一种抗CD19抗体及其制备方法和用途与流程

本申请为《一种抗cd19抗体及其制备方法和用途》的分案申请，原申请的申请日：2017.9.6，申请号：201710793590.1。本发明涉及生物
技术领域：
，特别是涉及一种抗cd19抗体及其制备方法和用途。
背景技术：
：cd19抗原(blymphocyteantigencd19，cd19)是特异性表达于人类b细胞表面的一种蛋白。cd19广泛表达在各个发育时期的b细胞表面并起到了重要的作用:cd19作为b细胞受体(bcellreceptor，bcr)共受体(co-receptor)，可以降低抗原介导的b细胞受体依赖性激活(antigenreceptor-dependentstimulation)所需的信号阈值；b细胞受体激活依赖于cd19胞内区磷酸化继而招募src激酶和pi3k激酶，从而彻底激活b细胞。完全成熟的b细胞称为浆细胞(plasmacell)，在完全成熟后浆细胞会丢失cd19抗原的表达。b细胞恶性增殖相关的血液肿瘤包括：b细胞急性淋巴细胞白血病(acutelymphoblasticleukemia，b-all)、b细胞慢性淋巴细胞白血病(chroniclymphoblasticleukemia，b-cll)以及b细胞淋巴瘤(blymphoma)。b细胞急性淋巴细胞白血病和b细胞淋巴瘤是恶性程度很高的癌症，其特征为外周血、骨髓伴随高程度的b细胞恶性增殖以及全身性b细胞实体瘤的形成，严重干扰患者的血液循环系统。现今，对于b细胞肿瘤的治疗主要包括小分子靶向药物比如伊马替尼(bcr-ablkinaseinhibitor)以及依鲁替尼(brutonkinaseinhibitor)、抗体药物比如利妥昔单抗(cd20antibody)以及骨髓移植(bonemarrowtransplantation)；在临床应用中，小分子药物以及抗体药物可以显著延长病人的生存期，为骨髓移植争取时间，但是部分病人会出现耐药性复发，至今临床上的药物对于耐药性复发的病人很难取得疗效。嵌合抗原受体t细胞疗法(cart)属于新型的治疗方式，以病人t细胞为基础在体外扩增并进行基因工程改造，cart细胞能够识别肿瘤细胞特异性抗原(例如cd19)，激活后可以特异性裂解肿瘤细胞达到杀伤肿瘤目的，其特点为治疗周期短、疗效较快、完全响应率高、能够为病人争取足够的时间等待骨髓移植，甚至彻底治愈。嵌合抗原受体组成为单链抗体(scfv)-铰链区(hingeregion)-跨膜区(transmembranedomain)-共刺激结构域(co-stimulatorydomain)-必要刺激结构域(essentialsignalingdomain)，其中对抗原具有高亲单链抗体为嵌合抗原受体(car)的最重要组成部分，scfv对于抗原的亲和力决定了cart细胞能否激活以及杀伤肿瘤细胞，因此对于抗原特异性scfv进行亲和力成熟的筛选尤其重要。fmc63-migg2a是上个世纪通过动物免疫获得抗cd19的鼠源抗体。fmc63scfv已经成功被应用于抗cd19car构造，比如norvatisctl019以及junotherapeuticsjcar015。ctl019以及jcar015在b细胞急性淋巴细胞白血病的临床试验取得了较好的结果，完全缓解(completeremission)病人比例>70％。技术实现要素：鉴于以上所述现有技术的缺点，本发明的目的在于提供一种抗cd19抗体及其制备方法和用途，用于解决现有技术中的问题。为实现上述目的及其他相关目的，本发明一方面提供一种抗cd19抗体，所述抗cd19抗体包括重链可变区和轻链可变区，所述抗cd19抗体具有如下技术特征中的一个或多个：<1>重链可变区的互补决定区包括氨基酸序列如seqidno.1所示的cdr-h1；<2>重链可变区的互补决定区包括氨基酸序列如seqidno.2所示的cdr-h2；<3>重链可变区的互补决定区包括氨基酸序列如seqidno.3所示的cdr-h3；<4>轻链可变区的互补决定区包括氨基酸序列如seqidno.4所示的cdr-l1；<5>轻链可变区的互补决定区包括氨基酸序列如seqidno.5、seqidno.6或seqidno.7所示的cdr-l2；<6>轻链可变区的互补决定区包括氨基酸序列如seqidno.8所示的cdr-l3。gwsledygvs(seqidno.1)viwgsettyynsalks(seqidno.2)hryyggsfdmdy(seqidno.3)rasqdiskyln(seqidno.4)qtsrlaw(seqidno.5)qtdrkqs(seqidno.6)htsrlhs(seqidno.7)qqgntlpyt(seqidno.8)cdr(互补决定区，complementaritydeterminingregion)通常指抗体中在空间结构上可以与抗原决定簇形成互补的区域。抗体中的可变性通常并不均匀地分布在整个抗体的可变区中，单克隆抗体的重链可变区和轻链可变区通常均具有3个超变区(hypervariableregion，hvr)，这些区域在空间结构上通常可以与抗原决定簇形成互补，所以超变区也被称为互补决定区(complementaritydeterminingregion，cdr)，即重链可变区通常包括三个互补决定区，即hcdr1、hcdr2和hcdr3，轻链可变区通常包括三个互补决定区，即lcdr1、lcdr2和lcdr3。在本发明某些实施方式中，所述抗cd19抗体的重链可变区的互补决定区包括氨基酸序列如seqidno.1所示的cdr-h1、氨基酸序列如seqidno.2所示的cdr-h2和氨基酸序列如seqidno.3所示的cdr-h3。在本发明某些实施方式中，所述抗cd19抗体的轻链可变区的互补决定区包括氨基酸序列如seqidno.4所示的cdr-l1、氨基酸序列如seqidno.5、seqidno.6或seqidno.7所示的cdr-l2和氨基酸序列如seqidno.8所示的cdr-l3。在本发明某些实施方式中，重链可变区的互补决定区包括氨基酸序列如seqidno.1所示的cdr-h1、氨基酸序列如seqidno.2所示的cdr-h2和氨基酸序列如seqidno.3所示的cdr-h3，轻链可变区的互补决定区包括氨基酸序列如seqidno.4所示的cdr-l1、氨基酸序列如seqidno.5、seqidno.6或seqidno.7所示的cdr-l2和氨基酸序列如seqidno.8所示的cdr-l3。在本发明某些实施方式中，所述抗cd19抗体为单克隆抗体。单克隆抗体通常指一个抗体的群体，该群体中所包括的抗体是基本相同的(除少数可能存在的天然发生的突变外)。单克隆抗体通常针对抗原上特定的决定簇。在本发明某些实施方式中，所述抗cd19抗体为单链抗体(singlechainfv,scfv)。单链抗体通常可以为包含抗体的vh(重链可变区)和vl(轻链可变区)的多肽链。通常来说，单链抗体还可以包括连接肽(linker)，连接肽通常位于vh和vl之间，以使scfv形成能与抗原结合的期望结构。例如，所述抗cd19抗体可以包括vh和vl，vh和vl之间可以设有连接肽，所述单链抗cd19抗体自n段至c端可以依次包括vh、连接肽和vl，所述抗cd19单链抗体自n段至c端也可以依次包括vl、连接肽和vh。所述连接肽可以是本领域中各种适用于形成scfv的连接肽，例如，所述连接肽可以是g4s3linker，所述g4s3linker的选择或设计可以参考文献michelsadelainetc，sciencetranslationalmedicine，2013；carlh.juneetc,sciencetranslationalmedicine,2015。在本发明某些实施方式中，所述抗cd19抗体衍生自cd19特异性的单克隆抗体fmc63(vh：y14283.1,vl:y14284.1)，其核苷酸序列如seqidno.9所示，氨基酸序列如seqidno.10所示。(seqidno.10，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)在本发明某些实施方式中，重链可变区和轻链可变区中还可以包括框架区，所述框架区可以位于互补决定区之间，也可以位于互补决定区的两端。在本发明一具体实施方式中，所述框架区的序列与fmc63的框架区序列一致，或为fmc63的框架区序列经过取代、缺失或者添加一个或多个(具体可以是1-50、1-30个、1-20个、1-10个、1-5个、或1-3个)氨基酸而得到的框架区序列，该框架区序列与fmc63的框架区序列可以具有80％、85％、90％、93％、95％、97％、或99％以上的同源性。在本发明某些实施方式中，所述抗cd19抗体的重链可变区的氨基酸序列包括：a)如seqidno.79-105其中之一所示的氨基酸序列；或b)与seqidno.79-105其中之一所示的氨基酸序列具有80％以上同源性、且具有a)所限定的氨基酸序列功能的氨基酸序列。具体的，所述b)中的氨基酸序列具体指：如seqidno.79-105其中之一所示的氨基酸序列经过取代、缺失或者添加一个或多个(具体可以是1-50、1-30个、1-20个、1-10个、1-5个、或1-3个)氨基酸而得到的，或者在n-末端和/或c-末端添加一个或多个(具体可以是1-50个、1-30个、1-20个、1-10个、1-5个、或1-3个)氨基酸而得到的，且具有如seqidno.79-105其中之一所示的氨基酸序列功能的氨基酸序列。所述b)中的氨基酸序列可与seqidno.79-105其中之一具有80％、85％、90％、93％、95％、97％、或99％以上的同源性。在本发明某些实施方式中，所述抗cd19抗体的轻链可变区的氨基酸序列包括：c)如seqidno.106-132其中之一所示的氨基酸序列；或d)与seqidno.106-132其中之一所示的氨基酸序列具有80％以上同源性、且具有c)所限定的氨基酸序列功能的氨基酸序列。具体的，所述d)中的氨基酸序列具体指：如seqidno.106-132其中之一所示的氨基酸序列经过取代、缺失或者添加一个或多个(具体可以是1-50、1-30个、1-20个、1-10个、1-5个、或1-3个)氨基酸而得到的，或者在n-末端和/或c-末端添加一个或多个(具体可以是1-50个、1-30个、1-20个、1-10个、1-5个、或1-3个)氨基酸而得到的，且具有如seqidno.106-132其中之一所示的氨基酸序列功能的氨基酸序列。所述b)中的氨基酸序列可与seqidno.106-132其中之一具有80％、85％、90％、93％、95％、97％、或99％以上的同源性。本发明另一方面提供一种分离的多核苷酸，编码所述抗cd19抗体的重链可变区和/或轻链可变区或全长氨基酸。本发明另一方面提供一种构建体，含有所述分离的多核苷酸。在本发明某些实施方式中，所述构建体由所述分离的多核苷酸插入到表达载体的多克隆位点构建而成。本发明中的表达载体通常指本领域熟知的各种市售表达载体，例如可以是细菌质粒、噬菌体、酵母质粒、植物细胞病毒、哺乳动物细胞病毒如腺病毒、逆转录病毒或其他载体。在本发明某些实施方式中，所述表达载体选自gv401表达载体(gv401为市售载体，供应商为吉凯基因)。本发明另一方面提供一种抗体的表达系统，所述表达系统含有所述构建体或基因组中整合有外源的所述多核苷酸。任何适用于表达载体进行表达的细胞都可以作为宿主细胞，例如，所述宿主细胞可以是原核细胞，如细菌细胞；或是低等真核细胞，如酵母细胞；或是高等真核细胞，如哺乳动物细胞。在本发明某些实施方式中，宿主细胞选自t细胞、nk细胞中的一种或多种的组合。本发明另一方面提供所述抗cd19抗体的制备方法，包括如下步骤：在适合表达所述抗体的条件下，培养所述的抗体的表达系统，从而表达出所述的抗体，纯化分离出所述的抗体。本发明中所用的宿主细胞均为现有技术，可通过商业途径直接获取，培养中所用的培养基亦为各种常规培养基，本领域技术人员可根据经验选择适用的培养基，在适于宿主细胞生长的条件下进行培养。当宿主细胞生长到适当的细胞密度后，用合适的方法(如温度转换或化学诱导)诱导选择的启动子，将细胞再培养一段时间。在上面的方法中的重组多肽可在细胞内、或在细胞膜上表达、或分泌到细胞外。如果需要，可利用其物理的、化学的和其它特性通过各种分离方法分离和纯化重组的蛋白。这些方法是本领域技术人员所熟知的。这些方法的例子包括但并不限于：常规的复性处理、用蛋白沉淀剂处理(盐析方法)、离心、渗透破菌、超处理、超离心、分子筛层析(凝胶过滤)、吸附层析、离子交换层析、高效液相层析(hplc)和其它各种液相层析技术及这些方法的结合。本发明另一方面提供所述抗cd19抗体在制备或筛选治疗药物中的用途、或制备诊断药物中的用途。所述治疗药物可以是以cd-19抗原为作用靶标，结合或作用于所述cd-19抗原，从而治疗和/或预防适应症的药物。在本发明某些实施方式中，所述治疗药物可以是肿瘤治疗药物。所述肿瘤治疗药物可以是以肿瘤细胞表面功能性表达的cd-19抗原为靶标，结合或作用于cd-19抗原，从而治疗和/或预防肿瘤的药物。所述肿瘤可以是急性淋巴型白血病、慢性淋巴型白血病、b淋巴瘤、或者其他与b细胞恶性增殖相关的肿瘤。在本发明某些实施方式中，所述治疗药物为嵌合抗原受体(car，chimericantigenreceptor)细胞治疗药物。所述嵌合抗原受体细胞治疗药物通常包括嵌合抗原受体细胞，所述嵌合抗原受体细胞可以是嵌合抗原受体t细胞、嵌合抗原受体nk细胞等，所述嵌合抗原受体t细胞通常包括t淋巴细胞，其还包括嵌合抗原受体。所述嵌合抗原受体nk细胞通常包括nk细胞，其还包括嵌合抗原受体。所述嵌合抗原受体包括跨膜域、胞内域和胞外域。在本发明某些实施方式中，所述胞外域包括所述抗cd19抗体，即所述嵌合抗原受体细胞可以在细胞表面表达所述抗cd-19抗体，从而可以引导该细胞对表达cd19抗原的细胞(例如，肿瘤细胞)进行作用的药物。所述对表达cd19抗原的细胞进行作用可以是杀死表达cd19抗原的细胞等。所以诊断药物具体指针对作用靶标cd19抗原，以cd19抗原作为生物标志物进行诊断的试剂。本发明另一方面提供一种分离的多肽，所述多肽包括跨膜域、胞内域和胞外域，所述胞外域包括所述抗cd19抗体。在本发明某些实施方式中，所述多肽为嵌合抗原受体。在本发明某些实施方式中，所述跨膜域可以包括cd8α(nm_001145873)、cd28(nm_006139)、dap10(nm_014266)等。在本发明某些实施方式中，所述胞内域可以包括共刺激结构域和/或信号结构域，例如，所述胞内域可以包括4-1bb(nm_001561)、cd28(nm_006139)、ox40(nm_003327)、icos(nm_012092)、cd3zeta(nm_198053)、dap10(nm_014266)等。在本发明某些实施方式中，所述多肽自n端至c端依次包括所述抗cd19单链抗体、跨膜域、胞内域。在本发明一些具体实施方式中，所述多肽自n端至c端依次包括所述抗cd19单链抗体、cd8α跨膜区、4-1bb共刺激结构域、cd3zeta信号结构域。在本发明一具体实施方式中，所述多肽自n端至c端依次包括所述抗cd19单链抗体、cd28跨膜区、cd28共刺激结构域、cd3zeta信号结构域。在本发明另一具体实施方式中，所述多肽自n端至c端依次包括所述抗cd19单链抗体、cd8α跨膜区、ox40共刺激结构域、cd3zeta信号结构域。在本发明另一具体实施方式中，所述多肽自n端至c端依次包括所述抗cd19单链抗体、cd8α跨膜区、icos共刺激结构域、cd3zeta信号结构域。在本发明另一具体实施方式中，所述多肽自n端至c端依次包括所述抗cd19单链抗体、cd8α跨膜区、4-1bb共刺激结构域、cd28共刺激结构域、cd3zeta。在本发明另一具体实施方式中，所述多肽自n端至c端依次包括所述抗cd19单链抗体、cd28跨膜区、cd28共刺激结构域、ox40共刺激结构域、cd3zeta信号结构域。本发明另一方面一种t淋巴细胞，所述t淋巴细胞含膜结合的所述多肽。在本发明某些实施方式中，所述多肽为嵌合抗原受体。所述t淋巴细胞通常可以表达所述多肽，其通常可以结合于cd19抗原，更具体可以通过包含所述抗cd19抗体的胞外域结合于cd19抗原，当所述多肽结合于所述cd19抗原时，所述t淋巴细胞通常可以活化和/或刺激从而得以增殖。在本发明某些实施方式中，所述胞外域包括所述抗cd19抗体，即所述嵌合抗原受体t细胞可以在t淋巴细胞表面表达所述抗cd-19抗体，从而可以引导t淋巴细胞对表达cd19抗原的细胞(例如，肿瘤细胞)进行作用的，所述作用可以是杀死表达cd19抗原的细胞等。本发明另一方面一种nk细胞，所述nk细胞含膜结合的所述多肽。在本发明某些实施方式中，所述多肽为嵌合抗原受体。所述nk细胞通常可以表达所述多肽，其通常可以结合于cd19抗原，更具体可以通过包含所述抗cd19抗体的胞外域结合于cd19抗原，当所述多肽结合于所述抗原时，所述nk细胞通常可以活化和/或刺激从而得以增殖。在本发明某些实施方式中，所述胞外域包括所述抗cd19抗体，即所述嵌合抗原受体nk细胞可以在nk细胞表面表达所述抗cd-19抗体，从而可以引导nk细胞对表达cd19抗原的细胞(例如，肿瘤细胞)进行作用的，所述作用可以是杀死表达cd19抗原的细胞等。本发明另一方面提供一种诊断试剂盒，包含诊断有效剂量的所述抗cd19抗体或其免疫偶联物。有效量通常指能够提供诊断效益的量。所以诊断试剂盒通常可以针对作用靶标cd19抗原，以cd19抗原作为生物标志物进行诊断。所述诊断试剂盒还可以包括抗cd19抗体的标记物，所述抗cd19抗体的标记物通常可以用于标记抗cd19抗体，可选用的标记物的种类包括但不限于荧光标记物、放射性标记物、酶标标记物、化学发光性标记物等中的一种或多种的组合。根据试剂盒的检测原理，所述试剂盒通常还可包含检测所需的一种或多种试剂。此外，所述试剂盒中还可根据需要包括：容器、对照物(阴性或阳性对照)、缓冲剂、助剂等，本领域技术人员可根据具体情况对其进行选择。本发明发明人利用噬菌体展示技术对fmc63scfv进行了亲和力成熟筛选，从而获得了高亲和力的对cd19的单链抗体，这些单链抗体能够竞争性阻断野生型fmc63migg2a对cd19的结合，从而可以提示这些单链抗体的抗原结合位点与fmc63migg2a一致。此外，本发明发明人进一步将高亲和力的单链抗体改造成嵌合抗原受体，例如利用表达抗cd19嵌合抗原受体的t细胞、nk细胞用于表达cd19的血液癌症(b-all，b-cll，b-lymphoma等)治疗，从而验证了突变后的嵌合抗原受体能够提高对肿瘤细胞的杀伤能力。附图说明图1-5显示为本发明实施例3实验结果示意图。图6-10显示为本发明实施例4实验结果示意图。图11-14显示为本发明实施例6实验结果示意图。图15显示为gv400载体结构示意图。图16显示为gv401载体结构示意图。图17-19显示为本发明实施例8实验结果示意图。图20显示为本发明实施例8实验结果汇总示意图。具体实施方式以下通过特定的具体实例说明本发明的实施方式，本领域技术人员可由本说明书所揭露的内容轻易地了解本发明的其他优点与功效。本发明还可以通过另外不同的具体实施方式加以实施或应用，本说明书中的各项细节也可以基于不同观点与应用，在没有背离本发明的精神下进行各种修饰或改变。在进一步描述本发明具体实施方式之前，应理解，本发明的保护范围不局限于下述特定的具体实施方案；还应当理解，本发明实施例中使用的术语是为了描述特定的具体实施方案，而不是为了限制本发明的保护范围；在本发明说明书和权利要求书中，除非文中另外明确指出，单数形式“一个”、“一”和“这个”包括复数形式。当实施例给出数值范围时，应理解，除非本发明另有说明，每个数值范围的两个端点以及两个端点之间任何一个数值均可选用。除非另外定义，本发明中使用的所有技术和科学术语与本
