ser Phe Val Asn Gln Lys Lys
65 70 75 80
Lys Glu Ile Asp Ser Asp Ser Asp Leu Tyr Asn Met Phe Asp Asp Asp
85 90 95
Ala Ser Thr Ser Ala Gly Asp Asp Glu Asp Asp Tyr Asp Asp Tyr Thr
100 105 110
Asp Asp Lys Asn Ala Glu Ile Lys Asp Glu Glu Gln Asn Glu His Ile
115 120 125
Asp Lys Ile Asp Gln Lys Lys Asp Lys Lys Arg Thr Phe Ser Ile Asn
130 135 140
Lys Gln Glu Glu Glu Ile Asn Glu Asn Lys Asn Lys Thr Glu Lys Phe
145 150 155 160
Phe Lys Lys Tyr Lys Phe Asn Asp Ala Asn Ser Glu Gly Asp Asp Asp
165 170 175
Glu Ser Asp Thr Asp Asp Glu Asn Leu Asp Asn Ser Thr Glu Asn Ser
180 185 190
Tyr Glu Glu Asn Lys Asn Pro Glu Asn Val Ile Asp Lys His Met Ala
195 200 205
Val Phe Pro Gly Leu Tyr Phe Val Gly Ile Gly Tyr Asp Ile Leu Phe
210 215 220
Gly Asn Pro Leu Gly Glu Thr Asp Ser Leu Ser Asp Pro Gly Tyr Arg
225 230 235 240
Ala Gln Ile Tyr Leu Leu Asn Trp Glu Phe Ser Asn His Gly Ile Ala
245 250 255
Asn Asp Leu His Thr Leu Gln Pro Ile Asn Ala Trp Ile Arg Lys Glu
260 265 270
Asn Ala Cys Ser Arg Val Glu Ser Ile Asn Glu Cys Ser Ser Val Ser
275 280 285
Glu Tyr Thr Lys Asn Leu Ser Val Asp Val Ser Val Ser Gly Ser Tyr
290 295 300
Met Gly Phe Gly Ser Phe Ser Ala Ser Thr Gly Tyr Lys Lys Phe Ile
305 310 315 320
Asn Glu Ile Ser Lys Arg Thr Ser Lys Thr Tyr Phe Ile Lys Ser Asn
325 330 335
Cys Ile Lys Tyr Thr Ile Gly Leu Pro Pro Tyr Val Pro Trp Glu His
340 345 350
Thr Thr Ala Tyr Met Asn Ala Val Asn Ile Leu Pro Lys Glu Phe Thr
355 360 365
Gly Leu Asp Gly Asp Ser Glu Cys Thr Pro Asp Val Tyr Glu Gln Lys
370 375 380
Lys Met Thr Lys Gln Cys Lys Asn Val Gln Leu Trp Ile Gln Phe Phe
385 390 395 400
Lys Thr Tyr Gly Thr His Ile Ile Val Glu Ala Gln Leu Gly Gly Lys
405 410 415
Ile Thr Lys Ile Ile Asn Val Ser Asn Thr Ser Val Asn Gln Met Lys
420 425 430
Lys Asp Gly Val Ser Val Lys Ala Gln Ile Gln Ala Gln Phe Gly Phe
435 440 445
Ala Ser Val Gly Gly Ser Thr Ser Val Ser Ser Asp Asn Ser Thr Lys
450 455 460
Asn Asp Asn Ser Ser Tyr Asp Met Ser Glu Lys Leu Val Val Ile Gly
465 470 475 480
Gly Asn Pro Ile Lys Asp Val Thr Lys Glu Glu Asn Leu Tyr Glu Trp
485 490 495
Ser Lys Thr Val Ser Ser Asn Pro Met Pro Ile His Ile Lys Leu Leu
500 505 510
Pro Ile Tyr Lys Ser Phe Asp Ser Glu Glu Leu Lys Glu Ser Tyr Glu
515 520 525
Lys Ala Val Leu Tyr Tyr Thr Arg Leu Tyr Gly Ser Ser Pro His Gly
530 535 540
Thr Ile Gln Lys Asp Glu Asn Asp Ile Ile Lys Ile Leu Thr Ala Ser
545 550 555 560
Thr Thr Ile Thr Lys Ile Gly Ala Pro Pro Ile Thr Ala Glu Cys Pro
565 570 575
His Asn Gln Val Val Leu Phe Gly Tyr Val Leu Lys Gln Asn Phe Trp
580 585 590
Asp Asn Thr Ser Asn Leu Lys Gly Tyr Asp Ile Glu Ile Cys Glu Ala
595 600 605
Gly Leu Asn Ser Cys Thr Ser Lys Gln Gly Ser Thr Asn Lys Tyr Asp
610 615 620
Val Ser Tyr Leu Tyr Ile Glu Cys Gly Thr Gln Ala Met Ser Phe Ser
625 630 635 640
Asp Gln Val Ile Thr Ala Ser Asn Thr Thr Tyr Asn Thr Ile Lys Cys
645 650 655
Pro Asn Asp Tyr Thr Ile Ile Phe Gly Phe Gly Phe Ser Ser Ser Ser
660 665 670
Gly Lys Gly Val Ser Ala Met His Thr His Ile Thr Ser Cys Arg Pro
675 680 685
Gly Met Lys Ser Cys Ser Leu Asn Met Gly Asn Ser Asn Asp Lys Asn
690 695 700
Tyr Met Tyr Leu Val Cys Val Asp Ala Thr Ile Trp Ser Gly Ile Asn
705 710 715 720
Glu Leu Thr Ile Val Ala Lys Asp Asp Phe His Gly Ala Val Asn Arg
725 730 735
Ser Lys Gln Phe Asn Asp Gly Glu Leu Val Leu Ser Cys Gln Glu Asn
740 745 750
Gly Thr Ile Leu Thr Gly Phe Thr Gly Glu Thr His Thr Ser Ser Pro
755 760 765
Tyr Val Lys Ser Pro Phe Ser Lys Cys Leu Lys Ser Leu Lys Ser Cys
770 775 780
Ser Val His Gly Ser Gly Gln Ser Ile Gly Tyr Thr Asn Tyr Lys Ser
785 790 795 800
Leu Phe Ser Ile Ile Leu Cys Lys Asn Gly Glu
805 810
<210> 50
<211> 810
<212> PRT
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> SPECT2
<400> 50
Met Lys Met Arg Asn Ile Lys Lys Ser Leu Pro Val Leu Phe Ile Leu
1 5 10 15
Leu Cys Ile Tyr Gln Gln Tyr Phe Ile Asn Ser Leu Arg Ile Ser Val
20 25 30
Arg Asn Asn Lys Asn His Arg Asp Glu Asn Lys Phe Asn Lys Asn Met
35 40 45
Glu Leu Gly Thr Met Glu Lys Pro Ile Asn Ile Leu Cys Asn Asp Val
50 55 60
Ser Cys Asn Thr Glu Asn Asn Ile Ser Phe Val Asn Gln Lys Lys Lys
65 70 75 80
Glu Ile Asp Ser Asp Ser Asp Leu Tyr Asn Met Phe Asp Asp Asp Ala
85 90 95
Ser Thr Ser Ala Gly Asp Asp Glu Asp Asp Tyr Asp Asp Tyr Thr Asp
100 105 110
Asp Lys Asn Ala Glu Ile Lys Asp Glu Glu Gln Asn Glu His Ile Asp
115 120 125
Lys Ile Asp Gln Lys Lys Asp Lys Lys Arg Thr Phe Ser Ile Asn Lys
130 135 140
Gln Glu Glu Glu Ile Asn Glu Asn Lys Asn Lys Thr Glu Lys Phe Phe
145 150 155 160
Lys Lys Tyr Lys Phe Asn Asp Ala Asn Ser Glu Gly Asp Asp Asp Glu
165 170 175
Ser Asp Thr Asp Asp Glu Asn Leu Asp Asn Ser Thr Glu Asn Ser Tyr
180 185 190
Glu Glu Asn Lys Asn Pro Glu Asn Val Ile Asp Lys His Met Ala Val
195 200 205
Phe Pro Gly Leu Tyr Phe Val Gly Ile Gly Tyr Asp Ile Leu Phe Gly
210 215 220
Asn Pro Leu Gly Glu Thr Asp Ser Leu Ser Asp Pro Gly Tyr Arg Ala
225 230 235 240
Gln Ile Tyr Leu Leu Asn Trp Glu Phe Ser Asn His Gly Ile Ala Asn
245 250 255
Asp Leu His Thr Leu Gln Pro Ile Asn Ala Trp Ile Arg Lys Glu Asn
260 265 270
Ala Cys Ser Arg Val Glu Ser Ile Asn Glu Cys Ser Ser Val Ser Glu
275 280 285
Tyr Thr Lys Asn Leu Ser Val Asp Val Ser Val Ser Gly Ser Tyr Met
290 295 300
Gly Phe Gly Ser Phe Ser Ala Ser Thr Gly Tyr Lys Lys Phe Ile Asn
305 310 315 320
Glu Ile Ser Lys Arg Thr Ser Lys Thr Tyr Phe Ile Lys Ser Asn Cys
325 330 335
Ile Lys Tyr Thr Ile Gly Leu Pro Pro Tyr Val Pro Trp Glu His Thr
340 345 350
Thr Ala Tyr Met Asn Ala Val Asn Ile Leu Pro Lys Glu Phe Thr Gly
355 360 365
Leu Asp Gly Asp Ser Glu Cys Thr Pro Asp Val Tyr Glu Gln Lys Lys
370 375 380
Met Thr Lys Gln Cys Lys Asn Val Gln Leu Trp Ile Gln Phe Phe Lys
385 390 395 400
Thr Tyr Gly Thr His Ile Ile Val Glu Ala Gln Leu Gly Gly Lys Ile
405 410 415
Thr Lys Ile Ile Asn Val Ser Asn Thr Ser Val Asn Gln Met Lys Lys
420 425 430
Asp Gly Val Ser Val Lys Ala Gln Ile Gln Ala Gln Phe Gly Phe Ala
435 440 445
Ser Val Gly Gly Ser Thr Ser Val Ser Ser Asp Asn Ser Thr Lys Asn
450 455 460
Asp Asn Ser Ser Tyr Asp Met Ser Glu Lys Leu Val Val Ile Gly Gly
465 470 475 480
Asn Pro Ile Lys Asp Val Thr Lys Glu Glu Asn Leu Tyr Glu Trp Ser
485 490 495
Lys Thr Val Ser Ser Asn Pro Met Pro Ile His Ile Lys Leu Leu Pro
500 505 510
Ile Tyr Lys Ser Phe Asp Ser Glu Glu Leu Lys Glu Ser Tyr Glu Lys
515 520 525
Ala Val Leu Tyr Tyr Thr Arg Leu Tyr Gly Ser Ser Pro His Gly Thr
530 535 540
Ile Gln Lys Asp Glu Asn Asp Ile Ile Lys Ile Leu Thr Ala Ser Thr
545 550 555 560
Thr Ile Thr Lys Ile Gly Ala Pro Pro Ile Thr Ala Glu Cys Pro His
565 570 575
Asn Gln Val Val Leu Phe Gly Tyr Val Leu Lys Gln Asn Phe Trp Asp
580 585 590
Asn Thr Ser Asn Leu Lys Gly Tyr Asp Ile Glu Ile Cys Glu Ala Gly
595 600 605
Leu Asn Ser Cys Thr Ser Lys Gln Gly Ser Thr Asn Lys Tyr Asp Val
610 615 620
Ser Tyr Leu Tyr Ile Glu Cys Gly Thr Gln Ala Met Ser Phe Ser Asp
625 630 635 640
Gln Val Ile Thr Ala Ser Asn Thr Thr Tyr Asn Thr Ile Lys Cys Pro
645 650 655
Asn Asp Tyr Thr Ile Ile Phe Gly Phe Gly Phe Ser Ser Ser Ser Gly
660 665 670
Lys Gly Val Ser Ala Met His Thr His Ile Thr Ser Cys Arg Pro Gly
675 680 685
Met Lys Ser Cys Ser Leu Asn Met Gly Asn Ser Asn Asp Lys Asn Tyr
690 695 700
Met Tyr Leu Val Cys Val Asp Ala Thr Ile Trp Ser Gly Ile Asn Glu
705 710 715 720
Leu Thr Ile Val Ala Lys Asp Asp Phe His Gly Ala Val Asn Arg Ser
725 730 735
Lys Gln Phe Asn Asp Gly Glu Leu Val Leu Ser Cys Gln Glu Asn Gly
740 745 750
Thr Ile Leu Thr Gly Phe Thr Gly Glu Thr His Thr Ser Ser Pro Tyr
755 760 765
Val Lys Ser Pro Phe Ser Lys Cys Leu Lys Ser Leu Lys Ser Cys Ser
770 775 780
Val His Gly Ser Gly Gln Ser Ile Gly Tyr Thr Asn Tyr Lys Ser Leu
785 790 795 800
Phe Ser Ile Ile Leu Cys Lys Asn Gly Glu
805 810
<210> 51
<211> 2538
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> PfSPECT2 人 CO
<400> 51
gccaccatgg ctaagctgag aatcctcaag aagcactatt atgtcgtgtt tatccttctg 60
tatctgtatg acatcagctg cttcaagtgc attcggctga acaaccgttc catctacaag 120
aacaagtaca agaataacgt gcacatagga accaatgaga atatcaggag tattgagaag 180
tatagcaacg tgctgtgtaa cagcatcttg tgtaagaacg ataaaatctc tagcttcatt 240
aaccagagga agaatgtgga tgacgacgat gagtctgaga acgatgacat gtacgagagc 300
accacagctg gctcttctag tgaaacggac aacgagagcg atgaagaaga gaatgacagc 360
agtgacaaca ataatagcga tgaggaacag atagagaact ccaacaacaa taattctgac 420
gaggaacaga atgactcctc ttccaacgac aataatgatg aggagaatga ggaacaggac 480
gacgtcatgg acaatgacca aaacgataag aagatcaagc atagcttcaa tctcgccaac 540
gagagtaaac acactaaaga ggaacgagtg aaagaagaga aaaagctgaa gatctatgac 600
ttcataaacg acaaggagaa aagacttaac tttaatggcg atcagaaaga tgaagataac 660
gaggagaacg atgataaaga tgagaacacg cttgagaatc ggaatatcat ctccaaacac 720
acttcagtgt ttcctggcct gtacttcatc gggattgggt ataacctcct cttcgggaac 780
cccttgggtg aagctgattc ccttatcgat ccaggttatc gggcgcaaat ttacctgatg 840
gaatgggctc tcagcaagga aggcattgcc aacgacctga gcactctgca acccgtgaat 900
ggatggatac gaaaggagaa tgcctgctcc agagttgaat ctattacaga atgcagctct 960
atatctgact acaccaaatc cctgtcagcg gaggcaaagg ttagtggctc ttattgggga 1020
atcgcctcct tctcagcatc caccgggtat agctcttttc tccacgaggt gacaaaacgc 1080
agcaagaaaa ccttcctcgt gaaatccaac tgtgtgaaat acactatcgg gcttcctccc 1140
tatattccct gggacaagac cacggcctac aagaatgccg taaatgaact gccagctgta 1200
tttaccggtt tggataaaga atccgaatgt ccctctgatg tgtacgaaga gaacaagaca 1260
aaatcaaact gcgagaacgt gagtctgtgg atgaagttct tcgacattta tggcactcac 1320
atcatttatg aaagtcaatt gggaggaaag ataacaaaga ttatcaatgt ttccacctca 1380
agtattgagc agatgaagaa aaatggagtc tcagtcaaag cgaaaattca agcacagttt 1440
gggtttggtt cagccggtgg ctcaaccgac gtgaatagca gtaactcctc cgcaaatgat 1500
gagcagtctt acgacatgaa tgaacagctg atcgtcatag gagggaatcc gatcaaagac 1560
gtcaccaagg aggagaatct gtttgagtgg tctaaaactg tgacaaacca tcctatgccg 1620
atcaacatta aactgactcc cattagcgac agttttgact cagacgacct gaaagaatcc 1680
tatgataaag ccatcatcta ctattctcgc ctgtacggac tgtcccctca tgacacaatg 1740
cagaaggatg acaaggatat tatcaagatc ctgaccaacg ctgatacggt taccaagaac 1800
tcagctcctc caatcaacgc tcagtgtcct catgggaaag tcgttatgtt cggattcagc 1860
ctgaagcaga atttctggga caacaccaac gcactcaaag gatacaacat cgaagtctgc 1920
gaagcaggga gcaattcttg cactagcaaa caagggagca gcaataaata cgatacatct 1980
tacctttaca tggaatgtgg cgatcagcca ttgcccttta gcgagcaggt gattagcgag 2040
agtacaagta cctataatac cgtgaaatgt ccgaatgatt actccattct cctgggcttc 2100
ggaatatcaa gtagctcagg gaggataaat agcgctgaat atgtctacag cacaccatgt 2160
attcccggca tgaagtcctg cagcctcaat atgaataatg acaaccagaa gtcatacatc 2220
tacgtgctgt gtgtagatac tacgatctgg agtggcgtga acaacctgag cctggttgcc 2280
cttgatggcg cacatggtaa ggtaaaccgc agtaagaagt actccgacgg tgaactggtt 2340
ggcacctgtc cactggacgg cacagtcctg actggattta aggttgagtt tcacacttca 2400
tctccatatg tgcagacacc tttcgagaaa tgcgccaaaa gcttgaaagc ctgctccgta 2460
catggctccg gtcacgccat tggcattcag aactttaagt cactgttcat atacatgttg 2520
tgcaagaaca ataagtaa 2538
<210> 52
<211> 843
<212> PRT
<213> 恶性疟原虫（Plasmodium falciparum）
<220>
<221> misc_feature
<223> PfSPECT2
<400> 52
Met Ala Lys Leu Arg Ile Leu Lys Lys His Tyr Tyr Val Val Phe Ile
1 5 10 15
Leu Leu Tyr Leu Tyr Asp Ile Ser Cys Phe Lys Cys Ile Arg Leu Asn
20 25 30
Asn Arg Ser Ile Tyr Lys Asn Lys Tyr Lys Asn Asn Val His Ile Gly
35 40 45
Thr Asn Glu Asn Ile Arg Ser Ile Glu Lys Tyr Ser Asn Val Leu Cys
50 55 60
Asn Ser Ile Leu Cys Lys Asn Asp Lys Ile Ser Ser Phe Ile Asn Gln
65 70 75 80
Arg Lys Asn Val Asp Asp Asp Asp Glu Ser Glu Asn Asp Asp Met Tyr
85 90 95
Glu Ser Thr Thr Ala Gly Ser Ser Ser Glu Thr Asp Asn Glu Ser Asp
100 105 110
Glu Glu Glu Asn Asp Ser Ser Asp Asn Asn Asn Ser Asp Glu Glu Gln
115 120 125
Ile Glu Asn Ser Asn Asn Asn Asn Ser Asp Glu Glu Gln Asn Asp Ser
130 135 140
Ser Ser Asn Asp Asn Asn Asp Glu Glu Asn Glu Glu Gln Asp Asp Val
145 150 155 160
Met Asp Asn Asp Gln Asn Asp Lys Lys Ile Lys His Ser Phe Asn Leu
165 170 175
Ala Asn Glu Ser Lys His Thr Lys Glu Glu Arg Val Lys Glu Glu Lys
180 185 190
Lys Leu Lys Ile Tyr Asp Phe Ile Asn Asp Lys Glu Lys Arg Leu Asn
195 200 205
Phe Asn Gly Asp Gln Lys Asp Glu Asp Asn Glu Glu Asn Asp Asp Lys
210 215 220
Asp Glu Asn Thr Leu Glu Asn Arg Asn Ile Ile Ser Lys His Thr Ser
225 230 235 240
Val Phe Pro Gly Leu Tyr Phe Ile Gly Ile Gly Tyr Asn Leu Leu Phe
245 250 255
Gly Asn Pro Leu Gly Glu Ala Asp Ser Leu Ile Asp Pro Gly Tyr Arg
260 265 270
Ala Gln Ile Tyr Leu Met Glu Trp Ala Leu Ser Lys Glu Gly Ile Ala
275 280 285
Asn Asp Leu Ser Thr Leu Gln Pro Val Asn Gly Trp Ile Arg Lys Glu
290 295 300
Asn Ala Cys Ser Arg Val Glu Ser Ile Thr Glu Cys Ser Ser Ile Ser
305 310 315 320
Asp Tyr Thr Lys Ser Leu Ser Ala Glu Ala Lys Val Ser Gly Ser Tyr
325 330 335
Trp Gly Ile Ala Ser Phe Ser Ala Ser Thr Gly Tyr Ser Ser Phe Leu
340 345 350
His Glu Val Thr Lys Arg Ser Lys Lys Thr Phe Leu Val Lys Ser Asn
355 360 365
Cys Val Lys Tyr Thr Ile Gly Leu Pro Pro Tyr Ile Pro Trp Asp Lys
370 375 380
Thr Thr Ala Tyr Lys Asn Ala Val Asn Glu Leu Pro Ala Val Phe Thr
385 390 395 400
Gly Leu Asp Lys Glu Ser Glu Cys Pro Ser Asp Val Tyr Glu Glu Asn
405 410 415
Lys Thr Lys Ser Asn Cys Glu Asn Val Ser Leu Trp Met Lys Phe Phe
420 425 430
Asp Ile Tyr Gly Thr His Ile Ile Tyr Glu Ser Gln Leu Gly Gly Lys
435 440 445
Ile Thr Lys Ile Ile Asn Val Ser Thr Ser Ser Ile Glu Gln Met Lys
450 455 460
Lys Asn Gly Val Ser Val Lys Ala Lys Ile Gln Ala Gln Phe Gly Phe
465 470 475 480
Gly Ser Ala Gly Gly Ser Thr Asp Val Asn Ser Ser Asn Ser Ser Ala
485 490 495
Asn Asp Glu Gln Ser Tyr Asp Met Asn Glu Gln Leu Ile Val Ile Gly
500 505 510
Gly Asn Pro Ile Lys Asp Val Thr Lys Glu Glu Asn Leu Phe Glu Trp
515 520 525
Ser Lys Thr Val Thr Asn His Pro Met Pro Ile Asn Ile Lys Leu Thr
530 535 540
Pro Ile Ser Asp Ser Phe Asp Ser Asp Asp Leu Lys Glu Ser Tyr Asp
545 550 555 560
Lys Ala Ile Ile Tyr Tyr Ser Arg Leu Tyr Gly Leu Ser Pro His Asp
565 570 575
Thr Met Gln Lys Asp Asp Lys Asp Ile Ile Lys Ile Leu Thr Asn Ala
580 585 590
Asp Thr Val Thr Lys Asn Ser Ala Pro Pro Ile Asn Ala Gln Cys Pro
595 600 605
His Gly Lys Val Val Met Phe Gly Phe Ser Leu Lys Gln Asn Phe Trp
610 615 620
Asp Asn Thr Asn Ala Leu Lys Gly Tyr Asn Ile Glu Val Cys Glu Ala
625 630 635 640
Gly Ser Asn Ser Cys Thr Ser Lys Gln Gly Ser Ser Asn Lys Tyr Asp
645 650 655
Thr Ser Tyr Leu Tyr Met Glu Cys Gly Asp Gln Pro Leu Pro Phe Ser
660 665 670
Glu Gln Val Ile Ser Glu Ser Thr Ser Thr Tyr Asn Thr Val Lys Cys
675 680 685
Pro Asn Asp Tyr Ser Ile Leu Leu Gly Phe Gly Ile Ser Ser Ser Ser
690 695 700
Gly Arg Ile Asn Ser Ala Glu Tyr Val Tyr Ser Thr Pro Cys Ile Pro
705 710 715 720
Gly Met Lys Ser Cys Ser Leu Asn Met Asn Asn Asp Asn Gln Lys Ser
725 730 735
Tyr Ile Tyr Val Leu Cys Val Asp Thr Thr Ile Trp Ser Gly Val Asn
740 745 750
Asn Leu Ser Leu Val Ala Leu Asp Gly Ala His Gly Lys Val Asn Arg
755 760 765
Ser Lys Lys Tyr Ser Asp Gly Glu Leu Val Gly Thr Cys Pro Leu Asp
770 775 780
Gly Thr Val Leu Thr Gly Phe Lys Val Glu Phe His Thr Ser Ser Pro
785 790 795 800
Tyr Val Gln Thr Pro Phe Glu Lys Cys Ala Lys Ser Leu Lys Ala Cys
805 810 815
Ser Val His Gly Ser Gly His Ala Ile Gly Ile Gln Asn Phe Lys Ser
820 825 830
Leu Phe Ile Tyr Met Leu Cys Lys Asn Asn Lys
835 840
<210> 53
<211> 842
<212> PRT
<213> 恶性疟原虫（Plasmodium falciparum）
<220>
<221> misc_feature
<223> SPECT2
<400> 53
Met Lys Leu Arg Ile Leu Lys Lys His Tyr Tyr Val Val Phe Ile Leu
1 5 10 15
Leu Tyr Leu Tyr Asp Ile Ser Cys Phe Lys Cys Ile Arg Leu Asn Asn
20 25 30
Arg Ser Ile Tyr Lys Asn Lys Tyr Lys Asn Asn Val His Ile Gly Thr
35 40 45
Asn Glu Asn Ile Arg Ser Ile Glu Lys Tyr Ser Asn Val Leu Cys Asn
50 55 60
Ser Ile Leu Cys Lys Asn Asp Lys Ile Ser Ser Phe Ile Asn Gln Arg
65 70 75 80
Lys Asn Val Asp Asp Asp Asp Glu Ser Glu Asn Asp Asp Met Tyr Glu
85 90 95
Ser Thr Thr Ala Gly Ser Ser Ser Glu Thr Asp Asn Glu Ser Asp Glu
100 105 110
Glu Glu Asn Asp Ser Ser Asp Asn Asn Asn Ser Asp Glu Glu Gln Ile
115 120 125
Glu Asn Ser Asn Asn Asn Asn Ser Asp Glu Glu Gln Asn Asp Ser Ser
130 135 140
Ser Asn Asp Asn Asn Asp Glu Glu Asn Glu Glu Gln Asp Asp Val Met
145 150 155 160
Asp Asn Asp Gln Asn Asp Lys Lys Ile Lys His Ser Phe Asn Leu Ala
165 170 175
Asn Glu Ser Lys His Thr Lys Glu Glu Arg Val Lys Glu Glu Lys Lys
180 185 190
Leu Lys Ile Tyr Asp Phe Ile Asn Asp Lys Glu Lys Arg Leu Asn Phe
195 200 205
Asn Gly Asp Gln Lys Asp Glu Asp Asn Glu Glu Asn Asp Asp Lys Asp
210 215 220
Glu Asn Thr Leu Glu Asn Arg Asn Ile Ile Ser Lys His Thr Ser Val
225 230 235 240
Phe Pro Gly Leu Tyr Phe Ile Gly Ile Gly Tyr Asn Leu Leu Phe Gly
245 250 255
Asn Pro Leu Gly Glu Ala Asp Ser Leu Ile Asp Pro Gly Tyr Arg Ala
260 265 270
Gln Ile Tyr Leu Met Glu Trp Ala Leu Ser Lys Glu Gly Ile Ala Asn
275 280 285
Asp Leu Ser Thr Leu Gln Pro Val Asn Gly Trp Ile Arg Lys Glu Asn
290 295 300
Ala Cys Ser Arg Val Glu Ser Ile Thr Glu Cys Ser Ser Ile Ser Asp
305 310 315 320
Tyr Thr Lys Ser Leu Ser Ala Glu Ala Lys Val Ser Gly Ser Tyr Trp
325 330 335
Gly Ile Ala Ser Phe Ser Ala Ser Thr Gly Tyr Ser Ser Phe Leu His
340 345 350
Glu Val Thr Lys Arg Ser Lys Lys Thr Phe Leu Val Lys Ser Asn Cys
355 360 365
Val Lys Tyr Thr Ile Gly Leu Pro Pro Tyr Ile Pro Trp Asp Lys Thr
370 375 380
Thr Ala Tyr Lys Asn Ala Val Asn Glu Leu Pro Ala Val Phe Thr Gly
385 390 395 400
Leu Asp Lys Glu Ser Glu Cys Pro Ser Asp Val Tyr Glu Glu Asn Lys
405 410 415
Thr Lys Ser Asn Cys Glu Asn Val Ser Leu Trp Met Lys Phe Phe Asp
420 425 430
Ile Tyr Gly Thr His Ile Ile Tyr Glu Ser Gln Leu Gly Gly Lys Ile
435 440 445
Thr Lys Ile Ile Asn Val Ser Thr Ser Ser Ile Glu Gln Met Lys Lys
450 455 460
Asn Gly Val Ser Val Lys Ala Lys Ile Gln Ala Gln Phe Gly Phe Gly
465 470 475 480
Ser Ala Gly Gly Ser Thr Asp Val Asn Ser Ser Asn Ser Ser Ala Asn
485 490 495
Asp Glu Gln Ser Tyr Asp Met Asn Glu Gln Leu Ile Val Ile Gly Gly
500 505 510
Asn Pro Ile Lys Asp Val Thr Lys Glu Glu Asn Leu Phe Glu Trp Ser
515 520 525
Lys Thr Val Thr Asn His Pro Met Pro Ile Asn Ile Lys Leu Thr Pro
530 535 540
Ile Ser Asp Ser Phe Asp Ser Asp Asp Leu Lys Glu Ser Tyr Asp Lys
545 550 555 560
Ala Ile Ile Tyr Tyr Ser Arg Leu Tyr Gly Leu Ser Pro His Asp Thr
565 570 575
Met Gln Lys Asp Asp Lys Asp Ile Ile Lys Ile Leu Thr Asn Ala Asp
580 585 590
Thr Val Thr Lys Asn Ser Ala Pro Pro Ile Asn Ala Gln Cys Pro His
595 600 605
Gly Lys Val Val Met Phe Gly Phe Ser Leu Lys Gln Asn Phe Trp Asp
610 615 620
Asn Thr Asn Ala Leu Lys Gly Tyr Asn Ile Glu Val Cys Glu Ala Gly
625 630 635 640
Ser Asn Ser Cys Thr Ser Lys Gln Gly Ser Ser Asn Lys Tyr Asp Thr
645 650 655
Ser Tyr Leu Tyr Met Glu Cys Gly Asp Gln Pro Leu Pro Phe Ser Glu
660 665 670
Gln Val Ile Ser Glu Ser Thr Ser Thr Tyr Asn Thr Val Lys Cys Pro
675 680 685
Asn Asp Tyr Ser Ile Leu Leu Gly Phe Gly Ile Ser Ser Ser Ser Gly
690 695 700
Arg Ile Asn Ser Ala Glu Tyr Val Tyr Ser Thr Pro Cys Ile Pro Gly
705 710 715 720
Met Lys Ser Cys Ser Leu Asn Met Asn Asn Asp Asn Gln Lys Ser Tyr
725 730 735
Ile Tyr Val Leu Cys Val Asp Thr Thr Ile Trp Ser Gly Val Asn Asn
740 745 750
Leu Ser Leu Val Ala Leu Asp Gly Ala His Gly Lys Val Asn Arg Ser
755 760 765
Lys Lys Tyr Ser Asp Gly Glu Leu Val Gly Thr Cys Pro Leu Asp Gly
770 775 780
Thr Val Leu Thr Gly Phe Lys Val Glu Phe His Thr Ser Ser Pro Tyr
785 790 795 800
Val Gln Thr Pro Phe Glu Lys Cys Ala Lys Ser Leu Lys Ala Cys Ser
805 810 815
Val His Gly Ser Gly His Ala Ile Gly Ile Gln Asn Phe Lys Ser Leu
820 825 830
Phe Ile Tyr Met Leu Cys Lys Asn Asn Lys
835 840
<210> 54
<211> 2541
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 合成序列
<220>
<221> misc_feature
<223> PvSPECT2 人 CO
<400> 54
gccaccatgg ctaagccaag aaatataaac tctctgttgg ccatatggtg tattctcttc 60
tcaatctgtg agtacggcta tgtgggatct ttgagaatag gtcttcggag aaactatagt 120
caaggggact cctctgatcg gctcgggggt agtgacgtgg acgtgagacg gtccggcgag 180
cttatggacc tcggtaagaa cgcaaacatc ctctgcaata acatcagctg taacagtaaa 240
aaagaggctt cttttctgag tcagaaaaaa caacttgagg atgacgatga cgacgagctg 300
gccgcgctgt acaatgatga tgatgacgat gctagcacaa ccactggggg gggttcagaa 360
actagcgatg acgacgagct tgaactccct gatcaggacg aaggtgcgga cgagggtgaa 420
gcagaggacc aactcgcgac acaggaggat tccgacaact cagggaccga ccaaggtctg 480
aaaaaaaaag taaaccttag ccgccacgaa aagttgatag aagacaaaaa acaacagaca 540
gagaatactt ttaagaagta caggtttggg gatgaggagg aagagtcaga ggagaaatcc 600
ccggggaaat ctaagtcact cgatccaagc agcctcgatg atgacgacgg cgaaggtgat 660
gatgatgatg atggcgacga gagagaaaag aagcaaagta atacaaggaa agccatgaag 720
aaggacttgg atgttttccc agggctttac tttgcaggca tcggatacga tagcctcttc 780
ggaaaccccc tgggcgaggc tgacagcttg accgatcccg ggtaccgggg ccaaatcata 840
ctcatgaact gggaactctc aaataaggga gttgcgaatg atcttgcgac attgcaacct 900
ctgaatggat ggatccgcaa ggaaaatgcg tgcagtcgag cagaatctat aaaagagtgt 960
tcttcagtat cagactacac gaaaaatttg accgcagagg catctgtatc tggctcctac 1020
atgggctttg gagcttttag tgctagtact ggttataaga aatttttgca agaagccagt 1080
aaacgcacgt ccaaaactta tctggtaaaa agcaattgcg tgaaatacac ggtcgggctc 1140
cctccttatg tgcgctggga gcagacgacc gcctttaaga acgctgtgaa cgggttgcca 1200
ccgcatttca ccggactgga ggccgactct gaatgcgcct ctgatgtcta cgaacaaaag 1260
aagacatctg aagaatgcga aaccgtacat gcctggatac gcttttttaa gacttatgga 1320
acacacgtga tcatggaagc acagcttggc ggcaaaataa caaagattat ccgggtcgag 1380
aatagctccg tcaatcaaat gaagaaagac ggagtcagcg tgaaagccca gatcaaagcc 1440
caattcggct ttgcatctgt ggggggaagc acaaacgtct ctagtgacca tagttcaaaa 1500
aagaacgagg ataattatga gatgtctgag cagctggtag tgattggagg caacccaatc 1560
aaggatgtga cgaaagaaga aaatctctat gagtggtcta aaacggtctc aacaaatccg 1620
atgcctatca atatcaagct cctgccgatt agcactatct ttgactccga cgatctgaaa 1680
aatagttacg aaaaggcatt gatctactac actcgcttgt atggcttttc accccatgat 1740
acaatgcaaa aagatgaaaa ggatattgtt aagatcctga cggccagcac cacagtcaca 1800
aagacaggcc cgccgcctat atctgcagag tgtccgcata atatggtggt cttgttcggt 1860
ttcgtagtca aacagaattt ttgggatcat acgaacaaac tccagagtta tgaaatggag 1920
atttgtgaaa gcggtgccag ctcatgcacg tctaagcagg gaaatacaaa taagtatgac 1980
gtatcctata cgtatattga gtgcggacca caggcattgc cgttcactga gcaggttgta 2040
tctgtatccg gtacaacata taactctgtt aaatgcccga atgactacag cgtgttgttt 2100
ggctttggaa tggctacgag ttctggcagg caccaaagtg cgctttatag ctatttcaca 2160
ccatgtcgac cagggctcaa aagctgtagc ttgaatatga acgaacatga tgataagtct 2220
tacatttacc tggtctgcgt cgacgcgact atctggacgg gacttaacgc gttgagcatg 2280
atcgcgaaag acgatttgca cagtgccgtg aaccggtacc aacaattcaa tgatggagag 2340
ctggttgtga cgtgtcctag cgagggcact atactgactg gattctatgg ggagacccat 2400
acctccagtc catatgttac tgtacctttt gggaaatgcg caaagtcatt gaaagcctgt 2460
tctgttcatg ggtccggcca agccataggc attcacaatt atagaacttt gttcacagtc 2520
gcactgtgta agaataatta g 2541
<210> 55
<211> 844
<212> PRT
<213> 间日疟原虫（Plasmodium vivax）
<220>
<221> misc_feature
<223> PvSPECT2
<400> 55
Met Ala Lys Pro Arg Asn Ile Asn Ser Leu Leu Ala Ile Trp Cys Ile
1 5 10 15
Leu Phe Ser Ile Cys Glu Tyr Gly Tyr Val Gly Ser Leu Arg Ile Gly
20 25 30
Leu Arg Arg Asn Tyr Ser Gln Gly Asp Ser Ser Asp Arg Leu Gly Gly
35 40 45
Ser Asp Val Asp Val Arg Arg Ser Gly Glu Leu Met Asp Leu Gly Lys
50 55 60
Asn Ala Asn Ile Leu Cys Asn Asn Ile Ser Cys Asn Ser Lys Lys Glu
65 70 75 80
Ala Ser Phe Leu Ser Gln Lys Lys Gln Leu Glu Asp Asp Asp Asp Asp
85 90 95
Glu Leu Ala Ala Leu Tyr Asn Asp Asp Asp Asp Asp Ala Ser Thr Thr
100 105 110
Thr Gly Gly Gly Ser Glu Thr Ser Asp Asp Asp Glu Leu Glu Leu Pro
115 120 125
Asp Gln Asp Glu Gly Ala Asp Glu Gly Glu Ala Glu Asp Gln Leu Ala
130 135 140
Thr Gln Glu Asp Ser Asp Asn Ser Gly Thr Asp Gln Gly Leu Lys Lys
145 150 155 160
Lys Val Asn Leu Ser Arg His Glu Lys Leu Ile Glu Asp Lys Lys Gln
165 170 175
Gln Thr Glu Asn Thr Phe Lys Lys Tyr Arg Phe Gly Asp Glu Glu Glu
180 185 190
Glu Ser Glu Glu Lys Ser Pro Gly Lys Ser Lys Ser Leu Asp Pro Ser
195 200 205
Ser Leu Asp Asp Asp Asp Gly Glu Gly Asp Asp Asp Asp Asp Gly Asp
210 215 220
Glu Arg Glu Lys Lys Gln Ser Asn Thr Arg Lys Ala Met Lys Lys Asp
225 230 235 240
Leu Asp Val Phe Pro Gly Leu Tyr Phe Ala Gly Ile Gly Tyr Asp Ser
245 250 255
Leu Phe Gly Asn Pro Leu Gly Glu Ala Asp Ser Leu Thr Asp Pro Gly
260 265 270
Tyr Arg Gly Gln Ile Ile Leu Met Asn Trp Glu Leu Ser Asn Lys Gly
275 280 285
Val Ala Asn Asp Leu Ala Thr Leu Gln Pro Leu Asn Gly Trp Ile Arg
290 295 300
Lys Glu Asn Ala Cys Ser Arg Ala Glu Ser Ile Lys Glu Cys Ser Ser
305 310 315 320
Val Ser Asp Tyr Thr Lys Asn Leu Thr Ala Glu Ala Ser Val Ser Gly
325 330 335
Ser Tyr Met Gly Phe Gly Ala Phe Ser Ala Ser Thr Gly Tyr Lys Lys
340 345 350
Phe Leu Gln Glu Ala Ser Lys Arg Thr Ser Lys Thr Tyr Leu Val Lys
355 360 365
Ser Asn Cys Val Lys Tyr Thr Val Gly Leu Pro Pro Tyr Val Arg Trp
370 375 380
Glu Gln Thr Thr Ala Phe Lys Asn Ala Val Asn Gly Leu Pro Pro His
385 390 395 400
Phe Thr Gly Leu Glu Ala Asp Ser Glu Cys Ala Ser Asp Val Tyr Glu
405 410 415
Gln Lys Lys Thr Ser Glu Glu Cys Glu Thr Val His Ala Trp Ile Arg
420 425 430
Phe Phe Lys Thr Tyr Gly Thr His Val Ile Met Glu Ala Gln Leu Gly
435 440 445
Gly Lys Ile Thr Lys Ile Ile Arg Val Glu Asn Ser Ser Val Asn Gln
450 455 460
Met Lys Lys Asp Gly Val Ser Val Lys Ala Gln Ile Lys Ala Gln Phe
465 470 475 480
Gly Phe Ala Ser Val Gly Gly Ser Thr Asn Val Ser Ser Asp His Ser
485 490 495
Ser Lys Lys Asn Glu Asp Asn Tyr Glu Met Ser Glu Gln Leu Val Val
500 505 510
Ile Gly Gly Asn Pro Ile Lys Asp Val Thr Lys Glu Glu Asn Leu Tyr
515 520 525
Glu Trp Ser Lys Thr Val Ser Thr Asn Pro Met Pro Ile Asn Ile Lys
530 535 540
Leu Leu Pro Ile Ser Thr Ile Phe Asp Ser Asp Asp Leu Lys Asn Ser
545 550 555 560