技术领域：
技术人员通常理解的意义相同。除实施例中使用的具体方法、设备、材料外，根据本
技术领域：
的技术人员对现有技术的掌握及本发明的记载，还可以使用与本发明实施例中所述的方法、设备、材料相似或等同的现有技术的任何方法、设备和材料来实现本发明。除非另外说明，本发明中所公开的实验方法、检测方法、制备方法均采用本
技术领域：
常规的分子生物学、生物化学、染色质结构和分析、分析化学、细胞培养、重组dna技术及相关领域的常规技术。这些技术在现有文献中已有完善说明，具体可参见sambrook等molecularcloning：alaboratorymanual，secondedition，coldspringharborlaboratorypress，1989andthirdedition，2001；ausubel等，currentprotocolsinmolecularbiology，johnwiley&sons，newyork，1987andperiodicupdates；theseriesmethodsinenzymology，academicpress，sandiego；wolffe，chromatinstructureandfunction，thirdedition，academicpress，sandiego，1998；methodsinenzymology，vol.304，chromatin(p.m.wassarmananda.p.wolffe，eds.)，academicpress，sandiego，1999；和methodsinmolecularbiology，vol.119，chromatinprotocols(p.b.becker，ed.)humanapress，totowa，1999等。实施例1fmc63scfv的重链(h)及轻链(l)的cdr1、cdr2、cdr3突变库的构建：fmc63重链及轻链cdr区域的选择基于可变区的氨基酸计数法(模板为单克隆抗体fmc63，vh：y14283.1,vl:y14284.1)，kabat计数(kabatnumberscheme.bioinf.org.uk)。以pcan-fmc63scfv质粒(由模板序列插入pcantab5e质粒(购自ge)的多克隆位点构建，模板序列为：vh：y14283.1,vl:y14284.1)为模板，利用随机引物pcr引入突变，引物如表2所示。获得的fmc63scfv的重链(h)及轻链(l)的cdr1、cdr2、cdr3突变库pcr产物分别命名为h1、h2、h3、l1、l2和l3。用sfii和noti对pcr产物及pcantab5e酶切回收后，经t4dna连接酶16℃连接过夜。连接产物电转至tg1感受态细胞，2xyt培养基重悬并于37℃复苏1h后，取菌液梯度稀释进行平板计数，得到各个突变库库容至少108，其余菌液全部涂布2xyt(ga)平板(葡萄糖2％，青霉素100ug/ml)。从上述突变库中分别随机挑取20个单克隆送测序，多样性均100％。表1实施例2噬菌体抗体库的淘选：加入20nmcd19-his-biotin抗原与噬菌体抗体库室温孵育2h，再将混合物转入链霉亲和素磁珠中室温共孵育15min。pbst-pbs洗去未结合的噬菌体，再加入胰酶37℃作用30min，从而洗脱下结合的噬菌体。将胰酶消化洗脱下的噬菌体感染4ml对数期的tg1菌体，37℃静置30min，取部分菌液梯度稀释进行平板计数，其余菌液全部涂布2xyt(ga)平板，用于下一轮的包装。包装后的噬菌体可用于下一轮的淘选，共进行4轮淘选富集，每轮淘选10倍稀释梯度降低抗原浓度，并逐轮增加pbst-pbs洗涤次数(四轮淘选的cd19-his-biotin抗原浓度分别为20nm、2nm、0.2nm和0.02nm，pbst-pbs洗涤次数分别为7，10，15，20次)。实施例3高亲和力scfv的筛选和鉴定：经过四轮的淘选后，随机挑取单克隆，iptg诱导后取上清进行elisa，elisa初步筛选后，挑取阳性信号至少2倍大于阴性信号的克隆送测序，分析测序结果，提取出富集较多的cdr区域对应的克隆。所述elisa初步筛选的具体步骤如下：挑取菌落克隆于4ml2xyt培养基中，37℃200rpm过夜培养，以1：100转接至500ml2xyt培养基中，37℃200rpm培养至对数生长期；加入m13k07辅助噬菌体(moi＝20)，37℃静置孵育30分钟，后于37℃200rpm孵育30分钟；离心去掉上清，菌体重悬至500ml2xyt/kan/amp抗性培养基中培养过夜；将培养过夜的上清用1/5体积peg/nacl溶液沉淀静置1小时后离心，将沉淀重悬于含30％丙三醇pbs溶液，4℃储存。将噬菌体溶液稀释2-10倍后测定a268，具体结果如表2所示(表2中，克隆号代表挑取的菌落克隆编号)。将avidin包被至96孔板，将biotin标记的cd19抗原结合至96孔，加入梯度稀释的噬菌体后孵育1小时(稀释前噬菌体原始浓度a268见表2)，用抗phage抗体(antim13phageantibody，ge公司购买)检测phage结合cd19，读取od450nm，结果如表3所示：ec50(a268)(ec50表示达到50％饱和信号所对应的噬菌体浓度，表3中ec50(a268)则表示ec50时所对应的a268的吸光度与噬菌体浓度成正比)越小，代表达到同样的饱和信号所需相应克隆稀释倍数越大，即相应克隆结合能力越强(a268通过nanodrop测定(thermofisher))；阳性对照为fmc63wt噬菌体，阴性对照为表达抗非cd19单链抗体噬菌体，使用方法同其他克隆一样)。具体结果如图1-5所示。表2表3克隆号ec50(a268)3h40.603c20.033c110.114e100.463f120.243h90.105a120.218c40.275g60.138c70.038b60.028c100.035a10.048b110.018d120.278a10.066d50.118c10.078f120.12阳性对照(fmc63wt)7.14阴性对照19.44实施例4fmc63scfv亲和力竞争性elisa筛选：将cdr突变重复较多的tg1克隆涂板，挑取单克隆，iptg诱导后纯化phage，纯化的phage经浓度测定后(a268＝5.4按照3倍浓度梯度稀释)，稀释梯度与3ug/mlfmc63migg2a竞争(孔板：将avidin包被至96孔板，将biotin标记的cd19抗原结合至96孔)，检测使用抗鼠fc二抗，加入底物(tmb)后孵育5mins，1m硫酸终止反应并读取od450nm，具体结果如表4、图6-10所示。比较不同克隆的ec50及emax(a268通过nanodrop测定(thermofisher)；阳性对照为fmc63wt噬菌体，阴性对照为表达抗非cd19单链抗体噬菌体，使用方法同其他克隆一样)。ec50(a268)(ec50表示能够达到最大抑制率emax50％对应的噬菌体浓度，表4中ec50(a268)则表示ec50时所对应的a268的吸光度)越小代表相应克隆的单链抗体竞争fmc63migg2a至50％结合能力越强；emax(最大抑制率，所有克隆噬菌体在吸光度5.4时能够竞争野生型抗体结合能力的百分比)越大代表相应克隆的单链抗体相同a268浓度下竞争能力越强。表4克隆号ec50(a268)emax(最大抑制率)3h41.9360％3c21.8745％3c111.2566％3h91.7526％4e10>5.410.6％3f12>5.49.5％5a122.9727％8c41.5845％5g61.9751％8c71.9270％8b60.8358％8c101.4656％5a10.9637％8b112.0459％8d122.0232％8a11.7148％6d51.8146％8c12.3648％8f12>5.46.7％阳性对照(fmc63wt)>5.420％阴性对照>5.419％根据以上结果，选取3h4，3c2，3c11，4e10，5a12，8c4，8b6进行组合突变测试(hcdr2富集序列不多，所以即使4e10竞争活性较差，但仍然选择)。实施例5fmc63scfv突变体kd/koff测定：将fmc63scfv突变体(各突变体对应的克隆号、及克隆号对应的scfv的核苷酸序列和氨基酸序列见下文，seqidno.25-51，seqidno.52-78)转接到pgciggh1(吉凯基因)(载体序列如seqidno.133所示)，构建可以表达分泌型fmc63scfv突变体higg1fc融合蛋白，利用真核表达系统表达；利用proteinacolumn进行纯化。将cd19抗原通过氨基偶联至ar2g芯片(fortiebio)，fmc63scfv突变体作为流动相，利用octetred96系统测定相应scfvkoffrate，测试结果表明(表5)，上述scfv突变体均对cd19抗原具有很好的亲和力。表5以上结果表明，各个克隆与fmc63wt拥有基本一致的koff值。实施例6高亲和力scfv的获得：实施例1-5中参考elisa结合、竞争及koff测试数据，选取如下的克隆通过pcr将相应cdr进行排列组合，组合结果如下(组合后获得的scfv对应的克隆号、及克隆号对应的scfv的核苷酸序列和氨基酸序列见下文，组合scfv序列cdr选自seqidno.25-51，seqidno.52-78)，通过大肠杆菌外周质(ppe)中含有scfv与cd19结合elisa实验进行筛选。fmc63突变库组合如下：表6重链comboscdr-h1cdr-h2cdr-h3combo53h44e103c2combo63h4fmc63wt3c2combo73h44e103c11combo83h4fmc63wt3c11轻链comboscdr-l1cdr-l2cdr-l3combo95a125g68b6combo105a12fmc63wt8b6combo138c45g68b6combo148c4fmc63wt8b6大肠杆菌外周质(ppe)制备：将cdr突变重复较多的tg1克隆涂板，挑取单克隆，iptg诱导后收集菌体重悬于tes(0.2mtris-hc1，0.5mmedta，0.5msucrose)中，再加入1/5×tes混匀，置冰上30min，经13000rpm离心获得上清，用0.22μm滤膜过滤后，4℃保存。将外周质按照一定比例进行稀释(2，4，8，16，32，64，128)加入cd19包被的96孔板中，进行elisa(实验方法参考实施例4)结果如图11-14所示。从实验结果可知，通过ppe稀释结合elisa实验combo5，combo7，combo9，combo10，combo14与cd19-his结合较弱，combo6，combo8，combo11，combo13与cd19结合力较强(在低稀释度时od450nm光吸收更高)，其中combo6结合量最高。克隆号：3h4(seqidno.52，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgwsledygvswirqsprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhyyyggsyamdywgqgtsvtvss(seqidno.79)diqmtqttsslsaslgdrvtiscrasqdiskylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgntlpytfgggtkleit(seqidno.106)克隆号：3c2(seqidno.53，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgvslpdygvswirqpprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhryyggsfdmdywgqgtsvtvss(seqidno.80)diqmtqttsslsaslgdrvtiscrasqdiskylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgntlpytfgggtkleit(seqidno.107)克隆号：3c11(seqidno.54，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgvslpdygvswirqpprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhwyyggsfamdywgqgtsvtvss(seqidno.81)diqmtqttsslsaslgdrvtiscrasqdiskylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgntlpytfgggtkleit(seqidno.108)克隆号：4e10(seqidno.55，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgvslpdygvswirqpprkglewlggiigsqttyylaglksrltiikdnsksqvflkmnslqtddtaiyycakhyyyggsyamdywgqgtsvtvss(seqidno.82)diqmtqttsslsaslgdrvtiscrasqdiskylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgntlpytfgggtkleit(seqidno.109)克隆号：3f12(seqidno.56，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgvslpdygvswirqpprkglewlgliqgswhtyygaalksrltiikdnsksqvflkmnslqtddtaiyycakhyyyggsyamdywgqgtsvtvss(seqidno.83)diqmtqttsslsaslgdrvtiscrasqdiskylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgntlpytfgggtkleit(seqidno.110)克隆号：8f12(seqidno.57，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgvslpdygvswirqpprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhyyyggsyamdywgqgtsvtvss(seqidno.84)diqmtqttsslsaslgdrvtiscrasqdiskylnwyqqkpdgtvklliyqtdrkqsgvpsrfsgsgsgtdysltisnleqediatyfcqqgntlpytfgggtkleit(seqidno.111)克隆号：3h9(seqidno.58，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgvslpdygvswirqpprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhyyyggsmamdywgqgtsvtvss(seqidno.85)diqmtqttsslsaslgdrvtiscrasqdiskylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgntlpytfgggtkleit(seqidno.112)克隆号：5a12(seqidno.59，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgvslpdygvswirqpprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhyyyggsyamdywgqgtsvtvss(seqidno.86)diqmtqttsslsaslgdrvtiscrasqpirrylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgntlpytfgggtkleit(seqidno.113)克隆号：8c4(seqidno.60，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgvslpdygvswirqsprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhyyyggsyamdywgqgtsvtvss(seqidno.87)diqmtqttsslsaslgdrvtiscrasmdikkylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgntlpytfgggtkleit(seqidno.114)克隆号：5g6(seqidno.61，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgvslpdygvswirqpprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhyyyggsyamdywgqgtsvtvss(seqidno.88)diqmtqttsslsaslgdrvtiscrasqdiskylnwyqqkpdgtvklliyqtsrlawgvpsrfsgsgsgtdysltisnleqediatyfcqqgntlpytfgggtkleit(seqidno.115)克隆号：8c7(seqidno.62，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgvslpdygvswirqpprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhyyyggsyamdywgqgtsvtvss(seqidno.89)diqmtqttsslsaslgdrvtiscrasqdiskylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgkaapltfgggtkleit(seqidno.116)克隆号：8b6(seqidno.63，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgvslpdygvswirqpprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhyyyggsyamdywgqgtsvtvss(seqidno.90)diqmtqttsslsaslgdrvtiscrasqdiskylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgkkapltfgggtkleit(seqidno.117)克隆号：8c10(seqidno.64，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgvslpdygvswirqpprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhyyyggsyamdywgqgtsvtvss(seqidno.91)diqmtqttsslsaslgdrvtiscrasqdiskylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgkkgpltfgggtkleit(seqidno.118)克隆号：5a1(seqidno.65，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgvslpdygvswirqpprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhyyyggsyamdywgqgtsvtvss(seqidno.92)diqmtqttsslsaslgdrvtiscrasqdiskylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgkrapltfgggtkleit(seqidno.119)克隆号：8b11(seqidno.66，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgvslpdygvswirqpprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhyyyggsyamdywgqgtsvtvss(seqidno.93)diqmtqttsslsaslgdrvtiscrasqdiskylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgkrgpltfgggtkleit(seqidno.120)克隆号：8d12(seqidno.67，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgvslpdygvswirqpprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhyyyggsyamdywgqgtsvtvss(seqidno.94)diqmtqttsslsaslgdrvtiscrasqdiskylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgktapltfgggtkleit(seqidno.121)克隆号：8a1(seqidno.68，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgvslpdygvswirqpprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhyyyggsyamdywgqgtsvtvss(seqidno.95)diqmtqttsslsaslgdrvtiscrasqdiskylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgktgpltfgggtkleit(seqidno.122)克隆号：6d5(seqidno.69，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgvslpdygvswirqpprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhyyyggsyamdywgqgtsvtvss(seqidno.96)diqmtqttsslsaslgdrvtiscrasqdiskylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgktlpftfgggtkleit(seqidno.123)克隆号：8c1(seqidno.70，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgvslpdygvswirqpprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhyyyggsyamdywgqgtsvtvss(seqidno.97)diqmtqttsslsaslgdrvtiscrasqdiskylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgnkapltfgggtkleit(seqidno.124)克隆号：combo-5(seqidno.71，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgwsledygvswirqsprkglewlggiigsqttyylaglksrltiikdnsksqvflkmnslqtddtaiyycakhryyggsfdmdywgqgtsvtvss(seqidno.98)diqmtqttsslsaslgdrvtiscrasqdiskylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgntlpytfgggtkleit(seqidno.125)克隆号：combo-6(seqidno.72，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgwsledygvswirqsprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhryyggsfdmdywgqgtsvtvss(seqidno.99)diqmtqttsslsaslgdrvtiscrasqdiskylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgntlpytfgggtkleit(seqidno.126)克隆号：combo-7(seqidno.73，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgwsledygvswirqsprkglewlggiigsqttyylaglksrltiikdnsksqvflkmnslqtddtaiyycakhwyyggsfamdywgqgtsvtvss(seqidno.100)diqmtqttsslsaslgdrvtiscrasqdiskylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgntlpytfgggtkleit(seqidno.127)克隆号：combo-8(seqidno.74，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgwsledygvswirqsprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhwyyggsfamdywgqgtsvtvss(seqidno.101)diqmtqttsslsaslgdrvtiscrasqdiskylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgntlpytfgggtkleit(seqidno.128)克隆号：combo-9(seqidno.75，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgvslpdygvswirqpprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhyyyggsyamdywgqgtsvtvss(seqidno.102)diqmtqttsslsaslgdrvtiscrasqpirrylnwyqqkpdgtvklliyqtsrlawgvpsrfsgsgsgtdysltisnleqediatyfcqqgkkapltfgggtkleit(seqidno.129)克隆号：combo-10(seqidno.76，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgvslpdygvswirqpprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhyyyggsyamdywgqgtsvtvss(seqidno.103)diqmtqttsslsaslgdrvtiscrasqpirrylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgkkapltfgggtkleit(seqidno.130)克隆号：combo-13(seqidno.77，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgvslpdygvswirqsprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhyyyggsyamdywgqgtsvtvss(seqidno.104)diqmtqttsslsaslgdrvtiscrasmdikkylnwyqqkpdgtvklliyqtsrlawgvpsrfsgsgsgtdysltisnleqediatyfcqqgkkapltfgggtkleit(seqidno.131)克隆号：combo-14(seqidno.78，其中粗体加下划线部分为连接肽，连接肽之前为重链可变区，连接肽之后为轻链可变区，下划线部分依次为cdr-h1、cdr-h2、cdr-h3、cdr-l1、cdr-l2、cdr-l3)evklqesgpglvapsqslsvtctvsgvslpdygvswirqsprkglewlgviwgsettyynsalksrltiikdnsksqvflkmnslqtddtaiyycakhyyyggsyamdywgqgtsvtvss(seqidno.105)diqmtqttsslsaslgdrvtiscrasmdikkylnwyqqkpdgtvklliyhtsrlhsgvpsrfsgsgsgtdysltisnleqediatyfcqqgkkapltfgggtkleit(seqidno.132)载体pgciggh1全序列：agtgggaattggctccggtgcccgtcagtgggcagagcgcacatcgcccacagtccccgagaagttggggggaggggtcggcaattgaaccggtgcctagagaaggtggcgcggggtaaactgggaaagtgatgtcgtgtactggctccgcctttttcccgagggtgggggagaaccgtatataagtgcagtagtcgccgtgaacgttctttttcgcaacgggtttgccgccagaacacaggtaagtgccgtgtgtggttcccgcgggcctggcctctttacgggttatggcccttgcgtgccttgaattacttccacctggctgcagtacgtgattcttgatcccgagcttcgggttggaagtgggtgggagagttcgaggccttgcgcttaaggagccccttcgcctcgtgcttgagttgaggcctggcctgggcgctggggccgccgcgtgcgaatctggtggcaccttcgcgcctgtctcgctgctttcgataagtctctagccatttaaaatttttgatgacctgctgcgacgctttttttctggcaagatagtcttgtaaatgcgggccaagatctgcacactggtatttcggtttttggggccgcgggcggcgacggggcccgtgcgtcccagcgcacatgttcggcgaggcggggcctgcgagcgcggccaccgagaatcggacgggggtagtctcaagctggccggcctgctctggtgcctggcctcgcgccgccgtgtatcgccccgccctgggcggcaaggctggcccggtcggcaccagttgcgtgagcggaaagatggccgcttcccggccctgctgcagggagctcaaaatggaggacgcggcgctcgggagagcgggcgggtgagtcacccacacaaaggaaaagggcctttccgtcctcagccgtcgcttcatgtgactccacggagtaccgggcgccgtccaggcacctcgattagttctcgagcttttggagtacgtcgtctttaggttggggggaggggttttatgcgatggagtttccccacactgagtgggtggagactgaagttaggccagcttggcacttgatgtaattctccttggaatttgccctttttgagtttggatcttggttcattctcaagcctcagacagtggttcaaagtttttttcttccatttcaggtgtcgtgaggaagatctctagaagctgggtaccttgtgcccgggcgccaccatggagtttgggctgagctggctttttcttgtcgcgattcttaagggtgtccagtgcgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacaacagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggatgagctgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaatgagcggccgctcgaggccggcaaggccggatccagacatgataagatacattgatgagtttggacaaaccacaactagaatgcagtgaaaaaaatgctttatttgtgaaatttgtgatgctattgctttatttgtaaccattataagctgcaataaacaagttaacaacaacaattgcattcattttatgtttcaggttcagggggaggtgtgggaggttttttaaagcaagtaaaacctctacaaatgtggtatggctgattatgatccggctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggatacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcagccatgaggtcgactctagaggatcgatccccgccccggacgaactaaacctgactacgacatctctgccccttcttcgcggggcagtgcatgtaatcccttcagttggttggtacaacttgccaactgggccctgttccacatgtgacacggggggggaccaaacacaaaggggttctctgactgtagttgacatccttataaatggatgtgcacatttgccaacactgagtggctttcatcctggagcagactttgcagtctgtggactgcaacacaacattgcctttatgtgtaactcttggctgaagctcttacaccaatgctgggggacatgtacctcccaggggcccaggaagactacgggaggctacaccaacgtcaatcagaggggcctgtgtagctaccgataagcggaccctcaagagggcattagcaatagtgtttataaggcccccttgttaaccctaaacgggtagcatatgcttcccgggtagtagtatatactatccagactaaccctaattcaatagcatatgttacccaacgggaagcatatgctatcgaattagggttagtaaaagggtcctaaggaacagcgatatctcccaccccatgagctgtcacggttttatttacatggggtcaggattccacgagggtagtgaaccattttagtcacaagggcagtggctgaagatcaaggagcgggcagtgaactctcctgaatcttcgcctgcttcttcattctccttcgtttagctaatagaataactgctgagttgtgaacagtaaggtgtatgtgaggtgctcgaaaacaaggtttcaggtgacgcccccagaataaaatttggacggggggttcagtggtggcattgtgctatgacaccaatataaccctcacaaaccccttgggcaataaatactagtgtaggaatgaaacattctgaatatctttaacaatagaaatccatggggtggggacaagccgtaaagactggatgtccatctcacacgaatttatggctatgggcaacacataatcctagtgcaatatgatactggggttattaagatgtgtcccaggcagggaccaagacaggtgaaccatgttgttacactctatttgtaacaaggggaaagagagtggacgccgacagcagcggactccactggttgtctctaacacccccgaaaattaaacggggctccacgccaatggggcccataaacaaagacaagtggccactcttttttttgaaattgtggagtgggggcacgcgtcagcccccacacgccgccctgcggttttggactgtaaaataagggtgtaataacttggctgattgtaaccccgctaaccactgcggtcaaaccacttgcccacaaaaccactaatggcaccccggggaatacctgcataagtaggtgggcgggccaagataggggcgcgattgctgcgatctggaggacaaattacacacacttgcgcctgagcgccaagcacagggttgttggtcctcatattcacgaggtcgctgagagcacggtgggctaatgttgccatgggtagcatatactacccaaatatctggatagcatatgctatcctaatctatatctgggtagcataggctatcctaatctatatctgggtagcatatgctatcctaatctatatctgggtagtatatgctatcctaatttatatctgggtagcataggctatcctaatctatatctgggtagcatatgctatcctaatctatatctgggtagtatatgctatcctaatctgtatccgggtagcatatgctatcctaatagagattagggtagtatatgctatcctaatttatatctgggtagcatatactacccaaatatctggatagcatatgctatcctaatctatatctgggtagcatatgctatcctaatctatatctgggtagcataggctatcctaatctatatctgggtagcatatgctatcctaatctatatctgggtagtatatgctatcctaatttatatctgggtagcataggctatcctaatctatatctgggtagcatatgctatcctaatctatatctgggtagtatatgctatcctaatctgtatccgggtagcatatgctatcctcatgcatatacagtcagcatatgatacccagtagtagagtgggagtgctatcctttgcatatgccgccacctcccaagggggcgtgaattttcgctgcttgtccttttcctgctgcttatcgatgataagctgtcaaacatgagaattcttgaagacgaaagggcctcgtgatacgcctatttttataggttaatgtcatgataataatggtttcttagacgtcaggtggcacttttcggggaaatgtgcgcggaacccctatttgtttatttttctaaatacattcaaatatgtatccgctcatgagacaataaccctgataaatgcttcaataatattgaaaaaggaagagtatgagtattcaacatttccgtgtcgcccttattcccttttttgcggcattttgccttcctgtttttgctcacccagaaacgctggtgaaagtaaaagatgctgaagatcagttgggtgcacgagtgggttacatcgaactggatctcaacagcggtaagatccttgagagttttcgccccgaagaacgttttccaatgatgagcacttttaaagttctgctatgtggcgcggtattatcccgtgttgacgccgggcaagagcaactcggtcgccgcatacactattctcagaatgacttggttgagtactcaccagtcacagaaaagcatcttacggatggcatgacagtaagagaattatgcagtgctgccataaccatgagtgataacactgcggccaacttacttctgacaacgatcggaggaccgaaggagctaaccgcttttttgcacaacatgggggatcatgtaactcgccttgatcgttgggaaccggagctgaatgaagccataccaaacgacgagcgtgacaccacgatgcctgcagcaatggcaacaacgttgcgcaaactattaactggcgaactacttactctagcttcccggcaacaattaatagactggatggaggcggataaagttgcaggaccacttctgcgctcggcccttccggctggctggtttattgctgataaatctggagccggtgagcgtgggtctcgcggtatcattgcagcactggggccagatggtaagccctcccgtatcgtagttatctacacgacggggagtcaggcaactatggatgaacgaaatagacagatcgctgagataggtgcctcactgattaagcattggtaactgtcagaccaagtttactcatatatactttagattgatttaaaacttcatttttaatttaaaaggatctaggtgaagatcctttttgataatctcatgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgttcttctagtgtagccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgaagctgtccctgatggtcgtcatctacctgcctggacagcatggcctgcaacgcgggcatcccgatgccgccggaagcgagaagaatcataatggggaaggccatccagcctcgcgtcgcgaacgccagcaagacgtagcccagcgcgtcggccccgagatgcgccgcgtgcggctgctggagatggcggacgcgatggatatgttctgccaagggttggtttgcgcattcacagttctccgcaagaattgattggctccaattcttggagtggtgaatccgttagcgaggtgccgccctgcttcatccccgtggcccgttgctcgcgtttgctggcggtgtcactggccccgtgggttagggacggggtcccccatggggaatggtttatggttcgtgggggttattattttgggcgttgcgtggggtcaggtccacgactggactgagcagacagacccatggtttttggatggcctgggcatggaccgcatgtactggcgcgacacgaacaccgggcgtctgtggctgccaaacacccccgacccccaaaaaccaccgcgcggatttctggcgtgccaagctagtcgaccaattctcatgtttgacagcttatcatcgcagatccgggcaacgttgttgccattgctgcaggcgcagaactggtaggtatggaagatct(seqidno.133)实施例7将高亲和力的scfv改造为car：构建scfv-bbz载体：参照seqidno.25-132所示scfv序列，按照cdr突变的氨基酸对fmc63-bbz嵌合抗原受体进行突变，具体为将scfv序列与cd8α-4-1bb-cd3zeta通过pcr方法连接，得到突变后的fmc63mutant-bbz系列嵌合抗原受体，通过标准分子生物学方法将scfv-bbz插入gv400载体(bamhi与ecori之间，吉凯基因，载体示意图如图15所示)中ef-1启动子与wpre结构之间，挑取克隆如下：3f12，5a12，5g6，8c4，8f12，8d12及combo6。构建scfv-bbz-p2a-egfp载体：将突变后得到的scfv-bbz突变体，通过标准分子生物学方法插入gv401载体(bamhi与ecori之间，吉凯基因，载体示意图如图16所示)中ef-1启动子与p2a结构(atnfsllkqagdveenpgp，seqidno.134)之间。实施例8fmc63mutant-bbz体外验证将各scfv序列构建获得的fmc63mutant-bbz包装成慢病毒(lentivirus)后，感染人类原代pbmc；感染后的t细胞在体外扩增和培养，通过细胞因子释放和杀伤实验评估fmc63mutation能否增强t细胞杀伤肿瘤细胞的功能，具体结果如下：感染效率检测：gv401穿梭载体包装为慢病毒后，通过感染人类t细胞，scfv-bbz-2a-egfp转录为mrna，经核糖体翻译，在2a序列(atnfsllkqagdveenpgp，seqidno.135)末尾甘氨酸和辅氨酸之间断裂形成两条独立的蛋白链，因此scfv-bbz与egfp蛋白表达摩尔数保持一致，通过流式细胞仪检测fitc通道可以检测egfp表达效率代表慢病毒感染效率。细胞因子释放实验：将人类pbmc用cd3和cd28抗体(okt3克隆和15e8克隆，miltenyibiotec)刺激激活24小时，将各scfv序列构建获得的fmc63mutant-bbz包装成慢病毒(lentivirus)后感染上述pbmc，感染表达抗cd19嵌合抗原受体，pbmc继续培养8-10天后，收集t细胞，并将t细胞和raji细胞(表达cd19)按照比例1：1混合培养于2％血清的rpmi1640培养基16小时将混合培养上清利用bdcytometricbeadarraykit测定细胞因子的释放。实验表明，感染获得的pbmc(各scfv序列构建获得的fmc63mutant-bbz包装成慢病毒感染获得，表达fmc63mutant-bbz嵌合抗原受体)与表达fmc63wt-bbz相比均具有很强的与raji细胞结合的能力并能够释放细胞因子。各克隆感染效率如下：表7克隆感染效率％fmc63wt26.9％3f1262.8％5a1226.6％5g658.5％8c455.9％8f1255.0％8d1265.7％combo615.6％各克隆细胞因子释放倍数变化如下：表8克隆il2倍数ifnγ倍数fmc63wt106126combo6146158表9克隆il2倍数ifnγ倍数fmc63wt46165a1247418d12373f12118f123215表10克隆il2倍数ifnγ倍数fmc63wt1231095g6871078c47398细胞因子释放实验结果表明，8d12及3f12克隆对肿瘤细胞刺激没有显著的细胞因子释放，提示8d12及3f12克隆突变不利于t细胞对表达cd19肿瘤细胞的结合。5a12克隆在与fmc63wt基本一致的感染效率下(26.6％vs26.9％)释放细胞因子il2和ifnγ水平较高。combo6在较低的感染效率下il2释放水平显著高于fmc63wt。提示5a12(cdrl1)，3h4(cdrh1)，3c2(cdrh3)突变有利于对于cd19结合，提高细胞因子释放水平。肿瘤杀伤实验：将上述制备获得的t细胞(pbmc继续培养8-10天后，收集t细胞)和raji细胞按照一定的e：t比例，例如30：1，10：1，3：1，1：1比例混合培养于2％血清的rpmi1640培养基中培养4小时，将培养上清与ldh底物(cytotox96non-radioactivecytotoxicityassaykit,promega)1：1体积混合，室温孵育30分钟后读取490nm光吸收。实验结果表明，感染获得的pbmc(各scfv序列构建获得的fmc63mutant-bbz包装成慢病毒感染获得)与fmc63wt-bbz相比均具有很强的肿瘤杀伤能力。细胞杀伤实验结果(图17-19)表明，3f12克隆无显著的对肿瘤细胞杀伤能力，提示3f12突变不利于结合cd19。8f12、5a12克隆与fmc63wt相比，对肿瘤细胞杀伤能力无显著差异。combo6克隆与fmc63wt相比，对肿瘤细胞杀伤有显著提高。汇总后的实验结果如图20所示，combo6克隆与fmc63wt相比，在e:t为10和3比例条件下，对肿瘤细胞杀伤显著提高，提示combo6组合突变有利于t细胞识别肿瘤细胞。实施例9fmc63mutant-bbz体外验证将各scfv序列构建获得的fmc63mutant-bbz包装成慢病毒(lentivirus)后，感染nk92细胞(培养条件：rpmi1640+20％fbs+200iu/mlhil2)；感染后的nk92细胞在体外扩增和培养，通过细胞因子释放和杀伤实验评估fmc63mutation能否增强nk92细胞杀伤肿瘤细胞的功能，具体结果如下：细胞因子释放实验：nk92细胞系培养在rpmi1640+20％fbs+200iu/mlhil2中；将fmc63mutant-bbz包装成慢病毒后感染nk92细胞，体外经过扩增和培养；将nk92细胞和raji细胞(表达cd19)按照比例1：1混合培养于2％血清的rpmi1640培养基16小时，将混合培养上清利用bdcytometricbeadarraykit测定细胞因子的释放。实验表明，感染获得的nk92(各scfv序列构建获得的fmc63mutant-bbz包装成慢病毒感染获得，表达fmc63mutant-bbz嵌合抗原受体)与表达fmc63wt-bbz相比均具有很强的与raji细胞结合的能力并能够释放细胞因子。肿瘤杀伤实验：将上述制备获得的nk92细胞和raji细胞按照一定的e：t比例，例如30：1，10：1，3：1，1：1比例混合培养于2％血清的rpmi1640培养基中培养4小时，将培养上清与ldh底物(cytotox96non-radioactivecytotoxicityassaykit,promega)1：1体积混合，室温孵育30分钟后读取490nm光吸收。实验结果表明，感染获得的nk92细胞(各scfv序列构建获得的fmc63mutant-bbz包装成慢病毒感染获得)与fmc63wt-bbz相比均具有很强的肿瘤杀伤能力。综上所述，本发明有效克服了现有技术中的种种缺点而具高度产业利用价值。上述实施例仅例示性说明本发明的原理及其功效，而非用于限制本发明。任何熟悉此技术的人士皆可在不违背本发明的精神及范畴下，对上述实施例进行修饰或改变。因此，举凡所属
技术领域：