Tyr Glu Lys Ala Leu Ile Tyr Tyr Thr Arg Leu Tyr Gly Phe Ser Pro
565 570 575
His Asp Thr Met Gln Lys Asp Glu Lys Asp Ile Val Lys Ile Leu Thr
580 585 590
Ala Ser Thr Thr Val Thr Lys Thr Gly Pro Pro Pro Ile Ser Ala Glu
595 600 605
Cys Pro His Asn Met Val Val Leu Phe Gly Phe Val Val Lys Gln Asn
610 615 620
Phe Trp Asp His Thr Asn Lys Leu Gln Ser Tyr Glu Met Glu Ile Cys
625 630 635 640
Glu Ser Gly Ala Ser Ser Cys Thr Ser Lys Gln Gly Asn Thr Asn Lys
645 650 655
Tyr Asp Val Ser Tyr Thr Tyr Ile Glu Cys Gly Pro Gln Ala Leu Pro
660 665 670
Phe Thr Glu Gln Val Val Ser Val Ser Gly Thr Thr Tyr Asn Ser Val
675 680 685
Lys Cys Pro Asn Asp Tyr Ser Val Leu Phe Gly Phe Gly Met Ala Thr
690 695 700
Ser Ser Gly Arg His Gln Ser Ala Leu Tyr Ser Tyr Phe Thr Pro Cys
705 710 715 720
Arg Pro Gly Leu Lys Ser Cys Ser Leu Asn Met Asn Glu His Asp Asp
725 730 735
Lys Ser Tyr Ile Tyr Leu Val Cys Val Asp Ala Thr Ile Trp Thr Gly
740 745 750
Leu Asn Ala Leu Ser Met Ile Ala Lys Asp Asp Leu His Ser Ala Val
755 760 765
Asn Arg Tyr Gln Gln Phe Asn Asp Gly Glu Leu Val Val Thr Cys Pro
770 775 780
Ser Glu Gly Thr Ile Leu Thr Gly Phe Tyr Gly Glu Thr His Thr Ser
785 790 795 800
Ser Pro Tyr Val Thr Val Pro Phe Gly Lys Cys Ala Lys Ser Leu Lys
805 810 815
Ala Cys Ser Val His Gly Ser Gly Gln Ala Ile Gly Ile His Asn Tyr
820 825 830
Arg Thr Leu Phe Thr Val Ala Leu Cys Lys Asn Asn
835 840
<210> 56
<211> 843
<212> PRT
<213> 间日疟原虫（Plasmodium vivax）
<220>
<221> misc_feature
<223> SPECT2
<400> 56
Met Lys Pro Arg Asn Ile Asn Ser Leu Leu Ala Ile Trp Cys Ile Leu
1 5 10 15
Phe Ser Ile Cys Glu Tyr Gly Tyr Val Gly Ser Leu Arg Ile Gly Leu
20 25 30
Arg Arg Asn Tyr Ser Gln Gly Asp Ser Ser Asp Arg Leu Gly Gly Ser
35 40 45
Asp Val Asp Val Arg Arg Ser Gly Glu Leu Met Asp Phe Gly Lys Asn
50 55 60
Ala Asn Ile Leu Cys Asn Asn Ile Ser Cys Asn Ser Lys Lys Glu Ala
65 70 75 80
Ser Phe Leu Ser Gln Lys Lys Gln Leu Glu Asp Asp Asp Asp Asp Glu
85 90 95
Leu Ala Ala Leu Tyr Asn Asp Asp Asp Asp Asp Ala Ser Thr Thr Thr
100 105 110
Gly Gly Gly Ser Glu Thr Ser Asp Asp Asp Glu Leu Glu Leu Pro Asp
115 120 125
Gln Asp Glu Gly Ala Asp Glu Gly Glu Ala Glu Asp Gln Leu Ala Thr
130 135 140
Gln Glu Asp Ser Asp Asn Ser Gly Thr Asp Gln Gly Leu Lys Lys Lys
145 150 155 160
Val Ser Leu Ser Arg His Glu Lys Leu Ile Glu Asp Lys Lys Gln Gln
165 170 175
Thr Glu Asn Thr Phe Lys Lys Tyr Arg Phe Gly Asp Glu Glu Glu Glu
180 185 190
Ser Glu Glu Lys Ser Pro Gly Lys Ser Lys Ser Leu Asp Pro Ser Ser
195 200 205
Leu Asp Asp Asp Asp Gly Glu Gly Asp Asp Asp Asp Asp Gly Asp Glu
210 215 220
Arg Glu Lys Lys Gln Ser Asn Thr Arg Lys Ala Met Lys Lys Asp Leu
225 230 235 240
Asp Val Phe Pro Gly Leu Tyr Phe Ala Gly Ile Gly Tyr Asp Ser Leu
245 250 255
Phe Gly Asn Pro Leu Gly Glu Ala Asp Ser Leu Thr Asp Pro Gly Tyr
260 265 270
Arg Gly Gln Ile Ile Leu Met Asn Trp Glu Leu Ser Asn Lys Gly Val
275 280 285
Ala Asn Asp Leu Ala Thr Leu Gln Pro Leu Asn Gly Trp Ile Arg Lys
290 295 300
Glu Asn Ala Cys Ser Arg Ala Glu Ser Ile Lys Glu Cys Ser Ser Val
305 310 315 320
Ser Asp Tyr Thr Lys Asn Leu Thr Ala Glu Ala Ser Val Ser Gly Ser
325 330 335
Tyr Met Gly Phe Gly Ala Phe Ser Ala Ser Thr Gly Tyr Lys Lys Phe
340 345 350
Leu Gln Glu Ala Ser Lys Arg Thr Ser Lys Thr Tyr Leu Val Lys Ser
355 360 365
Asn Cys Val Lys Tyr Thr Val Gly Leu Pro Pro Tyr Val Arg Trp Glu
370 375 380
Gln Thr Thr Ala Phe Lys Asn Ala Val Asn Gly Leu Pro Pro His Phe
385 390 395 400
Thr Gly Leu Glu Ala Asp Ser Glu Cys Ala Ser Asp Val Tyr Glu Gln
405 410 415
Lys Lys Thr Ser Glu Glu Cys Glu Thr Val His Ala Trp Ile Arg Phe
420 425 430
Phe Lys Thr Tyr Gly Thr His Val Ile Met Glu Ala Gln Leu Gly Gly
435 440 445
Lys Ile Thr Lys Ile Ile Arg Val Glu Asn Ser Ser Val Asn Gln Met
450 455 460
Lys Lys Asp Gly Val Ser Val Lys Ala Gln Ile Lys Ala Gln Phe Gly
465 470 475 480
Phe Ala Ser Val Gly Gly Ser Thr Asn Val Ser Ser Asp His Ser Ser
485 490 495
Lys Lys Asn Glu Asp Asn Tyr Glu Met Ser Glu Gln Leu Val Val Ile
500 505 510
Gly Gly Asn Pro Ile Lys Asp Val Thr Lys Glu Glu Asn Leu Tyr Glu
515 520 525
Trp Ser Lys Thr Val Ser Thr Asn Pro Met Pro Ile Asn Ile Lys Leu
530 535 540
Leu Pro Ile Ser Thr Ile Phe Asp Ser Asp Asp Leu Lys Asn Ser Tyr
545 550 555 560
Glu Lys Ala Leu Ile Tyr Tyr Thr Arg Leu Tyr Gly Phe Ser Pro His
565 570 575
Asp Thr Met Gln Lys Asp Glu Lys Asp Ile Val Lys Ile Leu Thr Ala
580 585 590
Ser Thr Thr Val Thr Lys Thr Gly Pro Pro Pro Ile Ser Ala Glu Cys
595 600 605
Pro His Asn Met Val Val Leu Phe Gly Phe Val Val Lys Gln Asn Phe
610 615 620
Trp Asp His Thr Asn Lys Leu Gln Ser Tyr Glu Met Glu Ile Cys Glu
625 630 635 640
Ser Gly Ala Ser Ser Cys Thr Ser Lys Gln Gly Asn Thr Asn Lys Tyr
645 650 655
Asp Val Ser Tyr Thr Tyr Ile Glu Cys Gly Pro Gln Ala Leu Pro Phe
660 665 670
Thr Glu Gln Val Val Ser Val Ser Gly Thr Thr Tyr Asn Ser Val Lys
675 680 685
Cys Pro Asn Asp Tyr Ser Val Leu Phe Gly Phe Gly Met Ala Thr Ser
690 695 700
Ser Gly Arg His Gln Ser Ala Leu Tyr Ser Tyr Phe Thr Pro Cys Arg
705 710 715 720
Pro Gly Leu Lys Ser Cys Ser Leu Asn Met Asn Glu His Asp Asp Lys
725 730 735
Ser Tyr Ile Tyr Leu Val Cys Val Asp Ala Thr Ile Trp Thr Gly Leu
740 745 750
Asn Ala Leu Ser Met Ile Ala Lys Asp Asp Leu His Ser Ala Val Asn
755 760 765
Arg Tyr Gln Gln Phe Asn Asp Gly Glu Leu Val Val Thr Cys Pro Ser
770 775 780
Glu Gly Thr Ile Leu Thr Gly Phe Tyr Gly Glu Thr His Thr Ser Ser
785 790 795 800
Pro Tyr Val Thr Val Pro Phe Gly Lys Cys Ala Lys Ser Leu Lys Ala
805 810 815
Cys Ser Val His Gly Ser Gly Gln Ala Ile Gly Ile His Asn Tyr Arg
820 825 830
Thr Leu Phe Thr Val Ala Leu Cys Lys Asn Asn
835 840
<210> 57
<211> 2478
<212> DNA
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> GPI_P113
<400> 57
ggatccgcca ccatggctaa gatcttcctg tttagtttct tcttcgtatg gttccagtat 60
tgctattcta agaacccctc agactacgct catagcattg tctctaattt tgagtccgag 120
aatacgttga agtgtttgaa aggcaacgtg tatatacttc aatgtcagat caagtgcatg 180
aactcaaaca acgagatcat atacaaggag tgtctcaacg atatcgaaaa gatttgcaaa 240
gataagaaaa catgcagtta ttacttcgac tacatcttca aaaccaagaa tcacaagctt 300
agaaacaaca acaataacaa catctacatc gacaattgca tcgattccga caagaatgag 360
atcaaatcta ccttcacctg tgtcctcaat ccattgctgg aatttgacaa taacaaccac 420
gtcatttata acttcctgct gaacaacaag aacaatgaca agatcgtctg taaaaatagc 480
aacatatata ttaacaacgc tactattcac tataccttct ccgacattaa gtttaaagac 540
gtgactagct acataaaaga gaaatgtaac gagaaaacta attgcgtgat aaacccctat 600
tctatccaaa ccgatatcct gaatgagaag aacgacgcat atctgctgaa ctcatacatc 660
agcatatcct tcgcatgtgt gaaaattaac ctggaaagct atctgtacgg aggtgatatc 720
gatgaatttg atcagataaa cgatgaagaa aatgaggaca ataagtacct ggatcacaat 780
gatctcgatg aaaaaaatga ggaaattatt tccctgaaga acgaaatcaa cgacatcctg 840
aatgacgaga agatcgataa catcgccgag aagctgaaga ttgcgaaatt tacaataagc 900
aagaaaatca acgaggagat caagaagaag aacgacatct ttaacaacct ggccaacgac 960
atctaccagt tcatcggtaa cgagtactac tttacttccg acatcaaaga catgattgaa 1020
gataggtaca atgaactgaa caaaacatct cagtctgact tgtattacat ttaccttctg 1080
aatgtgtttg acattgaaaa gatttacggg atctacctgt ctagttatca ggagcgactg 1140
cagcagatcc tgcaaaccaa catgacaaat ctggactatg ttgagaagaa gattggaagc 1200
ttgcggaaca tttacatgtt cctgtataaa aagagtaaga agtataacgc gctggatatc 1260
ttcgacgaat actacgatta tgtgctcaat tacaatgact tcgctaaaga caatgagata 1320
atcagcgccg acattttcat taagtcaaaa cctgatatcc ctcaactcaa tttcgagatc 1380
aacaatgaga acaagaacgt gaagtataaa gacgttaccg atcttgacga gctggataac 1440
ctcaacagga taaatagaat tatcaatatc cggaatgtgc tggtcaagca gctcaagatc 1500
ctttacaacc agagaaataa catatttatc aaacaggcca tgctggtgaa atcatactgc 1560
tacaaaaatc cactggatat tacggatttc agcagcatct tcaagaataa ctacaataag 1620
ttgaaatacg acgcctataa ggagggcaat ggacacatta acgtggccga caaaatcaat 1680
ccaaactttg tcgtgaaata cctgaacaat ctttataaac agcatgttaa caagaattat 1740
atcctgaaca gctgggaccc taaatacaat cgcatgaata agaagattaa gatcattctc 1800
attttggggt atggccaggt aatccagatc gagaaacaga ttaaccgtca tataggcaag 1860
tacaacgccc tgcttgaaaa ggcaaccctc tataacgtgg gaaatctctt tacacagact 1920
acgaatattt tgaacgacat ctcaggctca ctgaacgatg ggcttgaccc gaacatccat 1980
gatcaggagg atgtcactgt cgttgaatcc tctgaaagta acaagctcgc agaacctgag 2040
gaacccattg agaaggtaga ggttgatagg gtagagaaat ccgatgatgc caataatgcc 2100
actcaacaag tgacaggaac agacgaggcc aattacgata cagctagcgg gaatagtacc 2160
aacatcaaca ttgacaacat agactatggt gtggacgaat ctatccgaat tattaagtac 2220
tccaaggctg aggaggatga atataatgag agcggcaata acgaaaatga aaacaacgag 2280
aataacgaga acgagaacaa tgaaaatgag aacaacgaga atgagaataa tgagaatgaa 2340
aataacgaga acgaaaatat agaactgaag aatatagaac acgaaaacaa atccaacgca 2400
tctagcgctt ccctcagtaa cattttcttt acctttatca ttgcagctct gtttatccgc 2460
ccctttctgt gactcgag 2478
<210> 58
<211> 819
<212> PRT
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> GPI_P113
<400> 58
Met Ala Lys Ile Phe Leu Phe Ser Phe Phe Phe Val Trp Phe Gln Tyr
1 5 10 15
Cys Tyr Ser Lys Asn Pro Ser Asp Tyr Ala His Ser Ile Val Ser Asn
20 25 30
Phe Glu Ser Glu Asn Thr Leu Lys Cys Leu Lys Gly Asn Val Tyr Ile
35 40 45
Leu Gln Cys Gln Ile Lys Cys Met Asn Ser Asn Asn Glu Ile Ile Tyr
50 55 60
Lys Glu Cys Leu Asn Asp Ile Glu Lys Ile Cys Lys Asp Lys Lys Thr
65 70 75 80
Cys Ser Tyr Tyr Phe Asp Tyr Ile Phe Lys Thr Lys Asn His Lys Leu
85 90 95
Arg Asn Asn Asn Asn Asn Asn Ile Tyr Ile Asp Asn Cys Ile Asp Ser
100 105 110
Asp Lys Asn Glu Ile Lys Ser Thr Phe Thr Cys Val Leu Asn Pro Leu
115 120 125
Leu Glu Phe Asp Asn Asn Asn His Val Ile Tyr Asn Phe Leu Leu Asn
130 135 140
Asn Lys Asn Asn Asp Lys Ile Val Cys Lys Asn Ser Asn Ile Tyr Ile
145 150 155 160
Asn Asn Ala Thr Ile His Tyr Thr Phe Ser Asp Ile Lys Phe Lys Asp
165 170 175
Val Thr Ser Tyr Ile Lys Glu Lys Cys Asn Glu Lys Thr Asn Cys Val
180 185 190
Ile Asn Pro Tyr Ser Ile Gln Thr Asp Ile Leu Asn Glu Lys Asn Asp
195 200 205
Ala Tyr Leu Leu Asn Ser Tyr Ile Ser Ile Ser Phe Ala Cys Val Lys
210 215 220
Ile Asn Leu Glu Ser Tyr Leu Tyr Gly Gly Asp Ile Asp Glu Phe Asp
225 230 235 240
Gln Ile Asn Asp Glu Glu Asn Glu Asp Asn Lys Tyr Leu Asp His Asn
245 250 255
Asp Leu Asp Glu Lys Asn Glu Glu Ile Ile Ser Leu Lys Asn Glu Ile
260 265 270
Asn Asp Ile Leu Asn Asp Glu Lys Ile Asp Asn Ile Ala Glu Lys Leu
275 280 285
Lys Ile Ala Lys Phe Thr Ile Ser Lys Lys Ile Asn Glu Glu Ile Lys
290 295 300
Lys Lys Asn Asp Ile Phe Asn Asn Leu Ala Asn Asp Ile Tyr Gln Phe
305 310 315 320
Ile Gly Asn Glu Tyr Tyr Phe Thr Ser Asp Ile Lys Asp Met Ile Glu
325 330 335
Asp Arg Tyr Asn Glu Leu Asn Lys Thr Ser Gln Ser Asp Leu Tyr Tyr
340 345 350
Ile Tyr Leu Leu Asn Val Phe Asp Ile Glu Lys Ile Tyr Gly Ile Tyr
355 360 365
Leu Ser Ser Tyr Gln Glu Arg Leu Gln Gln Ile Leu Gln Thr Asn Met
370 375 380
Thr Asn Leu Asp Tyr Val Glu Lys Lys Ile Gly Ser Leu Arg Asn Ile
385 390 395 400
Tyr Met Phe Leu Tyr Lys Lys Ser Lys Lys Tyr Asn Ala Leu Asp Ile
405 410 415
Phe Asp Glu Tyr Tyr Asp Tyr Val Leu Asn Tyr Asn Asp Phe Ala Lys
420 425 430
Asp Asn Glu Ile Ile Ser Ala Asp Ile Phe Ile Lys Ser Lys Pro Asp
435 440 445
Ile Pro Gln Leu Asn Phe Glu Ile Asn Asn Glu Asn Lys Asn Val Lys
450 455 460
Tyr Lys Asp Val Thr Asp Leu Asp Glu Leu Asp Asn Leu Asn Arg Ile
465 470 475 480
Asn Arg Ile Ile Asn Ile Arg Asn Val Leu Val Lys Gln Leu Lys Ile
485 490 495
Leu Tyr Asn Gln Arg Asn Asn Ile Phe Ile Lys Gln Ala Met Leu Val
500 505 510
Lys Ser Tyr Cys Tyr Lys Asn Pro Leu Asp Ile Thr Asp Phe Ser Ser
515 520 525
Ile Phe Lys Asn Asn Tyr Asn Lys Leu Lys Tyr Asp Ala Tyr Lys Glu
530 535 540
Gly Asn Gly His Ile Asn Val Ala Asp Lys Ile Asn Pro Asn Phe Val
545 550 555 560
Val Lys Tyr Leu Asn Asn Leu Tyr Lys Gln His Val Asn Lys Asn Tyr
565 570 575
Ile Leu Asn Ser Trp Asp Pro Lys Tyr Asn Arg Met Asn Lys Lys Ile
580 585 590
Lys Ile Ile Leu Ile Leu Gly Tyr Gly Gln Val Ile Gln Ile Glu Lys
595 600 605
Gln Ile Asn Arg His Ile Gly Lys Tyr Asn Ala Leu Leu Glu Lys Ala
610 615 620
Thr Leu Tyr Asn Val Gly Asn Leu Phe Thr Gln Thr Thr Asn Ile Leu
625 630 635 640
Asn Asp Ile Ser Gly Ser Leu Asn Asp Gly Leu Asp Pro Asn Ile His
645 650 655
Asp Gln Glu Asp Val Thr Val Val Glu Ser Ser Glu Ser Asn Lys Leu
660 665 670
Ala Glu Pro Glu Glu Pro Ile Glu Lys Val Glu Val Asp Arg Val Glu
675 680 685
Lys Ser Asp Asp Ala Asn Asn Ala Thr Gln Gln Val Thr Gly Thr Asp
690 695 700
Glu Ala Asn Tyr Asp Thr Ala Ser Gly Asn Ser Thr Asn Ile Asn Ile
705 710 715 720
Asp Asn Ile Asp Tyr Gly Val Asp Glu Ser Ile Arg Ile Ile Lys Tyr
725 730 735
Ser Lys Ala Glu Glu Asp Glu Tyr Asn Glu Ser Gly Asn Asn Glu Asn
740 745 750
Glu Asn Asn Glu Asn Asn Glu Asn Glu Asn Asn Glu Asn Glu Asn Asn
755 760 765
Glu Asn Glu Asn Asn Glu Asn Glu Asn Asn Glu Asn Glu Asn Ile Glu
770 775 780
Leu Lys Asn Ile Glu His Glu Asn Lys Ser Asn Ala Ser Ser Ala Ser
785 790 795 800
Leu Ser Asn Ile Phe Phe Thr Phe Ile Ile Ala Ala Leu Phe Ile Arg
805 810 815
Pro Phe Leu
<210> 59
<211> 818
<212> PRT
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> P113
<400> 59
Met Lys Ile Phe Leu Phe Ser Phe Phe Phe Val Trp Phe Gln Tyr Cys
1 5 10 15
Tyr Ser Lys Asn Pro Ser Asp Tyr Ala His Ser Ile Val Ser Asn Phe
20 25 30
Glu Ser Glu Asn Thr Leu Lys Cys Leu Lys Gly Asn Val Tyr Ile Leu
35 40 45
Gln Cys Gln Ile Lys Cys Met Asn Ser Asn Asn Glu Ile Ile Tyr Lys
50 55 60
Glu Cys Leu Asn Asp Ile Glu Lys Ile Cys Lys Asp Lys Lys Thr Cys
65 70 75 80
Ser Tyr Tyr Phe Asp Tyr Ile Phe Lys Thr Lys Asn His Lys Leu Arg
85 90 95
Asn Asn Asn Asn Asn Asn Ile Tyr Ile Asp Asn Cys Ile Asp Ser Asp
100 105 110
Lys Asn Glu Ile Lys Ser Thr Phe Thr Cys Val Leu Asn Pro Leu Leu
115 120 125
Glu Phe Asp Asn Asn Asn His Val Ile Tyr Asn Phe Leu Leu Asn Asn
130 135 140
Lys Asn Asn Asp Lys Ile Val Cys Lys Asn Ser Asn Ile Tyr Ile Asn
145 150 155 160
Asn Ala Thr Ile His Tyr Thr Phe Ser Asp Ile Lys Phe Lys Asp Val
165 170 175
Thr Ser Tyr Ile Lys Glu Lys Cys Asn Glu Lys Thr Asn Cys Val Ile
180 185 190
Asn Pro Tyr Ser Ile Gln Thr Asp Ile Leu Asn Glu Lys Asn Asp Ala
195 200 205
Tyr Leu Leu Asn Ser Tyr Ile Ser Ile Ser Phe Ala Cys Val Lys Ile
210 215 220
Asn Leu Glu Ser Tyr Leu Tyr Gly Gly Asp Ile Asp Glu Phe Asp Gln
225 230 235 240
Ile Asn Asp Glu Glu Asn Glu Asp Asn Lys Tyr Leu Asp His Asn Asp
245 250 255
Leu Asp Glu Lys Asn Glu Glu Ile Ile Ser Leu Lys Asn Glu Ile Asn
260 265 270
Asp Ile Leu Asn Asp Glu Lys Ile Asp Asn Ile Ala Glu Lys Leu Lys
275 280 285
Ile Ala Lys Phe Thr Ile Ser Lys Lys Ile Asn Glu Glu Ile Lys Lys
290 295 300
Lys Asn Asp Ile Phe Asn Asn Leu Ala Asn Asp Ile Tyr Gln Phe Ile
305 310 315 320
Gly Asn Glu Tyr Tyr Phe Thr Ser Asp Ile Lys Asp Met Ile Glu Asp
325 330 335
Arg Tyr Asn Glu Leu Asn Lys Thr Ser Gln Ser Asp Leu Tyr Tyr Ile
340 345 350
Tyr Leu Leu Asn Val Phe Asp Ile Glu Lys Ile Tyr Gly Ile Tyr Leu
355 360 365
Ser Ser Tyr Gln Glu Arg Leu Gln Gln Ile Leu Gln Thr Asn Met Thr
370 375 380
Asn Leu Asp Tyr Val Glu Lys Lys Ile Gly Ser Leu Arg Asn Ile Tyr
385 390 395 400
Met Phe Leu Tyr Lys Lys Ser Lys Lys Tyr Asn Ala Leu Asp Ile Phe
405 410 415
Asp Glu Tyr Tyr Asp Tyr Val Leu Asn Tyr Asn Asp Phe Ala Lys Asp
420 425 430
Asn Glu Ile Ile Ser Ala Asp Ile Phe Ile Lys Ser Lys Pro Asp Ile
435 440 445
Pro Gln Leu Asn Phe Glu Ile Asn Asn Glu Asn Lys Asn Val Lys Tyr
450 455 460
Lys Asp Val Thr Asp Leu Asp Glu Leu Asp Asn Leu Asn Arg Ile Asn
465 470 475 480
Arg Ile Ile Asn Ile Arg Asn Val Leu Val Lys Gln Leu Lys Ile Leu
485 490 495
Tyr Asn Gln Arg Asn Asn Ile Phe Ile Lys Gln Ala Met Leu Val Lys
500 505 510
Ser Tyr Cys Tyr Lys Asn Pro Leu Asp Ile Thr Asp Phe Ser Ser Ile
515 520 525
Phe Lys Asn Asn Tyr Asn Lys Leu Lys Tyr Asp Ala Tyr Lys Glu Gly
530 535 540
Asn Gly His Ile Asn Val Ala Asp Lys Ile Asn Pro Asn Phe Val Val
545 550 555 560
Lys Tyr Leu Asn Asn Leu Tyr Lys Gln His Val Asn Lys Asn Tyr Ile
565 570 575
Leu Asn Ser Trp Asp Pro Lys Tyr Asn Arg Met Asn Lys Lys Ile Lys
580 585 590
Ile Ile Leu Ile Leu Gly Tyr Gly Gln Val Ile Gln Ile Glu Lys Gln
595 600 605
Ile Asn Arg His Ile Gly Lys Tyr Asn Ala Leu Leu Glu Lys Ala Thr
610 615 620
Leu Tyr Asn Val Gly Asn Leu Phe Thr Gln Thr Thr Asn Ile Leu Asn
625 630 635 640
Asp Ile Ser Gly Ser Leu Asn Asp Gly Leu Asp Pro Asn Ile His Asp
645 650 655
Gln Glu Asp Val Thr Val Val Glu Ser Ser Glu Ser Asn Lys Leu Ala
660 665 670
Glu Pro Glu Glu Pro Ile Glu Lys Val Glu Val Asp Arg Val Glu Lys
675 680 685
Ser Asp Asp Ala Asn Asn Ala Thr Gln Gln Val Thr Gly Thr Asp Glu
690 695 700
Ala Asn Tyr Asp Thr Ala Ser Gly Asn Ser Thr Asn Ile Asn Ile Asp
705 710 715 720
Asn Ile Asp Tyr Gly Val Asp Glu Ser Ile Arg Ile Ile Lys Tyr Ser
725 730 735
Lys Ala Glu Glu Asp Glu Tyr Asn Glu Ser Gly Asn Asn Glu Asn Glu
740 745 750
Asn Asn Glu Asn Asn Glu Asn Glu Asn Asn Glu Asn Glu Asn Asn Glu
755 760 765
Asn Glu Asn Asn Glu Asn Glu Asn Asn Glu Asn Glu Asn Ile Glu Leu
770 775 780
Lys Asn Ile Glu His Glu Asn Lys Ser Asn Ala Ser Ser Ala Ser Leu
785 790 795 800
Ser Asn Ile Phe Phe Thr Phe Ile Ile Ala Ala Leu Phe Ile Arg Pro
805 810 815
Phe Leu
<210> 60
<211> 2919
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> PfP113 人 CO
<400> 60
gccaccatgg ctaagatccc gttctttatt ctgcacatcc tcctcctgca gtttctgctt 60
tgtctgatac gctgttatgt gcacaatgat gtgattaagt tcggtgagga gaatagcctg 120
aagtgctctc agggaaactt gtatgtgttg cactgtgagg tgcagtgcct taatggcaat 180
aatgagatta tccacaagag gtgtaatgac gacattgaga aaaagtgcaa cggcaataat 240
aaatgcatat acttctttga gtacgaactg cggaagaaaa cacaaagctt tcgaaataag 300
aattctatcg agatttccga gtgtgtcgaa agcgagcaga acgaagtgaa aacctcaact 360
acctgtctcc tgagcaattc ctttattctt gacgaggcct ttatacagta tttcttcttc 420
ataaagaaca agaacgaaga gcctgtgatt tgtaaggatg ggaatatcaa cattaagagt 480
gcactcctgc actctccgtt ctgtgaaatc aaactcaagg acatttccga atatatacgc 540
aaaaagtgtg acaacaacaa ggaatgcctt atagatcctc tcgatgttca gaagaatttg 600
ctgaacgaag aagatccctg ctacatcaat aacagttacg tatctgtgaa tgtggtctgc 660
aacaaagagg aggagatagg ggatgagagc actgacagct catcaatgga gatccaggac 720
tcaacatcaa atgagcaaga cgagaatgtt aaaggaatgt caagcagcca agagatgaac 780
tcaaacaacg atgaaaacaa aaaccaagac aacgaaagcg acgatgacgt caataataat 840
aacaacaata acaatgatga ccaggacgag caaggaaacg atggcgatgt caccagctct 900
atgaacaaga atgaggacaa caaggatttg gagcatggtt cctccaatga tgtcaataac 960
aacactgaca ccttggttaa caacaaagag aataaggagt tcgtccttaa agagaagtct 1020
agccttactt ctaaaattaa caaagagctg gctcatagaa ctgccctgtt taacaaactt 1080
gcagacaaca tatcacttct gcttaacaag aaatacgatt ccttcgaaat taaggatgtg 1140
ctggaagata ggtacaacga gatgaagagg gacgcaaacc ccgatgtcta ctacatatac 1200
ctgatggata ctctggatat tgaaaagatc gaagatatca acctggaaga ggttaagatg 1260
tctctgctgg catcactgaa agaaacgatg aacaaaattg atacgatcga aaagaaaatc 1320
gaagaattta agaacaagta catctccttg tataacaagg tgaaaaccac aatgcccgaa 1380
ctctttgacc tgaatgagga tcttgtactg ctctacaacg attttccctt tgacaacggc 1440
atgatcagct ccgacatctt ctttaagtac aatccttccg agaacatcat ggatcatcag 1500
gaaatggtga agaaagggag tatcaccgaa gatgaactca ggattgttaa cgatcttgag 1560
ccactggata actatagacg tcgtaaacga attacagagc tgagaaagat tctggtggag 1620
aagctgcgga ttctgtacct ggagaagaac aatctcttca atacacaggc gagttgcatc 1680
aagagctatt gctataagaa tcctctgaac ctcaaaacct tggaagtgct cttgaagaag 1740
aactactata gactgaagga gaataaagat tacgatgttg tatccagcat tatccagcat 1800
ctcgacaatg tagacgccaa caagaagaag aaatggctga cccatgaacg gatactgaag 1860
aagctccaag ttctgattgc cgaaggctat aagcggatca acgagaaaga aaaggacatc 1920
gaccgaagaa tggctgtcta caatgccctc tatgagaaag cacagtctta caacctgcag 1980
aagcttttca acgactccaa cgattttctg aagaaatatg ccataatggg aaacagtttc 2040
gacgacggcg atgaggtttt cggttcccaa agctcaaact ttaacatctt cgatagcaat 2100
aacaccgacc agaacaatga gcaagagcag ccaaagcaag atgaccagct tttgaacaat 2160
aataacgatg acgtgctctc agagtcaaac aatgagaata aagagaaaac aagtgatgac 2220
gctactcata aggagactca ggagaaaagc gaccaggaac cttcccagaa cattcaggag 2280
gacaactccg atgagaaaca tgccgaaaac gaggagaacg tagaacagat cgaaactgat 2340
agtaatgtca gcgaagaagc caatgacgag aataaggata acatgcagac aaccactgac 2400
gaaggaaccg aagaacttca gcagaatgac gaagatgcgg agagtctgac caaggagaat 2460
tccaaatctg aggagcagga gaatgaagat tctactgacg ccgaggcgat tgataaagag 2520
gaagttgaaa cggaagagaa gggaaaggac gaacagaaga aagatgagca gaaggagcag 2580
gatgaggaag aggatggaga gaaagaaaat aagcacaaga gctccgaaac taccaatgag 2640
actgtgaccg acatcgagga aaataagaac gaggtcaaag gtgaggagca tcttcagggg 2700
tctgagcaga gcattgaggc ttccgaatca tcccagaaag atgagactaa agaaacagag 2760
gacaaggagg aatacgtgaa cgctaatgat gacgaatctt ctgaggagga cacgacgcct 2820
aacgagacaa ataaaaccga caacgggtca agtttcttct tcgcgatgag caatgcactt 2880
ctcgtgatct tgctgttgct tttcatcgaa ttcctgtaa 2919
<210> 61
<211> 970
<212> PRT
<213> 恶性疟原虫（Plasmodium falciparum）
<220>
<221> misc_feature
<223> PfP113
<400> 61
Met Ala Lys Ile Pro Phe Phe Ile Leu His Ile Leu Leu Leu Gln Phe
1 5 10 15
Leu Leu Cys Leu Ile Arg Cys Tyr Val His Asn Asp Val Ile Lys Phe
20 25 30
Gly Glu Glu Asn Ser Leu Lys Cys Ser Gln Gly Asn Leu Tyr Val Leu
35 40 45
His Cys Glu Val Gln Cys Leu Asn Gly Asn Asn Glu Ile Ile His Lys
50 55 60
Arg Cys Asn Asp Asp Ile Glu Lys Lys Cys Asn Gly Asn Asn Lys Cys
65 70 75 80
Ile Tyr Phe Phe Glu Tyr Glu Leu Arg Lys Lys Thr Gln Ser Phe Arg
85 90 95
Asn Lys Asn Ser Ile Glu Ile Ser Glu Cys Val Glu Ser Glu Gln Asn
100 105 110
Glu Val Lys Thr Ser Thr Thr Cys Leu Leu Ser Asn Ser Phe Ile Leu
115 120 125
Asp Glu Ala Phe Ile Gln Tyr Phe Phe Phe Ile Lys Asn Lys Asn Glu
130 135 140
Glu Pro Val Ile Cys Lys Asp Gly Asn Ile Asn Ile Lys Ser Ala Leu
145 150 155 160
Leu His Ser Pro Phe Cys Glu Ile Lys Leu Lys Asp Ile Ser Glu Tyr
165 170 175
Ile Arg Lys Lys Cys Asp Asn Asn Lys Glu Cys Leu Ile Asp Pro Leu
180 185 190
Asp Val Gln Lys Asn Leu Leu Asn Glu Glu Asp Pro Cys Tyr Ile Asn
195 200 205
Asn Ser Tyr Val Ser Val Asn Val Val Cys Asn Lys Glu Glu Glu Ile
210 215 220
Gly Asp Glu Ser Thr Asp Ser Ser Ser Met Glu Ile Gln Asp Ser Thr
225 230 235 240
Ser Asn Glu Gln Asp Glu Asn Val Lys Gly Met Ser Ser Ser Gln Glu
245 250 255
Met Asn Ser Asn Asn Asp Glu Asn Lys Asn Gln Asp Asn Glu Ser Asp
260 265 270
Asp Asp Val Asn Asn Asn Asn Asn Asn Asn Asn Asp Asp Gln Asp Glu
275 280 285
Gln Gly Asn Asp Gly Asp Val Thr Ser Ser Met Asn Lys Asn Glu Asp
290 295 300
Asn Lys Asp Leu Glu His Gly Ser Ser Asn Asp Val Asn Asn Asn Thr
305 310 315 320
Asp Thr Leu Val Asn Asn Lys Glu Asn Lys Glu Phe Val Leu Lys Glu
325 330 335
Lys Ser Ser Leu Thr Ser Lys Ile Asn Lys Glu Leu Ala His Arg Thr
340 345 350
Ala Leu Phe Asn Lys Leu Ala Asp Asn Ile Ser Leu Leu Leu Asn Lys
355 360 365
Lys Tyr Asp Ser Phe Glu Ile Lys Asp Val Leu Glu Asp Arg Tyr Asn
370 375 380
Glu Met Lys Arg Asp Ala Asn Pro Asp Val Tyr Tyr Ile Tyr Leu Met
385 390 395 400
Asp Thr Leu Asp Ile Glu Lys Ile Glu Asp Ile Asn Leu Glu Glu Val
405 410 415
Lys Met Ser Leu Leu Ala Ser Leu Lys Glu Thr Met Asn Lys Ile Asp
420 425 430
Thr Ile Glu Lys Lys Ile Glu Glu Phe Lys Asn Lys Tyr Ile Ser Leu
435 440 445
Tyr Asn Lys Val Lys Thr Thr Met Pro Glu Leu Phe Asp Leu Asn Glu
450 455 460
Asp Leu Val Leu Leu Tyr Asn Asp Phe Pro Phe Asp Asn Gly Met Ile
465 470 475 480
Ser Ser Asp Ile Phe Phe Lys Tyr Asn Pro Ser Glu Asn Ile Met Asp
485 490 495
His Gln Glu Met Val Lys Lys Gly Ser Ile Thr Glu Asp Glu Leu Arg
500 505 510
Ile Val Asn Asp Leu Glu Pro Leu Asp Asn Tyr Arg Arg Arg Lys Arg
515 520 525
Ile Thr Glu Leu Arg Lys Ile Leu Val Glu Lys Leu Arg Ile Leu Tyr
530 535 540
Leu Glu Lys Asn Asn Leu Phe Asn Thr Gln Ala Ser Cys Ile Lys Ser
545 550 555 560
Tyr Cys Tyr Lys Asn Pro Leu Asn Leu Lys Thr Leu Glu Val Leu Leu
565 570 575
Lys Lys Asn Tyr Tyr Arg Leu Lys Glu Asn Lys Asp Tyr Asp Val Val
580 585 590
Ser Ser Ile Ile Gln His Leu Asp Asn Val Asp Ala Asn Lys Lys Lys
595 600 605
Lys Trp Leu Thr His Glu Arg Ile Leu Lys Lys Leu Gln Val Leu Ile
610 615 620
Ala Glu Gly Tyr Lys Arg Ile Asn Glu Lys Glu Lys Asp Ile Asp Arg
625 630 635 640
Arg Met Ala Val Tyr Asn Ala Leu Tyr Glu Lys Ala Gln Ser Tyr Asn
645 650 655
Leu Gln Lys Leu Phe Asn Asp Ser Asn Asp Phe Leu Lys Lys Tyr Ala
660 665 670
Ile Met Gly Asn Ser Phe Asp Asp Gly Asp Glu Val Phe Gly Ser Gln
675 680 685
Ser Ser Asn Phe Asn Ile Phe Asp Ser Asn Asn Thr Asp Gln Asn Asn
690 695 700
Glu Gln Glu Gln Pro Lys Gln Asp Asp Gln Leu Leu Asn Asn Asn Asn
705 710 715 720
Asp Asp Val Leu Ser Glu Ser Asn Asn Glu Asn Lys Glu Lys Thr Ser
725 730 735
Asp Asp Ala Thr His Lys Glu Thr Gln Glu Lys Ser Asp Gln Glu Pro
740 745 750
Ser Gln Asn Ile Gln Glu Asp Asn Ser Asp Glu Lys His Ala Glu Asn
755 760 765
Glu Glu Asn Val Glu Gln Ile Glu Thr Asp Ser Asn Val Ser Glu Glu
770 775 780
Ala Asn Asp Glu Asn Lys Asp Asn Met Gln Thr Thr Thr Asp Glu Gly
785 790 795 800
Thr Glu Glu Leu Gln Gln Asn Asp Glu Asp Ala Glu Ser Leu Thr Lys
805 810 815
Glu Asn Ser Lys Ser Glu Glu Gln Glu Asn Glu Asp Ser Thr Asp Ala
820 825 830
Glu Ala Ile Asp Lys Glu Glu Val Glu Thr Glu Glu Lys Gly Lys Asp
835 840 845
Glu Gln Lys Lys Asp Glu Gln Lys Glu Gln Asp Glu Glu Glu Asp Gly
850 855 860
Glu Lys Glu Asn Lys His Lys Ser Ser Glu Thr Thr Asn Glu Thr Val
865 870 875 880
Thr Asp Ile Glu Glu Asn Lys Asn Glu Val Lys Gly Glu Glu His Leu
885 890 895
Gln Gly Ser Glu Gln Ser Ile Glu Ala Ser Glu Ser Ser Gln Lys Asp
900 905 910
Glu Thr Lys Glu Thr Glu Asp Lys Glu Glu Tyr Val Asn Ala Asn Asp
915 920 925
Asp Glu Ser Ser Glu Glu Asp Thr Thr Pro Asn Glu Thr Asn Lys Thr
930 935 940
Asp Asn Gly Ser Ser Phe Phe Phe Ala Met Ser Asn Ala Leu Leu Val
945 950 955 960
Ile Leu Leu Leu Leu Phe Ile Glu Phe Leu
965 970
<210> 62
<211> 969
<212> PRT
<213> 恶性疟原虫（Plasmodium falciparum）
<220>
<221> misc_feature
<223> PfP113
<400> 62
Met Lys Ile Pro Phe Phe Ile Leu His Ile Leu Leu Leu Gln Phe Leu
1 5 10 15
Leu Cys Leu Ile Arg Cys Tyr Val His Asn Asp Val Ile Lys Phe Gly
20 25 30
Glu Glu Asn Ser Leu Lys Cys Ser Gln Gly Asn Leu Tyr Val Leu His
35 40 45
Cys Glu Val Gln Cys Leu Asn Gly Asn Asn Glu Ile Ile His Lys Arg
50 55 60
Cys Asn Asp Asp Ile Glu Lys Lys Cys Asn Gly Asn Asn Lys Cys Ile
65 70 75 80
Tyr Phe Phe Glu Tyr Glu Leu Arg Lys Lys Thr Gln Ser Phe Arg Asn
85 90 95
Lys Asn Ser Ile Glu Ile Ser Glu Cys Val Glu Ser Glu Gln Asn Glu
100 105 110
Val Lys Thr Ser Thr Thr Cys Leu Leu Ser Asn Ser Phe Ile Leu Asp
115 120 125
Glu Ala Phe Ile Gln Tyr Phe Phe Phe Ile Lys Asn Lys Asn Glu Glu
130 135 140
Pro Val Ile Cys Lys Asp Gly Asn Ile Asn Ile Lys Ser Ala Leu Leu
145 150 155 160
His Ser Pro Phe Cys Glu Ile Lys Leu Lys Asp Ile Ser Glu Tyr Ile
165 170 175
Arg Lys Lys Cys Asp Asn Asn Lys Glu Cys Leu Ile Asp Pro Leu Asp
180 185 190
Val Gln Lys Asn Leu Leu Asn Glu Glu Asp Pro Cys Tyr Ile Asn Asn
195 200 205
Ser Tyr Val Ser Val Asn Val Val Cys Asn Lys Glu Glu Glu Ile Gly
210 215 220
Asp Glu Ser Thr Asp Ser Ser Ser Met Glu Ile Gln Asp Ser Thr Ser
225 230 235 240
Asn Glu Gln Asp Glu Asn Val Lys Gly Met Ser Ser Ser Gln Glu Met
245 250 255
Asn Ser Asn Asn Asp Glu Asn Lys Asn Gln Asp Asn Glu Ser Asp Asp
260 265 270
Asp Val Asn Asn Asn Asn Asn Asn Asn Asn Asp Asp Gln Asp Glu Gln
275 280 285
Gly Asn Asp Gly Asp Val Thr Ser Ser Met Asn Lys Asn Glu Asp Asn
290 295 300
Lys Asp Leu Glu His Gly Ser Ser Asn Asp Val Asn Asn Asn Thr Asp
305 310 315 320
Thr Leu Val Asn Asn Lys Glu Asn Lys Glu Phe Val Leu Lys Glu Lys
325 330 335
Ser Ser Leu Thr Ser Lys Ile Asn Lys Glu Leu Ala His Arg Thr Ala
340 345 350
Leu Phe Asn Lys Leu Ala Asp Asn Ile Ser Leu Leu Leu Asn Lys Lys
355 360 365
Tyr Asp Ser Phe Glu Ile Lys Asp Val Leu Glu Asp Arg Tyr Asn Glu
370 375 380
Met Lys Arg Asp Ala Asn Pro Asp Val Tyr Tyr Ile Tyr Leu Met Asp
385 390 395 400
Thr Leu Asp Ile Glu Lys Ile Glu Asp Ile Asn Leu Glu Glu Val Lys
405 410 415
Met Ser Leu Leu Ala Ser Leu Lys Glu Thr Met Asn Lys Ile Asp Thr
420 425 430
Ile Glu Lys Lys Ile Glu Glu Phe Lys Asn Lys Tyr Ile Ser Leu Tyr
435 440 445
Asn Lys Val Lys Thr Thr Met Pro Glu Leu Phe Asp Leu Asn Glu Asp
450 455 460
Leu Val Leu Leu Tyr Asn Asp Phe Pro Phe Asp Asn Gly Met Ile Ser
465 470 475 480
Ser Asp Ile Phe Phe Lys Tyr Asn Pro Ser Glu Asn Ile Met Asp His
485 490 495
Gln Glu Met Val Lys Lys Gly Ser Ile Thr Glu Asp Glu Leu Arg Ile
500 505 510
Val Asn Asp Leu Glu Pro Leu Asp Asn Tyr Arg Arg Arg Lys Arg Ile
515 520 525
Thr Glu Leu Arg Lys Ile Leu Val Glu Lys Leu Arg Ile Leu Tyr Leu
530 535 540
Glu Lys Asn Asn Leu Phe Asn Thr Gln Ala Ser Cys Ile Lys Ser Tyr
545 550 555 560
Cys Tyr Lys Asn Pro Leu Asn Leu Lys Thr Leu Glu Val Leu Leu Lys
565 570 575
Lys Asn Tyr Tyr Arg Leu Lys Glu Asn Lys Asp Tyr Asp Val Val Ser
580 585 590
Ser Ile Ile Gln His Leu Asp Asn Val Asp Ala Asn Lys Lys Lys Lys
595 600 605
Trp Leu Thr His Glu Arg Ile Leu Lys Lys Leu Gln Val Leu Ile Ala
610 615 620
Glu Gly Tyr Lys Arg Ile Asn Glu Lys Glu Lys Asp Ile Asp Arg Arg
625 630 635 640
Met Ala Val Tyr Asn Ala Leu Tyr Glu Lys Ala Gln Ser Tyr Asn Leu
645 650 655
Gln Lys Leu Phe Asn Asp Ser Asn Asp Phe Leu Lys Lys Tyr Ala Ile
660 665 670
Met Gly Asn Ser Phe Asp Asp Gly Asp Glu Val Phe Gly Ser Gln Ser
675 680 685
Ser Asn Phe Asn Ile Phe Asp Ser Asn Asn Thr Asp Gln Asn Asn Glu
690 695 700
Gln Glu Gln Pro Lys Gln Asp Asp Gln Leu Leu Asn Asn Asn Asn Asp
705 710 715 720
Asp Val Leu Ser Glu Ser Asn Asn Glu Asn Lys Glu Lys Thr Ser Asp
725 730 735
Asp Ala Thr His Lys Glu Thr Gln Glu Lys Ser Asp Gln Glu Pro Ser
740 745 750
Gln Asn Ile Gln Glu Asp Asn Ser Asp Glu Lys His Ala Glu Asn Glu
755 760 765
Glu Asn Val Glu Gln Ile Glu Thr Asp Ser Asn Val Ser Glu Glu Ala
770 775 780
Asn Asp Glu Asn Lys Asp Asn Met Gln Thr Thr Thr Asp Glu Gly Thr
785 790 795 800
Glu Glu Leu Gln Gln Asn Asp Glu Asp Ala Glu Ser Leu Thr Lys Glu
805 810 815
Asn Ser Lys Ser Glu Glu Gln Glu Asn Glu Asp Ser Thr Asp Ala Glu
820 825 830
Ala Ile Asp Lys Glu Glu Val Glu Thr Glu Glu Lys Gly Lys Asp Glu
835 840 845
Gln Lys Lys Asp Glu Gln Lys Glu Gln Asp Glu Glu Glu Asp Gly Glu
850 855 860
Lys Glu Asn Lys His Lys Ser Ser Glu Thr Thr Asn Glu Thr Val Thr
865 870 875 880
Asp Ile Glu Glu Asn Lys Asn Glu Val Lys Gly Glu Glu His Leu Gln
885 890 895
Gly Ser Glu Gln Ser Ile Glu Ala Ser Glu Ser Ser Gln Lys Asp Glu
900 905 910
Thr Lys Glu Thr Glu Asp Lys Glu Glu Tyr Val Asn Ala Asn Asp Asp
915 920 925
Glu Ser Ser Glu Glu Asp Thr Thr Pro Asn Glu Thr Asn Lys Thr Asp
930 935 940
Asn Gly Ser Ser Phe Phe Phe Ala Met Ser Asn Ala Leu Leu Val Ile
945 950 955 960
Leu Leu Leu Leu Phe Ile Glu Phe Leu
965
<210> 63
<211> 3141
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> PvP113 人 CO
<400> 63
gccaccatgg ctaagctccc gcccctctgc aggttgccac ttgcgctggt gcttctttgt 60
ctgacatcca gagcgcgctg ttatgtacac aacgacgtga tgaaatttgg tgaagaaaac 120
tctctcaagt gttcacaagg gtctctctac atccttcact gcgaggtcaa gtgtgtgaat 180
gcaaagaaca gaatcatcca ccgaagttgc atcgaccagg ttgaggcgaa gtgcatgggc 240
aacgccaaat gtaagtacta ttttgactat gtcgtgaaat cccgcggaca gagtttgcgg 300
aacaaaaatg aaatcgaaat agaagaatgc gtggaatccg aacggaatga gattaagacg 360
tccacaactt gccttctgtc aaattcattt ttgctggatg agacctacat tcaatatttc 420
tttttcatca aaaacaaaaa tgaagaaccc ataacgtgtc gagacggtag actgagcgta 480
aagagtgcga tacttcatag tcccttttgc aaaataaatc ttaaggacat cactgagatt 540
ctcaagagac aatgcgatca cagtaaggag tgcgtcatta atccgtatgt cctccaaaaa 600
gatgcattga atgagagaga ccagtgttac attaacaact catatgtctc actcaatgtc 660
gtttgcacca aagaagggga ggagcaacca gaagagagtg gacataaaca gaagagggat 720
gacgacgtgg acgaaaccga ggaaggctca tatgacgtct ctgcggatca gaacaaatca 780
gcaatagtcg gcgagggaaa cgatgatccg gaatccctgg gtgaagaaga tgagctttca 840
gagactaacg aggcagttga tctcattatg aactccaagg aaagctttga aaacaaaatt 900
cggaaggcga agtcaatact tctctcacag atgaacgagc aggaggtgaa aaagaatgca 960
atattcaaga aactcgggga agagctttcc aaaatggtcc tccagaaata tgaaccaagt 1020
gatctgaaag acttgatcga ggacaggtat aatgaaatga gacggtcccc tgaccaagac 1080
ctttattatc tctaccttat agatacactc gaaataaata aaatggaaga tcttgacgtg 1140
actgcactgc aagaccagct cgctatattg ttggaagagc agatgggcaa gatgaatcgc 1200
attgagaaaa cgattaaccg attgaggaaa aaataccttt ctatttacaa caaagcgaag 1260
aacaagaaag ttaaagacat ttacgatgaa ggagttgatc ctgtactcac gtacgacgac 1320
ttcgcgcacg gcaacggcat tattacggca gatattttct ttaagtataa gcctgccatt 1380
aagccgttga cttttagcaa atctaatgcc tctgaggagc gggggtcatc caaaaaaaaa 1440
gagtacaagg accttcttga gatggacgcg ctggatgagt acaaccggaa gaaacggata 1500
actgacatgc gaaacggtct gatggagact ttgaagaaaa tgtattacgc aaaaaatggg 1560
atattcaata atctggcgag ttgcattaag tcatattgct ataaaaggcc tctcaacctt 1620
aatgcacttt cctccgttct taaaagaaat tttgaaaatc ttcgagagaa aaaatcaacc 1680
gatccagtgg cccccatagt cagatatttg caaaaggtca gtggcgaggt tggcggggaa 1740
gtgggaggcg cggctggcgg tgctgcgggg ggtgctgcaa gtggtgccgc atcaggagct 1800
gtcagtgaag cagtaggagg cgccataggt ggggctgtgg gcgaggcatc tatcgcagta 1860
aatccgcccc gatgggaaaa atcacgacga attcttcaaa agctcaaggc tttgctgcac 1920
ctgggttatc aacaggcact ggacaaagag ttggagatcg acgagaggac tgacaaatac 1980
aacgctctga atgagaaagc aaaagaatat aaccttcagc ggcttttttc cgagtcagac 2040
aagttgctta agaaggtcgc aatgcttacg tcagccagtg aaagtgcaga tgaagtattc 2100
ggtaaccaag cgagcttttt cgatgtttac agaggagagg cagcgtcaca aaaaggagta 2160
gccgccagtg aaaagggcgt agctgcaaga gagaaaggag tcgccgcgag tgagaagggg 2220
gtcgcggcat cagaaaaggg tgtggcggcc agcgagaagg gtgtcgcggc atcagaaaaa 2280
ggtgttgccg caagccaaaa aggcggagaa acgtctgagg aaggggaggc tgccagtcaa 2340
aaggatgttg ctacgtccga ggagggtgga gctagcagcg agaaacgcga tgacgacgag 2400
atgaatccgc ccgacgaagg gtatgagtcc gctaaggaag acggggagaa tgcacaagat 2460
gacgacagcg gcggaaatgc tgaacccgta gagggaaaag caggcgaaag tgaagacgct 2520
gatggggtaa atgccggctc caataaagaa ggtgaggatg gggagagcgt tgaagaagaa 2580
gctgcggaag gggaagcggc acccaaggag gaggctgcag acggagaaga tgctccggag 2640
gaggaagccg agggtgagga tgccccagaa gaagaggccg atggcgaaga cgccccgaag 2700
gaggaggcta ctgacggaga ggacgcccct aaagaggaag aggccgaagg tgaggatgca 2760
agcaaggacg aggaggccga cgaaggctca actgacgagg aagaagccgc cgacggcggg 2820
agtactgatg ctacagcagc cgatgaagct gcaggcggag tcgcggacca aaacgacgtg 2880
cccgttaagg gagaagactc agatggcgcg gaatctgatg gagcagaaga tgccgccact 2940
gaaattaggg gtgaggctga agccggtgaa gaggccgcgg agcaacctac tggagaggcc 3000
gtagtcaaag gggattccga gggcggggct tctgggctgg aaaccgagaa gaagggagac 3060
gacgggggct ccttcttcca aggtctttcc cgagtgttgc ttactgttct tgcaatactt 3120
tccttggaat ttctcctttg a 3141
<210> 64
<211> 1044
<212> PRT
<213> 间日疟原虫（Plasmodium vivax）
<220>
<221> misc_feature
<223> PvP113
<400> 64
Met Ala Lys Leu Pro Pro Leu Cys Arg Leu Pro Leu Ala Leu Val Leu
1 5 10 15
Leu Cys Leu Thr Ser Arg Ala Arg Cys Tyr Val His Asn Asp Val Met
20 25 30
Lys Phe Gly Glu Glu Asn Ser Leu Lys Cys Ser Gln Gly Ser Leu Tyr
35 40 45
Ile Leu His Cys Glu Val Lys Cys Val Asn Ala Lys Asn Arg Ile Ile
50 55 60
His Arg Ser Cys Ile Asp Gln Val Glu Ala Lys Cys Met Gly Asn Ala
65 70 75 80
Lys Cys Lys Tyr Tyr Phe Asp Tyr Val Val Lys Ser Arg Gly Gln Ser
85 90 95
Leu Arg Asn Lys Asn Glu Ile Glu Ile Glu Glu Cys Val Glu Ser Glu
100 105 110
Arg Asn Glu Ile Lys Thr Ser Thr Thr Cys Leu Leu Ser Asn Ser Phe
115 120 125
Leu Leu Asp Glu Thr Tyr Ile Gln Tyr Phe Phe Phe Ile Lys Asn Lys
130 135 140
Asn Glu Glu Pro Ile Thr Cys Arg Asp Gly Arg Leu Ser Val Lys Ser
145 150 155 160
Ala Ile Leu His Ser Pro Phe Cys Lys Ile Asn Leu Lys Asp Ile Thr
165 170 175
Glu Ile Leu Lys Arg Gln Cys Asp His Ser Lys Glu Cys Val Ile Asn
180 185 190
Pro Tyr Val Leu Gln Lys Asp Ala Leu Asn Glu Arg Asp Gln Cys Tyr
195 200 205
Ile Asn Asn Ser Tyr Val Ser Leu Asn Val Val Cys Thr Lys Glu Gly
210 215 220
Glu Glu Gln Pro Glu Glu Ser Gly His Lys Gln Lys Arg Asp Asp Asp
225 230 235 240
Val Asp Glu Thr Glu Glu Gly Ser Tyr Asp Val Ser Ala Asp Gln Asn
245 250 255
Lys Ser Ala Ile Val Gly Glu Gly Asn Asp Asp Pro Glu Ser Leu Gly
260 265 270
Glu Glu Asp Glu Leu Ser Glu Thr Asn Glu Ala Val Asp Leu Ile Met
275 280 285
Asn Ser Lys Glu Ser Phe Glu Asn Lys Ile Arg Lys Ala Lys Ser Ile
290 295 300
Leu Leu Ser Gln Met Asn Glu Gln Glu Val Lys Lys Asn Ala Ile Phe
305 310 315 320
Lys Lys Leu Gly Glu Glu Leu Ser Lys Met Val Leu Gln Lys Tyr Glu
325 330 335
Pro Ser Asp Leu Lys Asp Leu Ile Glu Asp Arg Tyr Asn Glu Met Arg
340 345 350
Arg Ser Pro Asp Gln Asp Leu Tyr Tyr Leu Tyr Leu Ile Asp Thr Leu
355 360 365
Glu Ile Asn Lys Met Glu Asp Leu Asp Val Thr Ala Leu Gln Asp Gln
370 375 380
Leu Ala Ile Leu Leu Glu Glu Gln Met Gly Lys Met Asn Arg Ile Glu
385 390 395 400
Lys Thr Ile Asn Arg Leu Arg Lys Lys Tyr Leu Ser Ile Tyr Asn Lys
405 410 415
Ala Lys Asn Lys Lys Val Lys Asp Ile Tyr Asp Glu Gly Val Asp Pro
420 425 430
Val Leu Thr Tyr Asp Asp Phe Ala His Gly Asn Gly Ile Ile Thr Ala
435 440 445
Asp Ile Phe Phe Lys Tyr Lys Pro Ala Ile Lys Pro Leu Thr Phe Ser
450 455 460
Lys Ser Asn Ala Ser Glu Glu Arg Gly Ser Ser Lys Lys Lys Glu Tyr
465 470 475 480
Lys Asp Leu Leu Glu Met Asp Ala Leu Asp Glu Tyr Asn Arg Lys Lys
485 490 495
Arg Ile Thr Asp Met Arg Asn Gly Leu Met Glu Thr Leu Lys Lys Met
500 505 510
Tyr Tyr Ala Lys Asn Gly Ile Phe Asn Asn Leu Ala Ser Cys Ile Lys
515 520 525
Ser Tyr Cys Tyr Lys Arg Pro Leu Asn Leu Asn Ala Leu Ser Ser Val
530 535 540
Leu Lys Arg Asn Phe Glu Asn Leu Arg Glu Lys Lys Ser Thr Asp Pro
545 550 555 560
Val Ala Pro Ile Val Arg Tyr Leu Gln Lys Val Ser Gly Glu Val Gly
565 570 575
Gly Glu Val Gly Gly Ala Ala Gly Gly Ala Ala Gly Gly Ala Ala Ser
580 585 590
Gly Ala Ala Ser Gly Ala Val Ser Glu Ala Val Gly Gly Ala Ile Gly
595 600 605
Gly Ala Val Gly Glu Ala Ser Ile Ala Val Asn Pro Pro Arg Trp Glu
610 615 620
Lys Ser Arg Arg Ile Leu Gln Lys Leu Lys Ala Leu Leu His Leu Gly
625 630 635 640
Tyr Gln Gln Ala Leu Asp Lys Glu Leu Glu Ile Asp Glu Arg Thr Asp
645 650 655
Lys Tyr Asn Ala Leu Asn Glu Lys Ala Lys Glu Tyr Asn Leu Gln Arg
660 665 670
Leu Phe Ser Glu Ser Asp Lys Leu Leu Lys Lys Val Ala Met Leu Thr
675 680 685
Ser Ala Ser Glu Ser Ala Asp Glu Val Phe Gly Asn Gln Ala Ser Phe
690 695 700
Phe Asp Val Tyr Arg Gly Glu Ala Ala Ser Gln Lys Gly Val Ala Ala
705 710 715 720
Ser Glu Lys Gly Val Ala Ala Arg Glu Lys Gly Val Ala Ala Ser Glu
725 730 735
Lys Gly Val Ala Ala Ser Glu Lys Gly Val Ala Ala Ser Glu Lys Gly
740 745 750
Val Ala Ala Ser Glu Lys Gly Val Ala Ala Ser Gln Lys Gly Gly Glu
755 760 765
Thr Ser Glu Glu Gly Glu Ala Ala Ser Gln Lys Asp Val Ala Thr Ser
770 775 780
Glu Glu Gly Gly Ala Ser Ser Glu Lys Arg Asp Asp Asp Glu Met Asn
785 790 795 800
Pro Pro Asp Glu Gly Tyr Glu Ser Ala Lys Glu Asp Gly Glu Asn Ala
805 810 815
Gln Asp Asp Asp Ser Gly Gly Asn Ala Glu Pro Val Glu Gly Lys Ala
820 825 830
Gly Glu Ser Glu Asp Ala Asp Gly Val Asn Ala Gly Ser Asn Lys Glu
835 840 845
Gly Glu Asp Gly Glu Ser Val Glu Glu Glu Ala Ala Glu Gly Glu Ala
850 855 860
Ala Pro Lys Glu Glu Ala Ala Asp Gly Glu Asp Ala Pro Glu Glu Glu
865 870 875 880
Ala Glu Gly Glu Asp Ala Pro Glu Glu Glu Ala Asp Gly Glu Asp Ala
885 890 895
Pro Lys Glu Glu Ala Thr Asp Gly Glu Asp Ala Pro Lys Glu Glu Glu
900 905 910
Ala Glu Gly Glu Asp Ala Ser Lys Asp Glu Glu Ala Asp Glu Gly Ser
915 920 925
Thr Asp Glu Glu Glu Ala Ala Asp Gly Gly Ser Thr Asp Ala Thr Ala
930 935 940
Ala Asp Glu Ala Ala Gly Gly Val Ala Asp Gln Asn Asp Val Pro Val
945 950 955 960
Lys Gly Glu Asp Ser Asp Gly Ala Glu Ser Asp Gly Ala Glu Asp Ala
965 970 975
Ala Thr Glu Ile Arg Gly Glu Ala Glu Ala Gly Glu Glu Ala Ala Glu
980 985 990
Gln Pro Thr Gly Glu Ala Val Val Lys Gly Asp Ser Glu Gly Gly Ala
995 1000 1005
Ser Gly Leu Glu Thr Glu Lys Lys Gly Asp Asp Gly Gly Ser Phe
1010 1015 1020
Phe Gln Gly Leu Ser Arg Val Leu Leu Thr Val Leu Ala Ile Leu
1025 1030 1035
Ser Leu Glu Phe Leu Leu
1040
<210> 65
<211> 969
<212> PRT
<213> 间日疟原虫（Plasmodium vivax）
<220>
<221> misc_feature
<223> P113
<400> 65
Met Lys Leu Pro Pro Leu Cys Arg Leu Pro Leu Ala Leu Val Leu Leu
1 5 10 15
Cys Leu Thr Ser Arg Ala Arg Cys Tyr Val His Asn Asp Val Met Lys
20 25 30
Phe Gly Glu Glu Asn Ser Leu Lys Cys Ser Gln Gly Ser Leu Tyr Ile
35 40 45
Leu His Cys Glu Val Lys Cys Val Asn Ala Lys Asn Arg Ile Ile His
50 55 60
Arg Ser Cys Ile Asp Gln Val Glu Ala Lys Cys Met Gly Asn Ala Lys
65 70 75 80
Cys Lys Tyr Tyr Phe Asp Tyr Val Val Lys Ser Arg Gly Gln Ser Leu
85 90 95
Arg Asn Lys Asn Glu Ile Glu Ile Glu Glu Cys Val Glu Ser Glu Arg
100 105 110
Asn Glu Ile Lys Thr Ser Thr Thr Cys Leu Leu Ser Asn Ser Phe Leu
115 120 125
Leu Asp Glu Thr Tyr Ile Gln Tyr Phe Phe Phe Ile Lys Asn Lys Asn
130 135 140
Glu Glu Pro Ile Thr Cys Arg Asp Gly Arg Leu Ser Val Lys Ser Ala
145 150 155 160
Ile Leu His Ser Pro Phe Cys Lys Ile Asn Leu Lys Asp Ile Thr Glu
165 170 175
Ile Leu Lys Arg Gln Cys Asp His Ser Lys Glu Cys Val Ile Asn Pro
180 185 190
Tyr Val Leu Gln Lys Asp Ala Leu Asn Glu Arg Asp Gln Cys Tyr Ile
195 200 205
Asn Asn Ser Tyr Val Ser Leu Asn Val Val Cys Thr Lys Glu Gly Glu
210 215 220
Glu Gln Pro Glu Glu Ser Gly His Lys Gln Lys Arg Asp Asp Asp Val
225 230 235 240
Asp Glu Thr Glu Glu Gly Ser Tyr Asp Val Ser Ala Asp Gln Asn Lys
245 250 255
Ser Ala Ile Val Gly Glu Gly Asn Asp Asp Pro Glu Ser Leu Gly Glu
260 265 270
Glu Asp Glu Leu Ser Glu Thr Asn Glu Ala Val Asp Leu Ile Met Asn
275 280 285
Ser Lys Glu Ser Phe Glu Asn Lys Ile Arg Lys Ala Lys Ser Ile Leu
290 295 300
Leu Ser Gln Met Asn Glu Gln Glu Val Lys Lys Asn Ala Ile Phe Lys
305 310 315 320
Lys Leu Gly Glu Glu Leu Ser Lys Met Val Leu Gln Lys Tyr Glu Pro
325 330 335
Ser Asp Leu Lys Asp Leu Ile Glu Asp Arg Tyr Asn Glu Met Arg Arg
340 345 350
Ser Pro Asp Gln Asp Leu Tyr Tyr Leu Tyr Leu Ile Asp Thr Leu Glu
355 360 365
Ile Asn Lys Met Glu Asn Leu Asp Val Thr Ala Leu Gln Asp Gln Leu
370 375 380
Ala Ile Leu Leu Glu Glu Gln Met Gly Lys Met Asn Arg Ile Glu Lys
385 390 395 400
Thr Ile Asn Arg Leu Arg Lys Lys Tyr Leu Ser Ile Tyr Asn Lys Ala
405 410 415
Lys Asn Lys Lys Val Lys Asp Ile Tyr Asp Glu Gly Val Asp Pro Val
420 425 430
Leu Thr Tyr Asp Asp Phe Ala His Gly Asn Gly Ile Ile Thr Ala Asp
435 440 445
Ile Phe Phe Lys Tyr Lys Pro Ala Ile Lys Pro Leu Thr Phe Ser Lys
450 455 460
Ser Asn Ala Ser Glu Glu Arg Gly Ser Ser Lys Lys Lys Glu Tyr Lys
465 470 475 480
Asp Leu Leu Glu Met Asp Ala Leu Asp Glu Tyr Asn Arg Lys Lys Arg
485 490 495
Ile Thr Asp Met Arg Asn Gly Leu Met Glu Thr Leu Lys Lys Met Tyr
500 505 510
Tyr Ala Lys Asn Gly Ile Phe Asn Asn Leu Ala Ser Cys Ile Lys Ser
515 520 525
Tyr Cys Tyr Lys Arg Pro Leu Asn Leu Asn Ala Leu Ser Ser Val Leu
530 535 540
Lys Arg Asn Phe Glu Asn Leu Arg Glu Lys Lys Ser Thr Asp Pro Val
545 550 555 560
Ala Pro Ile Val Arg Tyr Leu Gln Lys Lys Ser Arg Arg Ile Leu Gln
565 570 575
Lys Leu Lys Ala Leu Leu His Leu Gly Tyr Gln Gln Ala Leu Asp Lys
580 585 590
Glu Leu Glu Ile Asp Glu Arg Thr Asp Lys Tyr Asn Ala Leu Asn Glu
595 600 605
Lys Ala Lys Glu Tyr Asn Leu Gln Arg Leu Phe Ser Glu Ser Asp Lys
610 615 620
Leu Leu Lys Lys Val Ala Met Leu Thr Ser Ala Ser Glu Ser Ala Asp
625 630 635 640
Glu Val Phe Gly Asn Gln Ala Ser Phe Phe Asp Val Tyr Arg Gly Glu
645 650 655
Ala Ala Ser Glu Lys Gly Val Ala Ala Ser Gln Lys Gly Gly Glu Thr
660 665 670
Ser Glu Glu Gly Glu Ala Ala Ser Gln Lys Asp Val Ala Thr Ser Glu
675 680 685
Glu Gly Gly Ala Ser Ser Glu Lys Arg Asp Asp Asp Glu Met Asn Pro
690 695 700
Pro Asp Glu Gly Tyr Glu Ser Ala Lys Glu Asp Gly Glu Asn Ala Gln
705 710 715 720
Asp Asp Asp Ser Gly Gly Asn Ala Glu Pro Val Glu Gly Lys Ala Gly
725 730 735
Glu Ser Glu Asp Ala Asp Gly Val Asn Ala Gly Ser Asn Lys Glu Gly
740 745 750
Glu Asp Gly Glu Ser Val Glu Glu Glu Ala Ala Glu Gly Glu Ala Ala
755 760 765
Pro Lys Glu Glu Ala Ala Asp Gly Glu Asp Ala Pro Glu Glu Glu Ala
770 775 780
Glu Gly Glu Asp Ala Pro Lys Glu Glu Ala Ala Asp Gly Glu Ala Ser
785 790 795 800
Ser Lys Glu Glu Glu Thr Asp Gly Glu Val Ala Pro Glu Glu Ala Ala
805 810 815
Asp Gly Glu Asp Ala Pro Lys Glu Glu Ala Thr Asp Gly Glu Asp Ala
820 825 830
Pro Lys Glu Glu Glu Ala Glu Gly Glu Asp Ala Ser Lys Asp Glu Glu
835 840 845
Ala Asp Glu Gly Ser Thr Asp Glu Glu Glu Ala Ala Asp Gly Gly Ser
850 855 860
Thr Asp Ala Thr Ala Ala Asp Glu Ala Ala Gly Gly Val Ala Asp Gln
865 870 875 880
Asn Asp Val Pro Val Lys Gly Glu Asp Ser Asp Gly Ala Glu Ser Asp
885 890 895
Gly Ala Glu Asp Ala Ala Thr Glu Ile Arg Gly Glu Ala Glu Ala Gly
900 905 910
Glu Glu Ala Ala Glu Gln Pro Thr Gly Glu Ala Val Val Lys Gly Asp
915 920 925
Ser Glu Gly Gly Ala Ser Gly Leu Glu Thr Glu Lys Lys Gly Asp Asp
930 935 940
Gly Gly Ser Phe Phe Gln Gly Leu Ser Arg Val Leu Leu Thr Val Leu
945 950 955 960
Ala Ile Leu Ser Leu Glu Phe Leu Leu
965
<210> 66
<211> 654
<212> DNA
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> Ag40
<400> 66
ggatccgcca ccatggctgg ggcgagtatg agccacttgc agtgtctgac atctgttgct 60
ggtctgtcct ctatcgtcat gtcaatgttc cccaaactca ttgccaataa tccttccctg 120
tttagaccac tgctcaacat ttcctgggga tatctgttcg gaagcactgt atggctgtgc 180
ttcttcagtg agattgggtt ggtcaggaga atcaatgctc ctaaacggaa gaatctgcca 240
gagaatgcag aacaagccaa agaacagctg aaggagatca agaacaacga aggcgatttt 300
aaccgacgca atatcgactt caagtacttc tttagccttt ccacaatctt ctctagcata 360
ctgctgctta gcacagtgaa actcgccaac aacaatctgc agttgaggat ctgttccacc 420
attgtgtcac tgagttgcat actgaacaat atgtactttc agaacaagat acactcactt 480
gcactgaaga aagagagtct ctttaaggac atgatcgatc gtccgaaaga taccactatt 540
ctggtgaacc tgaagaagaa caagaccgac tttcacatcc atcatggcct ttctctgctc 600
ttgctctata gcagcttctt tggcctcact ccctacattt tcacgtgact cgag 654
<210> 67
<211> 211
<212> PRT
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> Ag40
<400> 67
Met Ala Gly Ala Ser Met Ser His Leu Gln Cys Leu Thr Ser Val Ala
1 5 10 15
Gly Leu Ser Ser Ile Val Met Ser Met Phe Pro Lys Leu Ile Ala Asn
20 25 30
Asn Pro Ser Leu Phe Arg Pro Leu Leu Asn Ile Ser Trp Gly Tyr Leu
35 40 45
Phe Gly Ser Thr Val Trp Leu Cys Phe Phe Ser Glu Ile Gly Leu Val
50 55 60
Arg Arg Ile Asn Ala Pro Lys Arg Lys Asn Leu Pro Glu Asn Ala Glu
65 70 75 80
Gln Ala Lys Glu Gln Leu Lys Glu Ile Lys Asn Asn Glu Gly Asp Phe
85 90 95
Asn Arg Arg Asn Ile Asp Phe Lys Tyr Phe Phe Ser Leu Ser Thr Ile
100 105 110
Phe Ser Ser Ile Leu Leu Leu Ser Thr Val Lys Leu Ala Asn Asn Asn
115 120 125
Leu Gln Leu Arg Ile Cys Ser Thr Ile Val Ser Leu Ser Cys Ile Leu
130 135 140
Asn Asn Met Tyr Phe Gln Asn Lys Ile His Ser Leu Ala Leu Lys Lys
145 150 155 160
Glu Ser Leu Phe Lys Asp Met Ile Asp Arg Pro Lys Asp Thr Thr Ile
165 170 175
Leu Val Asn Leu Lys Lys Asn Lys Thr Asp Phe His Ile His His Gly
180 185 190
Leu Ser Leu Leu Leu Leu Tyr Ser Ser Phe Phe Gly Leu Thr Pro Tyr
195 200 205
Ile Phe Thr
210
<210> 68
<211> 210
<212> PRT
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> Ag40
<400> 68
Met Gly Ala Ser Met Ser His Leu Gln Cys Leu Thr Ser Val Ala Gly
1 5 10 15
Leu Ser Ser Ile Val Met Ser Met Phe Pro Lys Leu Ile Ala Asn Asn
20 25 30
Pro Ser Leu Phe Arg Pro Leu Leu Asn Ile Ser Trp Gly Tyr Leu Phe
35 40 45
Gly Ser Thr Val Trp Leu Cys Phe Phe Ser Glu Ile Gly Leu Val Arg
50 55 60
Arg Ile Asn Ala Pro Lys Arg Lys Asn Leu Pro Glu Asn Ala Glu Gln
65 70 75 80
Ala Lys Glu Gln Leu Lys Glu Ile Lys Asn Asn Glu Gly Asp Phe Asn
85 90 95
Arg Arg Asn Ile Asp Phe Lys Tyr Phe Phe Ser Leu Ser Thr Ile Phe
100 105 110
Ser Ser Ile Leu Leu Leu Ser Thr Val Lys Leu Ala Asn Asn Asn Leu
115 120 125
Gln Leu Arg Ile Cys Ser Thr Ile Val Ser Leu Ser Cys Ile Leu Asn
130 135 140
Asn Met Tyr Phe Gln Asn Lys Ile His Ser Leu Ala Leu Lys Lys Glu
145 150 155 160
Ser Leu Phe Lys Asp Met Ile Asp Arg Pro Lys Asp Thr Thr Ile Leu
165 170 175
Val Asn Leu Lys Lys Asn Lys Thr Asp Phe His Ile His His Gly Leu
180 185 190
Ser Leu Leu Leu Leu Tyr Ser Ser Phe Phe Gly Leu Thr Pro Tyr Ile
195 200 205
Phe Thr
210
<210> 69
<211> 642
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> PfAg40 人 CO
<400> 69
gccaccatgg ctgggtgtac agtctctaat ctcaaatgcg tgaccaatgt ggcaggactg 60
gcaagtctgg ttatcagtct gtttccgaaa ctcatcataa agaacccaca agtgcttcga 120
ccactgctga atgtgtcctg gggttatctg tttggtagca ccttttggct gtgcttcttc 180
tccgaagtag gactgcttcg cagcctgaag aacatgaaag gggtaccttt gcctgaatca 240
gccagtgagg cgaagaagct tctcgaagag atgaagaact ctgagggcga tttcaatcgg 300
agatcactgg acttccagta cttcttttcc ctcgctacgt tgttctcagg cattctgttg 360
ctgagcacag tgaagttggc caaccataac ctgcagctta ggcttagtag ctctgtggtc 420
gtcatcacat cactgctgaa tagcctgtat ctgcacaata aagtgcataa tctgaaaagc 480
aagaaagaaa gcctctataa cgactttatt gccaatccca agaacgagaa aactgtcgct 540
gatctgaaga agaacaagaa agagtttcac atctttcacg gattgtccgt tctctctctc 600
tacgtttcct tcttcggcct gactccctac attttcacct aa 642
<210> 70
<211> 211
<212> PRT
<213> 恶性疟原虫（Plasmodium falciparum）
<220>
<221> misc_feature
<223> PfAg40
<400> 70
Met Ala Gly Cys Thr Val Ser Asn Leu Lys Cys Val Thr Asn Val Ala
1 5 10 15
Gly Leu Ala Ser Leu Val Ile Ser Leu Phe Pro Lys Leu Ile Ile Lys
20 25 30
Asn Pro Gln Val Leu Arg Pro Leu Leu Asn Val Ser Trp Gly Tyr Leu
35 40 45
Phe Gly Ser Thr Phe Trp Leu Cys Phe Phe Ser Glu Val Gly Leu Leu
50 55 60
Arg Ser Leu Lys Asn Met Lys Gly Val Pro Leu Pro Glu Ser Ala Ser
65 70 75 80
Glu Ala Lys Lys Leu Leu Glu Glu Met Lys Asn Ser Glu Gly Asp Phe
85 90 95
Asn Arg Arg Ser Leu Asp Phe Gln Tyr Phe Phe Ser Leu Ala Thr Leu
100 105 110
Phe Ser Gly Ile Leu Leu Leu Ser Thr Val Lys Leu Ala Asn His Asn
115 120 125
Leu Gln Leu Arg Leu Ser Ser Ser Val Val Val Ile Thr Ser Leu Leu
130 135 140
Asn Ser Leu Tyr Leu His Asn Lys Val His Asn Leu Lys Ser Lys Lys
145 150 155 160
Glu Ser Leu Tyr Asn Asp Phe Ile Ala Asn Pro Lys Asn Glu Lys Thr
165 170 175
Val Ala Asp Leu Lys Lys Asn Lys Lys Glu Phe His Ile Phe His Gly
180 185 190
Leu Ser Val Leu Ser Leu Tyr Val Ser Phe Phe Gly Leu Thr Pro Tyr
195 200 205
Ile Phe Thr
210
<210> 71
<211> 210
<212> PRT
<213> 恶性疟原虫（Plasmodium falciparum）
<220>
<221> misc_feature
<223> Ag40
<400> 71
Met Gly Cys Thr Val Ser Asn Leu Lys Cys Val Thr Asn Val Ala Gly
1 5 10 15
Leu Ala Ser Leu Val Ile Ser Leu Phe Pro Lys Leu Ile Ile Lys Asn
20 25 30
Pro Gln Val Leu Arg Pro Leu Leu Asn Val Ser Trp Gly Tyr Leu Phe
35 40 45
Gly Ser Thr Phe Trp Leu Cys Phe Phe Ser Glu Val Gly Leu Leu Arg
50 55 60
Ser Leu Lys Asn Met Lys Gly Val Pro Leu Pro Glu Ser Ala Ser Glu
65 70 75 80
Ala Lys Lys Leu Leu Glu Glu Met Lys Asn Ser Glu Gly Asp Phe Asn
85 90 95
Arg Arg Ser Leu Asp Phe Gln Tyr Phe Phe Ser Leu Ala Thr Leu Phe
100 105 110
Ser Gly Ile Leu Leu Leu Ser Thr Val Lys Leu Ala Asn His Asn Leu
115 120 125
Gln Leu Arg Leu Ser Ser Ser Val Val Val Ile Thr Ser Leu Leu Asn
130 135 140
Ser Leu Tyr Leu His Asn Lys Val His Asn Leu Lys Ser Lys Lys Glu
145 150 155 160
Ser Leu Tyr Asn Asp Phe Ile Ala Asn Pro Lys Asn Glu Lys Thr Val
165 170 175
Ala Asp Leu Lys Lys Asn Lys Lys Glu Phe His Ile Phe His Gly Leu
180 185 190
Ser Val Leu Ser Leu Tyr Val Ser Phe Phe Gly Leu Thr Pro Tyr Ile
195 200 205
Phe Thr
210
<210> 72
<211> 639
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> PvAg40 人 CO + kozac
<400> 72
gccaccatgg gggcgacggt atcatatctt agatgcgtga ccagtatagc agggctgagc 60
agcctcgtgc tgtctttgtt tcccaagctg attatgaaaa accctcaggt actccgacca 120
ctccttaaca ttagctgggg ttatttgttt ggctcaacct tttggttgtg tttgttctcc 180
gaagtaggac ttttccggtc cctgaaaaat atgaagcgca taccaatccc tgaaaacgca 240
gaagaggcta agaagcaatt ggaggagatg aaaagcatgg aaggggattt taccaggcgc 300
agggaagatt tccaatattt ttttggtttt tccaccttgt tttctggtat tcttcttctc 360
agtacggtaa gacttgcgaa tcacaacatg caactgagga tttccagtac catcgttgcc 420
cttagctgcc tgctcaataa cttgtacctt cagaataagg tacattctct taaaatccaa 480
aaagaaaacc tgtacaacga actcatccgc aatcctaagt cagagacgac tatagcggag 540
attaagaaaa acaaaaaaga tttccatata taccacggct tgtccctgtt gtccctttac 600
ataagcttcc tcggccttac tccatatata tttacctag 639
<210> 73
<211> 210
<212> PRT
<213> 间日疟原虫（Plasmodium vivax）
<220>
<221> misc_feature
<223> PvAg40
<400> 73
Met Gly Ala Thr Val Ser Tyr Leu Arg Cys Val Thr Ser Ile Ala Gly
1 5 10 15
Leu Ser Ser Leu Val Leu Ser Leu Phe Pro Lys Leu Ile Met Lys Asn
20 25 30
Pro Gln Val Leu Arg Pro Leu Leu Asn Ile Ser Trp Gly Tyr Leu Phe
35 40 45
Gly Ser Thr Phe Trp Leu Cys Leu Phe Ser Glu Val Gly Leu Phe Arg
50 55 60
Ser Leu Lys Asn Met Lys Arg Ile Pro Ile Pro Glu Asn Ala Glu Glu
65 70 75 80
Ala Lys Lys Gln Leu Glu Glu Met Lys Ser Met Glu Gly Asp Phe Thr
85 90 95
Arg Arg Arg Glu Asp Phe Gln Tyr Phe Phe Gly Phe Ser Thr Leu Phe
100 105 110
Ser Gly Ile Leu Leu Leu Ser Thr Val Arg Leu Ala Asn His Asn Met
115 120 125
Gln Leu Arg Ile Ser Ser Thr Ile Val Ala Leu Ser Cys Leu Leu Asn
130 135 140
Asn Leu Tyr Leu Gln Asn Lys Val His Ser Leu Lys Ile Gln Lys Glu
145 150 155 160
Asn Leu Tyr Asn Glu Leu Ile Arg Asn Pro Lys Ser Glu Thr Thr Ile
165 170 175
Ala Glu Ile Lys Lys Asn Lys Lys Asp Phe His Ile Tyr His Gly Leu
180 185 190
Ser Leu Leu Ser Leu Tyr Ile Ser Phe Leu Gly Leu Thr Pro Tyr Ile
195 200 205
Phe Thr
210
<210> 74
<211> 210
<212> PRT
<213> 间日疟原虫（Plasmodium vivax）
<220>
<221> misc_feature
<223> Ag40
<400> 74
Met Gly Ala Thr Val Ser Tyr Leu Arg Cys Val Thr Ser Ile Ala Gly
1 5 10 15
Leu Ser Ser Leu Val Leu Ser Leu Phe Pro Lys Leu Ile Met Lys Asn
20 25 30
Pro Gln Val Leu Arg Pro Leu Leu Asn Ile Ser Trp Gly Tyr Leu Phe
35 40 45
Gly Ser Thr Phe Trp Leu Cys Leu Phe Ser Glu Val Gly Leu Phe Arg
50 55 60
Ser Leu Lys Asn Met Lys Arg Ile Pro Ile Pro Glu Asn Ala Glu Glu
65 70 75 80
Ala Lys Lys Gln Leu Glu Glu Met Lys Ser Met Glu Gly Asp Phe Thr
85 90 95
Arg Arg Arg Glu Asp Phe Gln Tyr Phe Phe Gly Phe Ser Thr Leu Phe
100 105 110
Ser Gly Ile Leu Leu Leu Ser Thr Val Arg Leu Ala Asn His Asn Met
115 120 125
Gln Leu Arg Ile Ser Ser Thr Ile Val Ala Leu Ser Cys Leu Leu Asn
130 135 140
Asn Leu Tyr Leu Gln Asn Lys Val His Ser Leu Lys Ile Gln Lys Glu
145 150 155 160
Asn Leu Tyr Asn Glu Leu Ile Arg Asn Pro Lys Ser Glu Thr Thr Ile
165 170 175
Ala Glu Ile Lys Lys Asn Lys Lys Asp Phe His Ile Tyr His Gly Leu
180 185 190
Ser Leu Leu Ser Leu Tyr Ile Ser Phe Leu Gly Leu Thr Pro Tyr Ile
195 200 205
Phe Thr
210
<210> 75
<211> 1077
<212> DNA
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> Ag45
<400> 75
ggatccgcca ccatggcttc ttactcaaac tcttccatta agcagaaatc cgatagtgtg 60
agtgtctaca atactcggac tggaaatgtc agtaaaactc gcttgatccg tctgcaaaat 120
gggcattacc gtagagtggt cgacattagc aataaggacg agaaggagat tctcttcagg 180
acatgtgctt gcgcttgtcc aacacctcga aatgaggaga cacgcaaaac ctatatgcca 240
cctctgaaca atgtgtctac cgtagcgtat agaaagcgga tctattcttc ctttgggaat 300
aaggacggta acgatacagg caacaacgag agcataacag aacatgagga cccgattagg 360
accttttccg aaacgacaag taggcaggaa agtaccatcg acgacaaaac ggagactagc 420
atcaatagca aggaaacaga tgatggcaac cagtttggaa ggttgtttga agaactggag 480
gagaaagagg atgaactgat tgaggaaaag gaggaggagc tgatagaaga gaaggaggag 540
gaacttatag aggagaaaga agaagagctg atcgaagaga aagaggaaat cacccctgag 600
aacaaaaccc tcataatgcc ctctaaaact ctgatgaagg gcattaagac caacatttac 660
ttcctgtcaa acaaggaaaa gatccaagtg cttatgtgct ataactacaa gtgtgatgcc 720
gttgtgttcg agaaagacac ctttctgcgc tatctctaca tcaagagcat caataatatc 780
atcctgaacg aaagaatgat tgaacagttg tgcaagaacg aaaacctgaa gtacatcctt 840
gcctgcaaca gcatagtggt tgaatcaagc gacttcatca aacccctgat cattgagttt 900
gagtcatcca cttccaagaa catcttcgta aagcacatta agcacaatag ccagaaagaa 960
atggacatca acaagttcaa cgagtatatg cgggatctca aaagcaatga gaagctcaga 1020
ctgaagaaag tcgagcgatt ccactctatt aatctggcag ccaagaaatg actcgag 1077
<210> 76
<211> 352
<212> PRT
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> Ag45
<400> 76
Met Ala Ser Tyr Ser Asn Ser Ser Ile Lys Gln Lys Ser Asp Ser Val
1 5 10 15
Ser Val Tyr Asn Thr Arg Thr Gly Asn Val Ser Lys Thr Arg Leu Ile
20 25 30
Arg Leu Gln Asn Gly His Tyr Arg Arg Val Val Asp Ile Ser Asn Lys
35 40 45
Asp Glu Lys Glu Ile Leu Phe Arg Thr Cys Ala Cys Ala Cys Pro Thr
50 55 60
Pro Arg Asn Glu Glu Thr Arg Lys Thr Tyr Met Pro Pro Leu Asn Asn
65 70 75 80
Val Ser Thr Val Ala Tyr Arg Lys Arg Ile Tyr Ser Ser Phe Gly Asn
85 90 95
Lys Asp Gly Asn Asp Thr Gly Asn Asn Glu Ser Ile Thr Glu His Glu
100 105 110
Asp Pro Ile Arg Thr Phe Ser Glu Thr Thr Ser Arg Gln Glu Ser Thr
115 120 125
Ile Asp Asp Lys Thr Glu Thr Ser Ile Asn Ser Lys Glu Thr Asp Asp
130 135 140
Gly Asn Gln Phe Gly Arg Leu Phe Glu Glu Leu Glu Glu Lys Glu Asp
145 150 155 160
Glu Leu Ile Glu Glu Lys Glu Glu Glu Leu Ile Glu Glu Lys Glu Glu
165 170 175
Glu Leu Ile Glu Glu Lys Glu Glu Glu Leu Ile Glu Glu Lys Glu Glu
180 185 190
Ile Thr Pro Glu Asn Lys Thr Leu Ile Met Pro Ser Lys Thr Leu Met
195 200 205
Lys Gly Ile Lys Thr Asn Ile Tyr Phe Leu Ser Asn Lys Glu Lys Ile
210 215 220
Gln Val Leu Met Cys Tyr Asn Tyr Lys Cys Asp Ala Val Val Phe Glu
225 230 235 240
Lys Asp Thr Phe Leu Arg Tyr Leu Tyr Ile Lys Ser Ile Asn Asn Ile
245 250 255
Ile Leu Asn Glu Arg Met Ile Glu Gln Leu Cys Lys Asn Glu Asn Leu
260 265 270
Lys Tyr Ile Leu Ala Cys Asn Ser Ile Val Val Glu Ser Ser Asp Phe
275 280 285
Ile Lys Pro Leu Ile Ile Glu Phe Glu Ser Ser Thr Ser Lys Asn Ile
290 295 300
Phe Val Lys His Ile Lys His Asn Ser Gln Lys Glu Met Asp Ile Asn
305 310 315 320
Lys Phe Asn Glu Tyr Met Arg Asp Leu Lys Ser Asn Glu Lys Leu Arg
325 330 335
Leu Lys Lys Val Glu Arg Phe His Ser Ile Asn Leu Ala Ala Lys Lys
340 345 350
<210> 77
<211> 351
<212> PRT
<213> 伯氏疟原虫（Plasmodium berghei）
<220>
<221> misc_feature
<223> Ag45
<400> 77
Met Ser Tyr Ser Asn Ser Ser Ile Lys Gln Lys Ser Asp Ser Val Ser
1 5 10 15
Val Tyr Asn Thr Arg Thr Gly Asn Val Ser Lys Thr Arg Leu Ile Arg
20 25 30
Leu Gln Asn Gly His Tyr Arg Arg Val Val Asp Ile Ser Asn Lys Asp
35 40 45
Glu Lys Glu Ile Leu Phe Arg Thr Cys Ala Cys Ala Cys Pro Thr Pro
50 55 60
Arg Asn Glu Glu Thr Arg Lys Thr Tyr Met Pro Pro Leu Asn Asn Val
65 70 75 80
Ser Thr Val Ala Tyr Arg Lys Arg Ile Tyr Ser Ser Phe Gly Asn Lys
85 90 95
Asp Gly Asn Asp Thr Gly Asn Asn Glu Ser Ile Thr Glu His Glu Asp
100 105 110
Pro Ile Arg Thr Phe Ser Glu Thr Thr Ser Arg Gln Glu Ser Thr Ile
115 120 125
Asp Asp Lys Thr Glu Thr Ser Ile Asn Ser Lys Glu Thr Asp Asp Gly
130 135 140
Asn Gln Phe Gly Arg Leu Phe Glu Glu Leu Glu Glu Lys Glu Asp Glu
145 150 155 160
Leu Ile Glu Glu Lys Glu Glu Glu Leu Ile Glu Glu Lys Glu Glu Glu
165 170 175
Leu Ile Glu Glu Lys Glu Glu Glu Leu Ile Glu Glu Lys Glu Glu Ile
180 185 190
Thr Pro Glu Asn Lys Thr Leu Ile Met Pro Ser Lys Thr Leu Met Lys
195 200 205
Gly Ile Lys Thr Asn Ile Tyr Phe Leu Ser Asn Lys Glu Lys Ile Gln
210 215 220
Val Leu Met Cys Tyr Asn Tyr Lys Cys Asp Ala Val Val Phe Glu Lys
225 230 235 240
Asp Thr Phe Leu Arg Tyr Leu Tyr Ile Lys Ser Ile Asn Asn Ile Ile
245 250 255
Leu Asn Glu Arg Met Ile Glu Gln Leu Cys Lys Asn Glu Asn Leu Lys
260 265 270
Tyr Ile Leu Ala Cys Asn Ser Ile Val Val Glu Ser Ser Asp Phe Ile
275 280 285
Lys Pro Leu Ile Ile Glu Phe Glu Ser Ser Thr Ser Lys Asn Ile Phe
290 295 300
Val Lys His Ile Lys His Asn Ser Gln Lys Glu Met Asp Ile Asn Lys
305 310 315 320
Phe Asn Glu Tyr Met Arg Asp Leu Lys Ser Asn Glu Lys Leu Arg Leu
325 330 335
Lys Lys Val Glu Arg Phe His Ser Ile Asn Leu Ala Ala Lys Lys
340 345 350
<210> 78
<211> 1197
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> PfAg45 人 CO
<400> 78
gccaccatgg cttcagatta ctttacaatt ctgtccaata ttttcacaag cactagcctg 60
aagaagaaat acagttcccg gttgagcaca aaatccaaga agaaccaaaa gcgagtcaaa 120
ctgataagac tgcggaatgg acattttcgc cgaattgtgg atatttccaa cattgacgag 180
aagagcatct tccccagaag ctgtactttt gcgtcaatta gcagtgctag caaagaaaac 240
gagaggaaga attcaagcga ggacacaaaa gaacctcagg agaatctgta tggcaaatca 300
aacacttcaa gctctatcac gataaagatc aatttcgacg aaagcgatga gaacaagagt 360
gatcaggata accactctat cgataccatt agcgacatct cttttaccca gacttcacgc 420
aaatctcttg aaattgaaag taatacctat gagagttatc gcgaagtgga gaaggaggac 480
attgaggagg aggaggaaga ggagaaagaa gaagaatatg aggaggaaga agaagaagag 540
gaatacgaag aggaagagga agaagaggag gaagagtatg aggaggaggg tctgaaaacc 600
gaagaggaga aggaagaaga taataaggag gtagagccag aagaggagct taaagaagaa 660
gatgacaagg aggttgagcc tgaggaggag aaggagaatg agcagaagaa agaagaacaa 720
gaggagaata acctcgaagc tcccagcaaa acactgatga aaggggttaa gaccaacata 780
tacttcctgt ctaccaaaga gcggatagaa gcactcatgt gctacaacta catatccaac 840
gccattattt tcgaaaaggg caagtttctc cgttatatct tcatgaacaa tgtcaacaat 900
atcatcgtga acgagcacat gatcaatatg ttgtgcaaga aggaaaagat caaatacatc 960
ctgtcatcta actccatcat cattgaaagc aacgacttca tcaaaccgct catcattgag 1020
tttgacagta acatctctaa gaagatcttt gtcaaacact tgaaaatggt ggactccttc 1080
aaactggatg acaagctgta cagggagtac ctgaatgacc tttctgaaca tgagagggat 1140
agactgaaac atgtggagtc cttctattcc aatgccataa aggtgcacaa tacgtaa 1197
<210> 79
<211> 396
<212> PRT
<213> 恶性疟原虫（Plasmodium falciparum）
<220>
<221> misc_feature
<223> PfAg45
<400> 79
Met Ala Ser Asp Tyr Phe Thr Ile Leu Ser Asn Ile Phe Thr Ser Thr
1 5 10 15
Ser Leu Lys Lys Lys Tyr Ser Ser Arg Leu Ser Thr Lys Ser Lys Lys
20 25 30
Asn Gln Lys Arg Val Lys Leu Ile Arg Leu Arg Asn Gly His Phe Arg
35 40 45
Arg Ile Val Asp Ile Ser Asn Ile Asp Glu Lys Ser Ile Phe Pro Arg
50 55 60
Ser Cys Thr Phe Ala Ser Ile Ser Ser Ala Ser Lys Glu Asn Glu Arg
65 70 75 80
Lys Asn Ser Ser Glu Asp Thr Lys Glu Pro Gln Glu Asn Leu Tyr Gly
85 90 95
Lys Ser Asn Thr Ser Ser Ser Ile Thr Ile Lys Ile Asn Phe Asp Glu
100 105 110
Ser Asp Glu Asn Lys Ser Asp Gln Asp Asn His Ser Ile Asp Thr Ile
115 120 125
Ser Asp Ile Ser Phe Thr Gln Thr Ser Arg Lys Ser Leu Glu Ile Glu
130 135 140
Ser Asn Thr Tyr Glu Ser Tyr Arg Glu Val Glu Lys Glu Asp Ile Glu
145 150 155 160
Glu Glu Glu Glu Glu Glu Lys Glu Glu Glu Tyr Glu Glu Glu Glu Glu
165 170 175
Glu Glu Glu Tyr Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Tyr Glu
180 185 190
Glu Glu Gly Leu Lys Thr Glu Glu Glu Lys Glu Glu Asp Asn Lys Glu
195 200 205
Val Glu Pro Glu Glu Glu Leu Lys Glu Glu Asp Asp Lys Glu Val Glu
210 215 220
Pro Glu Glu Glu Lys Glu Asn Glu Gln Lys Lys Glu Glu Gln Glu Glu
225 230 235 240
Asn Asn Leu Glu Ala Pro Ser Lys Thr Leu Met Lys Gly Val Lys Thr
245 250 255
Asn Ile Tyr Phe Leu Ser Thr Lys Glu Arg Ile Glu Ala Leu Met Cys
260 265 270
Tyr Asn Tyr Ile Ser Asn Ala Ile Ile Phe Glu Lys Gly Lys Phe Leu
275 280 285
Arg Tyr Ile Phe Met Asn Asn Val Asn Asn Ile Ile Val Asn Glu His
290 295 300
Met Ile Asn Met Leu Cys Lys Lys Glu Lys Ile Lys Tyr Ile Leu Ser
305 310 315 320
Ser Asn Ser Ile Ile Ile Glu Ser Asn Asp Phe Ile Lys Pro Leu Ile
325 330 335
Ile Glu Phe Asp Ser Asn Ile Ser Lys Lys Ile Phe Val Lys His Leu
340 345 350
Lys Met Val Asp Ser Phe Lys Leu Asp Asp Lys Leu Tyr Arg Glu Tyr
355 360 365
Leu Asn Asp Leu Ser Glu His Glu Arg Asp Arg Leu Lys His Val Glu
370 375 380
Ser Phe Tyr Ser Asn Ala Ile Lys Val His Asn Thr
385 390 395
<210> 80
<211> 395
<212> PRT
<213> 恶性疟原虫（Plasmodium falciparum）
<220>
<221> misc_feature
<223> Ag45
<400> 80
Met Ser Asp Tyr Phe Thr Ile Leu Ser Asn Ile Phe Thr Ser Thr Ser
1 5 10 15
Leu Lys Lys Lys Tyr Ser Ser Arg Leu Ser Thr Lys Ser Lys Lys Asn
20 25 30
Gln Lys Arg Val Lys Leu Ile Arg Leu Arg Asn Gly His Phe Arg Arg
35 40 45
Ile Val Asp Ile Ser Asn Ile Asp Glu Lys Ser Ile Phe Pro Arg Ser
50 55 60
Cys Thr Phe Ala Ser Ile Ser Ser Ala Ser Lys Glu Asn Glu Arg Lys
65 70 75 80
Asn Ser Ser Glu Asp Thr Lys Glu Pro Gln Glu Asn Leu Tyr Gly Lys
85 90 95
Ser Asn Thr Ser Ser Ser Ile Thr Ile Lys Ile Asn Phe Asp Glu Ser
100 105 110
Asp Glu Asn Lys Ser Asp Gln Asp Asn His Ser Ile Asp Thr Ile Ser
115 120 125
Asp Ile Ser Phe Thr Gln Thr Ser Arg Lys Ser Leu Glu Ile Glu Ser
130 135 140
Asn Thr Tyr Glu Ser Tyr Arg Glu Val Glu Lys Glu Asp Ile Glu Glu
145 150 155 160
Glu Glu Glu Glu Glu Lys Glu Glu Glu Tyr Glu Glu Glu Glu Glu Glu
165 170 175
Glu Glu Tyr Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Tyr Glu Glu
180 185 190
Glu Gly Leu Lys Thr Glu Glu Glu Lys Glu Glu Asp Asn Lys Glu Val
195 200 205
Glu Pro Glu Glu Glu Leu Lys Glu Glu Asp Asp Lys Glu Val Glu Pro
210 215 220
Glu Glu Glu Lys Glu Asn Glu Gln Lys Lys Glu Glu Gln Glu Glu Asn
225 230 235 240
Asn Leu Glu Ala Pro Ser Lys Thr Leu Met Lys Gly Val Lys Thr Asn
245 250 255
Ile Tyr Phe Leu Ser Thr Lys Glu Arg Ile Glu Ala Leu Met Cys Tyr
260 265 270
Asn Tyr Ile Ser Asn Ala Ile Ile Phe Glu Lys Gly Lys Phe Leu Arg
275 280 285
Tyr Ile Phe Met Asn Asn Val Asn Asn Ile Ile Val Asn Glu His Met
290 295 300
Ile Asn Met Leu Cys Lys Lys Glu Lys Ile Lys Tyr Ile Leu Ser Ser
305 310 315 320
Asn Ser Ile Ile Ile Glu Ser Asn Asp Phe Ile Lys Pro Leu Ile Ile
325 330 335
Glu Phe Asp Ser Asn Ile Ser Lys Lys Ile Phe Val Lys His Leu Lys
340 345 350
Met Val Asp Ser Phe Lys Leu Asp Asp Lys Leu Tyr Arg Glu Tyr Leu
355 360 365
Asn Asp Leu Ser Glu His Glu Arg Asp Arg Leu Lys His Val Glu Ser
370 375 380
Phe Tyr Ser Asn Ala Ile Lys Val His Asn Thr
385 390 395
<210> 81
<211> 1122
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> PvAg45 人 CO + kozac
<400> 81
gccaccatgg ctaatttgag ttctcccctc ctggccttgc ccgaggaggg taagaagcga 60
agaacaaaac tcatccgact gaggaacggt cactataggc ggatagtgga catttcaaat 120
accgacgaac ggaagctcat tccttctatg tgccgctgcg cgtgtgtcac tcccagaaaa 180
gacgaagtgg agaatgaggg taagtgggaa gacgctaaga aagcaaaatc aagccaggaa 240
tatgatgaaa cttctgatta tgttgaatct gaaaagaaag agagctatat gctcgcagtc 300
aatgaggagg atcagacgga ggatatgtac tcaaagacga tcagctttac ttctataacc 360
cctacatcta taagatccga agagccagag ccaaggcgga aactctccct tctggatgtt 420
aaagaagaag aggaagagga ggaggaggaa gaagaagagg aagaagagga agaggaggaa 480
gaggaagagg aagaagagga agagaaagaa aaagaaaaag agaaggagga agaggaagaa 540
gaggaggagg aagaggaaga ggaagaggag gaggaagaag aagaggaaga tgaaatagaa 600
tctaccgcag aggaaaagga agaagagaag aagcaagtcc caccggaagg taagaaattg 660
atcgaaccct caaagaccct tatgagaggg actaagacca acatttattt tttgagtaat 720
aaggagatgg ttcaaactct gatgtgttat aattataatt gcaacgcagt ggtattcgaa 780
aaagacactt ttttgaggta tctgtacatg aagagcatca gcaacatcat cctcaacgaa 840
cgaatgatag atgaactgtg caaacaggaa gatcttaaat acgtgcttac aagcaatgct 900
atcgtgttgg aatctactga ctttcttaag cctcttataa ttgagtttga atccagtatt 960
agtaaaagag ttttcgtgcg ccacctgaag cataacgctc gcaaggaaat cgacatgaaa 1020
aaatatcatg attacatggg cgaacttaac gctaacgaga agattaggct gatgaaaatt 1080
gagcgatttc atagtttcaa caagatgatc caatgcaact ag 1122
<210> 82
<211> 371
<212> PRT
<213> 间日疟原虫（Plasmodium vivax）
<220>
<221> misc_feature
<223> PvAg45
<400> 82
Met Ala Asn Leu Ser Ser Pro Leu Leu Ala Leu Pro Glu Glu Gly Lys
1 5 10 15
Lys Arg Arg Thr Lys Leu Ile Arg Leu Arg Asn Gly His Tyr Arg Arg
20 25 30
Ile Val Asp Ile Ser Asn Thr Asp Glu Arg Lys Leu Ile Pro Ser Met
35 40 45
Cys Arg Cys Ala Cys Val Thr Pro Arg Lys Asp Glu Val Glu Asn Glu
50 55 60
Gly Lys Trp Glu Asp Ala Lys Lys Ala Lys Ser Ser Gln Glu Tyr Asp
65 70 75 80
Glu Thr Ser Asp Tyr Val Glu Ser Glu Lys Lys Glu Ser Tyr Met Leu
85 90 95
Ala Val Asn Glu Glu Asp Gln Thr Glu Asp Met Tyr Ser Lys Thr Ile
100 105 110
Ser Phe Thr Ser Ile Thr Pro Thr Ser Ile Arg Ser Glu Glu Pro Glu
115 120 125
Pro Arg Arg Lys Leu Ser Leu Leu Asp Val Lys Glu Glu Glu Glu Glu
130 135 140
Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu
145 150 155 160
Glu Glu Glu Glu Glu Glu Lys Glu Lys Glu Lys Glu Lys Glu Glu Glu
165 170 175
Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu
180 185 190
Glu Glu Asp Glu Ile Glu Ser Thr Ala Glu Glu Lys Glu Glu Glu Lys
195 200 205
Lys Gln Val Pro Pro Glu Gly Lys Lys Leu Ile Glu Pro Ser Lys Thr
210 215 220
Leu Met Arg Gly Thr Lys Thr Asn Ile Tyr Phe Leu Ser Asn Lys Glu
225 230 235 240
Met Val Gln Thr Leu Met Cys Tyr Asn Tyr Asn Cys Asn Ala Val Val
245 250 255
Phe Glu Lys Asp Thr Phe Leu Arg Tyr Leu Tyr Met Lys Ser Ile Ser
260 265 270
Asn Ile Ile Leu Asn Glu Arg Met Ile Asp Glu Leu Cys Lys Gln Glu
275 280 285
Asp Leu Lys Tyr Val Leu Thr Ser Asn Ala Ile Val Leu Glu Ser Thr
290 295 300
Asp Phe Leu Lys Pro Leu Ile Ile Glu Phe Glu Ser Ser Ile Ser Lys
305 310 315 320
Arg Val Phe Val Arg His Leu Lys His Asn Ala Arg Lys Glu Ile Asp
325 330 335
Met Lys Lys Tyr His Asp Tyr Met Gly Glu Leu Asn Ala Asn Glu Lys
340 345 350
Ile Arg Leu Met Lys Ile Glu Arg Phe His Ser Phe Asn Lys Met Ile
355 360 365
Gln Cys Asn
370
<210> 83
<211> 345
<212> PRT
<213> 间日疟原虫（Plasmodium vivax）
<220>
<221> misc_feature
<223> Ag45
<400> 83
Met Asn Leu Ser Ser Pro Leu Leu Ala Leu Pro Glu Glu Gly Lys Lys
1 5 10 15
Arg Arg Thr Lys Leu Ile Arg Leu Arg Asn Gly His Tyr Arg Arg Ile
20 25 30
Val Asp Ile Ser Asn Thr Asp Glu Arg Lys Leu Ile Pro Ser Met Cys
35 40 45
Arg Cys Ala Cys Val Thr Pro Arg Lys Asp Glu Val Glu Asn Glu Gly
50 55 60
Lys Trp Glu Asp Ala Lys Lys Ala Lys Ser Ser Gln Glu Tyr Asp Glu
65 70 75 80
Thr Ser Asp Tyr Val Glu Ser Glu Lys Lys Glu Ser Tyr Met Leu Ala
85 90 95
Val Asn Glu Glu Asp Gln Thr Glu Asp Met Tyr Ser Lys Thr Ile Ser
100 105 110
Phe Thr Ser Ile Thr Pro Thr Ser Ile Arg Ser Glu Glu Pro Glu Pro
115 120 125
Arg Arg Lys Leu Ser Leu Leu Asp Val Lys Glu Glu Glu Glu Glu Glu
130 135 140
Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu
145 150 155 160
Glu Lys Glu Glu Glu Glu Glu Glu Asp Glu Ile Glu Ser Thr Ala Glu
165 170 175
Glu Lys Glu Glu Glu Lys Lys Gln Val Pro Pro Glu Gly Lys Lys Leu
180 185 190
Ile Glu Pro Ser Lys Thr Leu Met Arg Gly Thr Lys Thr Asn Ile Tyr
195 200 205
Phe Leu Ser Asn Lys Glu Met Val Gln Thr Leu Met Cys Tyr Asn Tyr
210 215 220
Asn Cys Asn Ala Val Val Phe Glu Lys Asp Thr Phe Leu Arg Tyr Leu
225 230 235 240
Tyr Met Lys Ser Ile Ser Asn Ile Ile Leu Asn Glu Arg Met Ile Asp
245 250 255
Glu Leu Cys Lys Gln Glu Asp Leu Lys Tyr Val Leu Thr Ser Asn Ala
260 265 270
Ile Val Leu Glu Ser Thr Asp Phe Leu Lys Pro Leu Ile Ile Glu Phe
275 280 285
Glu Ser Ser Ile Ser Lys Arg Val Phe Val Arg His Leu Lys His Asn
290 295 300
Ala Arg Lys Glu Ile Asp Met Lys Lys Tyr His Asp Tyr Met Gly Glu
305 310 315 320
Leu Asn Ala Asn Glu Lys Ile Arg Leu Met Lys Ile Glu Arg Phe His
325 330 335
Ser Phe Asn Lys Met Ile Gln Cys Asn
340 345
<210> 84
<211> 12
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> Kozac共有序列
<400> 84
gccaccatgg ct 12
<210> 85
<211> 12
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> Kozac共有序列
<400> 85
gccaccatgg cc 12
<210> 86
<211> 6
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> BamHI位点
<400> 86
ggatcc 6
<210> 87
<211> 6
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> Xhol位点
<400> 87
ctcgag 6
<210> 88
<211> 9
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> CD8 T细胞表位
<400> 88
Gln Ala Gln Arg Asn Leu Asn Glu Leu
1 5
<210> 89
<211> 9
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> CD8 T细胞表位
<400> 89
Ser Ala Leu Leu Asn Val Asp Asn Leu
1 5
<210> 90
<211> 9
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> CD8 T细胞表位
<400> 90
Lys Ser Pro Ser Asn Phe Thr Ile Ile
1 5
<210> 91
<211> 9
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> CD8 T细胞表位
<400> 91
Ser Asn Gln Thr Asn Gln Glu Thr Ile
1 5
<210> 92
<211> 9
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> CD8 T细胞表位
<400> 92
Ile Thr Pro Glu Asn Lys Thr Leu Ile
1 5
<210> 93
<211> 9
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> CD8 T细胞表位
<400> 93
Lys Leu Ile Ala Asn Asn Pro Ser Leu
1 5
<210> 94
<211> 9
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> CD8 T细胞表位
<400> 94
Ser Cys Ile Leu Asn Asn Met Tyr Phe
1 5
PCT/RO/134表

Claims (21)

1.化合物的组合物，其中，所述组合物包含至少两种不同的活性成分，其中，每种活性成分由疟原虫寄生虫的抗原多肽、编码所述抗原多肽的多核苷酸或载体组成，所述载体表达疟原虫寄生虫的抗原多肽，特别地，所述载体是病毒载体，尤其是慢病毒载体；其中，一种抗原多肽是环子孢子蛋白CSP或它的多肽衍生物，另一种抗原多肽是：蛋白Ag40(11-09)或它的多肽衍生物，或蛋白Ag45(11-10)或它的多肽衍生物，所述蛋白Ag40(11-09)具有SEQID NO：67、68、70、71、73或74的序列，所述蛋白Ag45(11-10)具有SEQ ID NO：76、77、79、80、82或83的序列，前提是每种多肽衍生物保持衍生其的抗原的保护性。
2.根据权利要求1所述的组合物，其中，所述组合物进一步包含以下物质作为活性成分：一种以上疟原虫寄生虫的抗原多肽、编码所述抗原多肽的多核苷酸，或表达所述疟原虫寄生虫的抗原多肽的载体，特别地，所述载体是病毒载体，尤其是慢病毒载体；其中，每种抗原多肽选自：血小板反应蛋白相关匿名蛋白TRAP、半胱氨酸蛋白酶抑制剂ICP、金属肽酶Falcilysin、GPI锚定蛋白P113、成孔样蛋白SPECT2或任何这些抗原多肽的多肽衍生物，其中，所述多肽衍生物基本上保持衍生其的抗原的保护性。
3.根据权利要求1或2所述的化合物的组合物，其中，所述组合物包含一组不同的活性成分，其中，每种活性成分由疟原虫寄生虫的抗原多肽、编码所述抗原多肽的多核苷酸或表达所述疟原虫寄生虫的抗原多肽的载体组成，特别地，所述载体是病毒载体，尤其是慢病毒载体；其中，所述抗原多肽的组包含疟原虫寄生虫的红前期抗原，所述抗原包括：环子孢子蛋白CSP、血小板反应蛋白相关匿名蛋白TRAP、半胱氨酸蛋白酶抑制剂ICP、金属肽酶Falcilysin、GPI锚定蛋白P113、成孔样蛋白SPECT2、具有SEQ ID NO：67、68、70、71、73或74序列的蛋白Ag40(11-09)或它的变体、具有SEQ ID NO：76、77、79、80、82或83序列的蛋白Ag45(11-10)或它的变体，或任何这些抗原多肽的多肽衍生物，其中，所述多肽衍生物保持衍生其的抗原的保护性。
4.根据权利要求1至3中任一项所述的化合物的组合物，其中，在所述化合物的组合物中，疟原虫的不同活性成分以单独的活性成分提供，或以多种活性成分的单独组合物提供。
5.根据权利要求1至4中任一项所述的化合物的组合物，其中，疟原虫的不同活性成分以单一组合物提供。
6.根据权利要求1至5中任一项所述的化合物的组合物，其中，所述活性成分由人感染疟原虫寄生虫的抗原多肽组成，或所述活性成分由表达人感染疟原虫寄生虫的抗原多肽的慢病毒载体组成，或所述活性成分包含这些抗原多肽和病毒载体的混合物或结合物，特别地，其中疟原虫寄生虫是恶性疟原虫或间日疟原虫。
7.根据权利要求1至6中任一项所述的化合物的组合物，其中，所述活性成分包含以下抗原或由以下抗原组成：序列为SEQ ID NO：11、12、14、15、17或18的环子孢子蛋白CSP；序列为SEQ ID NO：20、21、23、24、26或27的血小板反应蛋白相关匿名蛋白TRAP；序列为SEQ IDNO：29、30、32、33、35或36的半胱氨酸蛋白酶抑制剂ICP；序列为SEQ ID NO：38、39、41、42、44、45或47的金属肽酶Falcilysin；序列为SEQ ID NO：58、59、61、62、64或65的GPI锚定蛋白P113；序列为SEQ ID NO：49、50、52、53、55或56的成孔样蛋白SPECT2；序列为SEQ ID NO：67、68、70、71、73或74的蛋白Ag40(11-09)或它的变体；以及序列为SEQ ID NO.76、77、79、80、82或83的蛋白Ag45(11-10)或它的变体；或它们的多肽衍生物，所述多肽衍生物由与衍生其的抗原多肽具有至少70％氨基酸同一性的序列组成，优选由与衍生其的抗原多肽具有至少95％的氨基酸同一性的序列组成，并且所述多肽衍生物保持衍生其的多肽的保护性。
8.根据权利要求1至7中任一项所述的化合物的组合物，其中，所述活性成分包含人慢病毒载体或由人慢病毒载体组成，所述人慢病毒载体表达所述抗原多肽或它的多肽衍生物，特别地，所述载体是HIV-1慢病毒载体，其中，所述抗原多肽或它的多肽衍生物：
-由单独的载体单个地表达，和/或
-由一种以上载体表达，其中，至少一种载体表达超过一种的抗原多肽或它的多肽衍生物。
9.根据权利要求8所述的化合物的组合物，其中，每种慢病毒载体是复制缺陷的假型慢病毒载体，特别是复制缺陷的假型HIV-1慢病毒载体，其中，所述载体包含基因组，所述基因组包含哺乳动物密码子优化的合成核酸，特别是人类密码子优化的合成核酸，其中，所述合成核酸编码感染哺乳动物寄主的疟原虫寄生虫的抗原多肽或它的多肽衍生物，特别地，所述哺乳动物寄主是人类寄主。
10.根据权利要求8或9所述的化合物的组合物，其中，每种慢病毒载体是整合假型慢病毒载体和/或非整合假型慢病毒载体，特别是整合HIV-1载体，其中，所述载体包含基因组，所述基因组包含哺乳动物密码子优化的合成核酸，特别是人类密码子优化的合成核酸，其中，所述合成核酸编码感染哺乳动物寄主的疟原虫寄生虫的抗原多肽或它的多肽衍生物，特别地，所述哺乳动物寄主是人类寄主。
11.根据权利要求8至10所述的化合物的组合物，其中，慢病毒载体基因组的基因组由pTRIP载体质粒获得，其中，在哺乳动物细胞中的功能性启动子的控制下，特别是在人β-2微球蛋白启动子的控制下，可选地，在旱獭肝炎病毒WPRE的转录后调控元件的控制下，克隆编码抗原多肽或它的多肽衍生物的疟原虫合成核酸。
12.根据权利要求8至11中任一项所述的化合物的组合物，其中，慢病毒载体用来自印第安纳血清型或新泽西血清型水疱性口炎病毒V-SVG的糖蛋白G进行假型化。
13.根据权利要求1至12中任一项所述的化合物的组合物，其中，所述抗原多肽或它的多肽衍生物由核酸分子表达，所述核酸分子是哺乳动物密码子优化的合成核酸，所述核酸分子的特征在于具有以下序列：
-a.伯氏疟原虫的CSP的SEQ ID NO：10、伯氏疟原虫的TRAP的SEQ ID NO：19、伯氏疟原虫的ICP的SEQ ID NO：28、伯氏疟原虫的Falcilysin的SEQ ID NO：37和46、伯氏疟原虫的GPI锚定蛋白P113的SEQ ID NO：57、伯氏疟原虫的成孔样蛋白SPECT2的SEQ ID NO：48、伯氏疟原虫的蛋白Ag40 11-09的SEQ ID NO：66，以及伯氏疟原虫的蛋白Ag45 11-10的SEQ IDNO：75，或者，
-b.恶性疟原虫的CSP的SEQ ID NO：13、恶性疟原虫的TRAP的SEQ ID NO：22、恶性疟原虫的ICP的SEQ ID NO：31、恶性疟原虫的Falcilysin的SEQ ID NO：40、恶性疟原虫的成孔样蛋白SPECT2的SEQ ID NO：51、恶性疟原虫的GPI锚定蛋白P113的SEQ ID NO：60、恶性疟原虫的蛋白11-09的SEQ ID NO：69，以及恶性疟原虫的蛋白11-10的SEQ ID NO：78，或者，
-c.间日疟原虫的CSP的SEQ ID NO：16、间日疟原虫的TRAP的SEQ ID NO：25、间日疟原虫的ICP的SEQ ID NO：34、间日疟原虫的Falcilysin的SEQ ID NO：43、间日疟原虫的成孔样蛋白SPECT2的SEQ ID NO：54、间日疟原虫的GPI锚定蛋白P113的SEQ ID NO：63、间日疟原虫的蛋白11-09的SEQ ID NO：72，以及间日疟原虫的蛋白11-10的SEQ ID NO：81。
14.一种适用于给哺乳动物寄主施用的制剂，其中，所述制剂包含权利要求1至13中任一项所述的化合物的组合物作为活性成分，所述活性成分用于保护免受疟原虫感染或寄生虫诱导的病症或疾病，并且所述制剂包含适用于给有需要的寄主施用的赋形剂，所述寄主特别是人类寄主。
15.根据权利要求1至14中任一项所述的化合物的组合物，其中，所述化合物的组合物用于在哺乳动物寄主中，特别是在人类寄主中，针对疟疾寄生虫感染或针对寄生虫诱导的病症或疾病的保护性免疫，可选地与适当的递送载体和可选的佐剂组分和/或免疫刺激剂组分结合。
16.根据权利要求1至15中任一项所述的化合物的组合物，其中，所述化合物的组合物用于在哺乳动物寄主中，特别是在人类寄主中，针对疟疾寄生虫感染或针对寄生虫诱导的病症或疾病的保护性免疫，涉及免疫模式的用途包括：施用有效量的化合物，特别是慢病毒载体，引发寄主的细胞免疫应答，随后及时施用有效量的化合物，特别是慢病毒载体，加强寄主的细胞免疫应答，并可选地重复用于加强的施用步骤，其中，用不同的包膜蛋白假型化在每个引发步骤或加强步骤中施用的化合物，特别是慢病毒载体，所述包膜蛋白不进行交叉血清中和，并且，所述引发步骤和加强步骤在时间上分隔至少6周，特别是至少8周。
17.根据权利要求16所述的化合物的组合物，其中，所述化合物的组合物用于按照剂量方案在哺乳动物寄主中，特别是在人类寄主中，针对疟疾寄生虫感染或针对寄生虫诱导的病症或疾病的保护性免疫，所述剂量方案包括：单独提供的化合物剂量，所述化合物特别是慢病毒载体，其中，用于引发细胞免疫应答的剂量是中等剂量，用于加强细胞免疫应答的剂量高于引发剂量。
18.根据权利要求16或17中任一项所述的化合物的组合物，其中，所述化合物的组合物用于按照剂量方案在哺乳动物寄主中，特别是在人类寄主中，针对疟疾寄生虫感染或针对寄生虫诱导的病症或疾病的保护性免疫，所述剂量方案包括：单独提供的慢病毒载体剂量，其中，当使用整合型载体颗粒时，用于引发细胞免疫应答的剂量和用于加强细胞免疫应答的剂量各自包含105至109TU，特别是105至107TU；当使用整合缺陷型载体颗粒时，用于引发细胞免疫应答的剂量和用于加强细胞免疫应答的剂量包含107至1010TU。
19.权利要求1至18中任一项所述的化合物的组合物在制备用于给哺乳动物寄主注射的施用形式中的用途，其中，所述施用形式包含一剂量的所述化合物，所述化合物能够在哺乳动物寄主中，特别是在人类寄主中，产生针对疟疾寄生虫感染或针对寄生虫诱导的病症或疾病的保护性免疫。
20.一种多核苷酸，其中，所述多核苷酸是小鼠密码子优化的合成核酸，所述合成核酸编码疟原虫寄生虫的红前期抗原，其中，所述多核苷酸选自下组：
-a.伯氏疟原虫的CSP的SEQ ID NO：10、伯氏疟原虫的TRAP的SEQ ID NO：19、伯氏疟原虫的ICP的SEQ ID NO：28、伯氏疟原虫的Falcilysin的SEQ ID NO：37、伯氏疟原虫的GPI锚定蛋白P113的SEQ ID NO：57、伯氏疟原虫的成孔样蛋白SPECT2的SEQ ID NO：48、伯氏疟原虫的蛋白Ag40 11-09的SEQ ID NO：66，以及伯氏疟原虫的蛋白Ag45 11-10的SEQ ID NO：75，或者，
-b.恶性疟原虫的CSP的SEQ ID NO：13、恶性疟原虫的TRAP的SEQ ID NO：22、恶性疟原虫的ICP的SEQ ID NO：31、恶性疟原虫的Falcilysin的SEQ ID NO：40、恶性疟原虫的成孔样蛋白SPECT2的SEQ ID NO：51、恶性疟原虫的GPI锚定蛋白P113的SEQ ID NO：60、恶性疟原虫的蛋白11-09的SEQ ID NO：69，以及恶性疟原虫的蛋白11-10的SEQ ID NO：78，或者，
-c.间日疟原虫的CSP的SEQ ID NO：16、间日疟原虫的TRAP的SEQ ID NO：25、间日疟原虫的ICP的SEQ ID NO：34、间日疟原虫的Falcilysin的SEQ ID NO：43、间日疟原虫的成孔样蛋白SPECT2的SEQ ID NO：54、间日疟原虫的GPI锚定蛋白P113的SEQ ID NO：63、间日疟原虫的蛋白11-09的SEQ ID NO：72，以及间日疟原虫的蛋白11-10的SEQ ID NO：81。
21.权利要求20所述的多核苷酸在制备慢病毒载体的集合中的用途，特别地，所述慢病毒载体是基于HIV-1的载体，其中，载体在它的基因组中包含至少一种权利要求20所述的多核苷酸，前提是所述慢病毒载体的集合能够表达所有抗原多肽，所述抗原多肽由选自a/、b/和c/的一个亚组的多核苷酸编码。
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