中具有通常知识者在未脱离本发明所揭示的精神与技术思想下所完成的一切等效修饰或改变，仍应由本发明的权利要求所涵盖。序列表<110>上海吉倍生物技术有限公司<120>一种抗cd19抗体及其制备方法和用途<150>2016108048313<151>2016-09-06<150>2016108335750<151>2016-09-20<160>135<170>siposequencelisting1.0<210>1<211>10<212>prt<213>人工序列(artificialsequence)<400>1glytrpserleugluasptyrglyvalser1510<210>2<211>16<212>prt<213>人工序列(artificialsequence)<400>2valiletrpglysergluthrthrtyrtyrasnseralaleulysser151015<210>3<211>12<212>prt<213>人工序列(artificialsequence)<400>3hisargtyrtyrglyglyserpheaspmetasptyr1510<210>4<211>11<212>prt<213>人工序列(artificialsequence)<400>4argalaserglnaspileserlystyrleuasn1510<210>5<211>7<212>prt<213>人工序列(artificialsequence)<400>5glnthrserargleualatrp15<210>6<211>7<212>prt<213>人工序列(artificialsequence)<400>6glnthrasparglysglnser15<210>7<211>7<212>prt<213>人工序列(artificialsequence)<400>7histhrserargleuhisser15<210>8<211>9<212>prt<213>人工序列(artificialsequence)<400>8glnglnglyasnthrleuprotyrthr15<210>9<211>726<212>dna<213>人工序列(artificialsequence)<400>9gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattac300tacggtggtagctatgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggcggtggctcgggcggtggtgggtcgggtggcggcggatctgacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaatacgcttccgtacacgttcggaggggggaccaagctggag720atcaca726<210>10<211>242<212>prt<213>人工序列(artificialsequence)<400>10gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnaspileserlystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglyasnthrleuprotyrthrpheglyglyglythrlysleuglu225230235240ilethr<210>11<211>67<212>dna<213>人工序列(artificialsequence)<400>11tcacatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagc60ctccacg67<210>12<211>79<212>dna<213>人工序列(artificialsequence)<400>12agggtctggagtggctgggagtaatatggggtagtgaaaccacatactataattcagctc60tcaaatccagactgaccat79<210>13<211>72<212>dna<213>人工序列(artificialsequence)<400>13ccatttactactgtgccaaacattattactacggtggtagctatgctatggactactggg60gccaaggaacct72<210>14<211>72<212>dna<213>人工序列(artificialsequence)<400>14acagagtcaccatcagttgcagggcaagtcaggacattagtaaatatttaaattggtatc60agcagaaaccag72<210>15<211>61<212>dna<213>人工序列(artificialsequence)<400>15ctgttaaactcctgatctaccatacatcaagattacactcaggagtcccatcaaggttca60g61<210>16<211>64<212>dna<213>人工序列(artificialsequence)<400>16ttgccacttacttttgccaacagggtaatacgcttccgtacacgttcggaggggggacca60agct64<210>17<211>20<212>dna<213>人工序列(artificialsequence)<400>17ccctcatagttagcgtaacg20<210>18<211>24<212>dna<213>人工序列(artificialsequence)<400>18agcggataacaatttcacacagga24<210>19<211>20<212>dna<213>人工序列(artificialsequence)<400>19ccctgagacagtgcatgtga20<210>20<211>20<212>dna<213>人工序列(artificialsequence)<400>20tcccagccactccagaccct20<210>21<211>20<212>dna<213>人工序列(artificialsequence)<400>21tttggcacagtagtaaatgg20<210>22<211>20<212>dna<213>人工序列(artificialsequence)<400>22gcaactgatggtgactctgt20<210>23<211>20<212>dna<213>人工序列(artificialsequence)<400>23gtagatcaggagtttaacag20<210>24<211>20<212>dna<213>人工序列(artificialsequence)<400>24ttggcaaaagtaagtggcaa20<210>25<211>726<212>dna<213>人工序列(artificialsequence)<400>25gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcagggtggtcattagaggactatggtgtaagctggattcgccagtct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattac300tacggtggtagctatgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaatacgcttccgtacacgttcggaggggggaccaagctggag720atcaca726<210>26<211>726<212>dna<213>人工序列(artificialsequence)<400>26gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacatcggtac300tacggtggtagctttgatatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaatacgcttccgtacacgttcggaggggggaccaagctggag720atcaca726<210>27<211>726<212>dna<213>人工序列(artificialsequence)<400>27gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattggtac300tacggtggtagctttgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaatacgcttccgtacacgttcggaggggggaccaagctggag720atcaca726<210>28<211>726<212>dna<213>人工序列(artificialsequence)<400>28gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcct120ccacgaaagggtctggagtggctgggagggataattggtagtcagaccacatactatctg180gctgggctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattac300tacggtggtagctatgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaatacgcttccgtacacgttcggaggggggaccaagctggag720atcaca726<210>29<211>726<212>dna<213>人工序列(artificialsequence)<400>29gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcct120ccacgaaagggtctggagtggctgggactgatacagggtagttggcatacatactatggt180gcggctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattac300tacggtggtagctatgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaatacgcttccgtacacgttcggaggggggaccaagctggag720atcaca726<210>30<211>726<212>dna<213>人工序列(artificialsequence)<400>30gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattac300tacggtggtagctatgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccagacagatagaaagcagtcgggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaatacgcttccgtacacgttcggaggggggaccaagctggag720atcaca726<210>31<211>726<212>dna<213>人工序列(artificialsequence)<400>31gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattat300tacggtggtagcatggctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaatacgcttccgtacacgttcggaggggggaccaagctggag720atcaca726<210>32<211>726<212>dna<213>人工序列(artificialsequence)<400>32gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattac300tacggtggtagctatgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcagcctattcggaggtatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaatacgcttccgtacacgttcggaggggggaccaagctggag720atcaca726<210>33<211>726<212>dna<213>人工序列(artificialsequence)<400>33gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagtct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattac300tacggtggtagctatgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtatggacattaagaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaatacgcttccgtacacgttcggaggggggaccaagctggag720atcaca726<210>34<211>726<212>dna<213>人工序列(artificialsequence)<400>34gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattac300tacggtggtagctatgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccagacatcaagattagcctggggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaatacgcttccgtacacgttcggaggggggaccaagctggag720atcaca726<210>35<211>726<212>dna<213>人工序列(artificialsequence)<400>35gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattac300tacggtggtagctatgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaaggcggctccgttgacgttcggaggggggaccaagctggag720atcaca726<210>36<211>726<212>dna<213>人工序列(artificialsequence)<400>36gaggtgaaactgcaggagtcaggacctggcctggtggcgccatcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattac300tacggtggtagctatgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacaggggaagaaggcgccgttgacgttcggaggggggaccaagctggag720atcaca726<210>37<211>726<212>dna<213>人工序列(artificialsequence)<400>37gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattac300tacggtggtagctatgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaagaaggggccgcttacgttcggaggggggaccaagctggag720atcaca726<210>38<211>726<212>dna<213>人工序列(artificialsequence)<400>38gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattac300tacggtggtagctatgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaagagggcgccgttgacgttcggaggggggaccaagctggag720atcaca726<210>39<211>726<212>dna<213>人工序列(artificialsequence)<400>39gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattac300tacggtggtagctatgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccgtcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaagagggggccgttgactttcggaggggggaccaagctggag720atcaca726<210>40<211>726<212>dna<213>人工序列(artificialsequence)<400>40gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattac300tacggtggtagctatgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaagacggctccgttgacgttcggaggggggaccaagctggag720atcaca726<210>41<211>726<212>dna<213>人工序列(artificialsequence)<400>41gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattac300tacggtggtagctatgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaagacggggccgttgacgttcggaggggggaccaagctggag720atcaca726<210>42<211>726<212>dna<213>人工序列(artificialsequence)<400>42gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattac300tacggtggtagctatgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaagacgcttccgtttacgttcggaggggggaccaagctggag720atcaca726<210>43<211>726<212>dna<213>人工序列(artificialsequence)<400>43gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattac300tacggtggtagctatgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaataaggcgccgctgacgttcggaggggggaccaagctggag720atcaca726<210>44<211>726<212>dna<213>人工序列(artificialsequence)<400>44gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcagggtggtcattagaggactatggtgtaagctggattcgccagtct120ccacgaaagggtctggagtggctgggagggataattggtagtcagaccacatactatctg180gctgggctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacatcggtac300tacggtggtagctttgatatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaatacgcttccgtacacgttcggaggggggaccaagctggag720atcaca726<210>45<211>726<212>dna<213>人工序列(artificialsequence)<400>45gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcagggtggtcattagaggactatggtgtaagctggattcgccagtct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacatcggtac300tacggtggtagctttgatatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaatacgcttccgtacacgttcggaggggggaccaagctggag720atcaca726<210>46<211>726<212>dna<213>人工序列(artificialsequence)<400>46gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcagggtggtcattagaggactatggtgtaagctggattcgccagtct120ccacgaaagggtctggagtggctgggagggataattggtagtcagaccacatactatctg180gctgggctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattggtac300tacggtggtagctttgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaatacgcttccgtacacgttcggaggggggaccaagctggag720atcaca726<210>47<211>726<212>dna<213>人工序列(artificialsequence)<400>47gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcagggtggtcattagaggactatggtgtaagctggattcgccagtct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattggtac300tacggtggtagctttgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacagggtaatacgcttccgtacacgttcggaggggggaccaagctggag720atcaca726<210>48<211>726<212>dna<213>人工序列(artificialsequence)<400>48gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattac300tacggtggtagctatgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcagcctattcggaggtatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccagacatcaagattagcctggggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacaggggaagaaggcgccgttgacgttcggaggggggaccaagctggag720atcaca726<210>49<211>726<212>dna<213>人工序列(artificialsequence)<400>49gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattac300tacggtggtagctatgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtcagcctattcggaggtatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacaggggaagaaggcgccgttgacgttcggaggggggaccaagctggag720atcaca726<210>50<211>726<212>dna<213>人工序列(artificialsequence)<400>50gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagtct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattac300tacggtggtagctatgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtatggacattaagaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccagacatcaagattagcctggggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacaggggaagaaggcgccgttgacgttcggaggggggaccaagctggag720atcaca726<210>51<211>726<212>dna<213>人工序列(artificialsequence)<400>51gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtc60acatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagtct120ccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat180tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttctta240aaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattac300tacggtggtagctatgctatggactactggggccaaggaacctcagtcaccgtctcctca360ggtggaggcggttcaggcggaggtggctctggcggtggcggatcggacatccagatgaca420cagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggca480agtatggacattaagaaatatttaaattggtatcagcagaaaccagatggaactgttaaa540ctcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagt600gggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccact660tacttttgccaacaggggaagaaggcgccgttgacgttcggaggggggaccaagctggag720atcaca726<210>52<211>242<212>prt<213>人工序列(artificialsequence)<400>52gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglytrpserleugluasptyr202530glyvalsertrpileargglnserproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnaspileserlystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglyasnthrleuprotyrthrpheglyglyglythrlysleuglu225230235240ilethr<210>53<211>242<212>prt<213>人工序列(artificialsequence)<400>53gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshisargtyrtyrglyglyserpheaspmetasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnaspileserlystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglyasnthrleuprotyrthrpheglyglyglythrlysleuglu225230235240ilethr<210>54<211>242<212>prt<213>人工序列(artificialsequence)<400>54gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistrptyrtyrglyglyserphealametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnaspileserlystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglyasnthrleuprotyrthrpheglyglyglythrlysleuglu225230235240ilethr<210>55<211>242<212>prt<213>人工序列(artificialsequence)<400>55gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyglyileileglyserglnthrthrtyrtyrleualaglyleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnaspileserlystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglyasnthrleuprotyrthrpheglyglyglythrlysleuglu225230235240ilethr<210>56<211>242<212>prt<213>人工序列(artificialsequence)<400>56gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyleuileglnglysertrphisthrtyrtyrglyalaalaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnaspileserlystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglyasnthrleuprotyrthrpheglyglyglythrlysleuglu225230235240ilethr<210>57<211>242<212>prt<213>人工序列(artificialsequence)<400>57gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnaspileserlystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrglnthrasparglysglnsergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglyasnthrleuprotyrthrpheglyglyglythrlysleuglu225230235240ilethr<210>58<211>242<212>prt<213>人工序列(artificialsequence)<400>58gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysermetalametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnaspileserlystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglyasnthrleuprotyrthrpheglyglyglythrlysleuglu225230235240ilethr<210>59<211>242<212>prt<213>人工序列(artificialsequence)<400>59gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnproileargargtyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglyasnthrleuprotyrthrpheglyglyglythrlysleuglu225230235240ilethr<210>60<211>242<212>prt<213>人工序列(artificialsequence)<400>60gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnserproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160sermetaspilelyslystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglyasnthrleuprotyrthrpheglyglyglythrlysleuglu225230235240ilethr<210>61<211>242<212>prt<213>人工序列(artificialsequence)<400>61gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnaspileserlystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrglnthrserargleualatrpgly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglyasnthrleuprotyrthrpheglyglyglythrlysleuglu225230235240ilethr<210>62<211>242<212>prt<213>人工序列(artificialsequence)<400>62gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnaspileserlystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglylysalaalaproleuthrpheglyglyglythrlysleuglu225230235240ilethr<210>63<211>242<212>prt<213>人工序列(artificialsequence)<400>63gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnaspileserlystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglylyslysalaproleuthrpheglyglyglythrlysleuglu225230235240ilethr<210>64<211>242<212>prt<213>人工序列(artificialsequence)<400>64gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnaspileserlystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglylyslysglyproleuthrpheglyglyglythrlysleuglu225230235240ilethr<210>65<211>242<212>prt<213>人工序列(artificialsequence)<400>65gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnaspileserlystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglylysargalaproleuthrpheglyglyglythrlysleuglu225230235240ilethr<210>66<211>242<212>prt<213>人工序列(artificialsequence)<400>66gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnaspileserlystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglylysargglyproleuthrpheglyglyglythrlysleuglu225230235240ilethr<210>67<211>242<212>prt<213>人工序列(artificialsequence)<400>67gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnaspileserlystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglylysthralaproleuthrpheglyglyglythrlysleuglu225230235240ilethr<210>68<211>242<212>prt<213>人工序列(artificialsequence)<400>68gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnaspileserlystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglylysthrglyproleuthrpheglyglyglythrlysleuglu225230235240ilethr<210>69<211>242<212>prt<213>人工序列(artificialsequence)<400>69gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnaspileserlystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglylysthrleuprophethrpheglyglyglythrlysleuglu225230235240ilethr<210>70<211>242<212>prt<213>人工序列(artificialsequence)<400>70gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnaspileserlystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglyasnlysalaproleuthrpheglyglyglythrlysleuglu225230235240ilethr<210>71<211>242<212>prt<213>人工序列(artificialsequence)<400>71gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglytrpserleugluasptyr202530glyvalsertrpileargglnserproarglysglyleuglutrpleu354045glyglyileileglyserglnthrthrtyrtyrleualaglyleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshisargtyrtyrglyglyserpheaspmetasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnaspileserlystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglyasnthrleuprotyrthrpheglyglyglythrlysleuglu225230235240ilethr<210>72<211>242<212>prt<213>人工序列(artificialsequence)<400>72gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglytrpserleugluasptyr202530glyvalsertrpileargglnserproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshisargtyrtyrglyglyserpheaspmetasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnaspileserlystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglyasnthrleuprotyrthrpheglyglyglythrlysleuglu225230235240ilethr<210>73<211>242<212>prt<213>人工序列(artificialsequence)<400>73gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglytrpserleugluasptyr202530glyvalsertrpileargglnserproarglysglyleuglutrpleu354045glyglyileileglyserglnthrthrtyrtyrleualaglyleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistrptyrtyrglyglyserphealametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnaspileserlystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglyasnthrleuprotyrthrpheglyglyglythrlysleuglu225230235240ilethr<210>74<211>242<212>prt<213>人工序列(artificialsequence)<400>74gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglytrpserleugluasptyr202530glyvalsertrpileargglnserproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistrptyrtyrglyglyserphealametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnaspileserlystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglyasnthrleuprotyrthrpheglyglyglythrlysleuglu225230235240ilethr<210>75<211>242<212>prt<213>人工序列(artificialsequence)<400>75gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnproileargargtyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrglnthrserargleualatrpgly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglylyslysalaproleuthrpheglyglyglythrlysleuglu225230235240ilethr<210>76<211>242<212>prt<213>人工序列(artificialsequence)<400>76gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160serglnproileargargtyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglylyslysalaproleuthrpheglyglyglythrlysleuglu225230235240ilethr<210>77<211>242<212>prt<213>人工序列(artificialsequence)<400>77gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnserproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160sermetaspilelyslystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrglnthrserargleualatrpgly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglylyslysalaproleuthrpheglyglyglythrlysleuglu225230235240ilethr<210>78<211>242<212>prt<213>人工序列(artificialsequence)<400>78gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnserproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserserglyglyglyglyserglyglygly115120125glyserglyglyglyglyseraspileglnmetthrglnthrthrser130135140serleuseralaserleuglyaspargvalthrilesercysargala145150155160sermetaspilelyslystyrleuasntrptyrglnglnlysproasp165170175glythrvallysleuleuiletyrhisthrserargleuhissergly180185190valproserargpheserglyserglyserglythrasptyrserleu195200205thrileserasnleugluglngluaspilealathrtyrphecysgln210215220glnglylyslysalaproleuthrpheglyglyglythrlysleuglu225230235240ilethr<210>79<211>120<212>prt<213>人工序列(artificialsequence)<400>79gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglytrpserleugluasptyr202530glyvalsertrpileargglnserproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>80<211>120<212>prt<213>人工序列(artificialsequence)<400>80gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshisargtyrtyrglyglyserpheaspmetasptyrtrpglygln100105110glythrservalthrvalserser115120<210>81<211>120<212>prt<213>人工序列(artificialsequence)<400>81gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistrptyrtyrglyglyserphealametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>82<211>120<212>prt<213>人工序列(artificialsequence)<400>82gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyglyileileglyserglnthrthrtyrtyrleualaglyleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>83<211>120<212>prt<213>人工序列(artificialsequence)<400>83gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyleuileglnglysertrphisthrtyrtyrglyalaalaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>84<211>120<212>prt<213>人工序列(artificialsequence)<400>84gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>85<211>120<212>prt<213>人工序列(artificialsequence)<400>85gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysermetalametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>86<211>120<212>prt<213>人工序列(artificialsequence)<400>86gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>87<211>120<212>prt<213>人工序列(artificialsequence)<400>87gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnserproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>88<211>120<212>prt<213>人工序列(artificialsequence)<400>88gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>89<211>120<212>prt<213>人工序列(artificialsequence)<400>89gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>90<211>120<212>prt<213>人工序列(artificialsequence)<400>90gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>91<211>120<212>prt<213>人工序列(artificialsequence)<400>91gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>92<211>120<212>prt<213>人工序列(artificialsequence)<400>92gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>93<211>120<212>prt<213>人工序列(artificialsequence)<400>93gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>94<211>120<212>prt<213>人工序列(artificialsequence)<400>94gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>95<211>120<212>prt<213>人工序列(artificialsequence)<400>95gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>96<211>120<212>prt<213>人工序列(artificialsequence)<400>96gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>97<211>120<212>prt<213>人工序列(artificialsequence)<400>97gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>98<211>120<212>prt<213>人工序列(artificialsequence)<400>98gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglytrpserleugluasptyr202530glyvalsertrpileargglnserproarglysglyleuglutrpleu354045glyglyileileglyserglnthrthrtyrtyrleualaglyleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshisargtyrtyrglyglyserpheaspmetasptyrtrpglygln100105110glythrservalthrvalserser115120<210>99<211>120<212>prt<213>人工序列(artificialsequence)<400>99gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglytrpserleugluasptyr202530glyvalsertrpileargglnserproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshisargtyrtyrglyglyserpheaspmetasptyrtrpglygln100105110glythrservalthrvalserser115120<210>100<211>120<212>prt<213>人工序列(artificialsequence)<400>100gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglytrpserleugluasptyr202530glyvalsertrpileargglnserproarglysglyleuglutrpleu354045glyglyileileglyserglnthrthrtyrtyrleualaglyleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistrptyrtyrglyglyserphealametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>101<211>120<212>prt<213>人工序列(artificialsequence)<400>101gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglytrpserleugluasptyr202530glyvalsertrpileargglnserproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistrptyrtyrglyglyserphealametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>102<211>120<212>prt<213>人工序列(artificialsequence)<400>102gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>103<211>120<212>prt<213>人工序列(artificialsequence)<400>103gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnproproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>104<211>120<212>prt<213>人工序列(artificialsequence)<400>104gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnserproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>105<211>120<212>prt<213>人工序列(artificialsequence)<400>105gluvallysleuglngluserglyproglyleuvalalaprosergln151015serleuservalthrcysthrvalserglyvalserleuproasptyr202530glyvalsertrpileargglnserproarglysglyleuglutrpleu354045glyvaliletrpglysergluthrthrtyrtyrasnseralaleulys505560serargleuthrileilelysaspasnserlysserglnvalpheleu65707580lysmetasnserleuglnthraspaspthralailetyrtyrcysala859095lyshistyrtyrtyrglyglysertyralametasptyrtrpglygln100105110glythrservalthrvalserser115120<210>106<211>107<212>prt<213>人工序列(artificialsequence)<400>106aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnaspileserlystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglyasnthrleuprotyr859095thrpheglyglyglythrlysleugluilethr100105<210>107<211>107<212>prt<213>人工序列(artificialsequence)<400>107aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnaspileserlystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglyasnthrleuprotyr859095thrpheglyglyglythrlysleugluilethr100105<210>108<211>107<212>prt<213>人工序列(artificialsequence)<400>108aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnaspileserlystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglyasnthrleuprotyr859095thrpheglyglyglythrlysleugluilethr100105<210>109<211>107<212>prt<213>人工序列(artificialsequence)<400>109aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnaspileserlystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglyasnthrleuprotyr859095thrpheglyglyglythrlysleugluilethr100105<210>110<211>107<212>prt<213>人工序列(artificialsequence)<400>110aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnaspileserlystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglyasnthrleuprotyr859095thrpheglyglyglythrlysleugluilethr100105<210>111<211>107<212>prt<213>人工序列(artificialsequence)<400>111aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnaspileserlystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrglnthrasparglysglnserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglyasnthrleuprotyr859095thrpheglyglyglythrlysleugluilethr100105<210>112<211>107<212>prt<213>人工序列(artificialsequence)<400>112aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnaspileserlystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglyasnthrleuprotyr859095thrpheglyglyglythrlysleugluilethr100105<210>113<211>107<212>prt<213>人工序列(artificialsequence)<400>113aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnproileargargtyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglyasnthrleuprotyr859095thrpheglyglyglythrlysleugluilethr100105<210>114<211>107<212>prt<213>人工序列(artificialsequence)<400>114aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalasermetaspilelyslystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglyasnthrleuprotyr859095thrpheglyglyglythrlysleugluilethr100105<210>115<211>107<212>prt<213>人工序列(artificialsequence)<400>115aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnaspileserlystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrglnthrserargleualatrpglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglyasnthrleuprotyr859095thrpheglyglyglythrlysleugluilethr100105<210>116<211>107<212>prt<213>人工序列(artificialsequence)<400>116aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnaspileserlystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglylysalaalaproleu859095thrpheglyglyglythrlysleugluilethr100105<210>117<211>107<212>prt<213>人工序列(artificialsequence)<400>117aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnaspileserlystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglylyslysalaproleu859095thrpheglyglyglythrlysleugluilethr100105<210>118<211>107<212>prt<213>人工序列(artificialsequence)<400>118aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnaspileserlystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglylyslysglyproleu859095thrpheglyglyglythrlysleugluilethr100105<210>119<211>107<212>prt<213>人工序列(artificialsequence)<400>119aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnaspileserlystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglylysargalaproleu859095thrpheglyglyglythrlysleugluilethr100105<210>120<211>107<212>prt<213>人工序列(artificialsequence)<400>120aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnaspileserlystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglylysargglyproleu859095thrpheglyglyglythrlysleugluilethr100105<210>121<211>107<212>prt<213>人工序列(artificialsequence)<400>121aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnaspileserlystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglylysthralaproleu859095thrpheglyglyglythrlysleugluilethr100105<210>122<211>107<212>prt<213>人工序列(artificialsequence)<400>122aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnaspileserlystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglylysthrglyproleu859095thrpheglyglyglythrlysleugluilethr100105<210>123<211>107<212>prt<213>人工序列(artificialsequence)<400>123aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnaspileserlystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglylysthrleuprophe859095thrpheglyglyglythrlysleugluilethr100105<210>124<211>107<212>prt<213>人工序列(artificialsequence)<400>124aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnaspileserlystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglyasnlysalaproleu859095thrpheglyglyglythrlysleugluilethr100105<210>125<211>107<212>prt<213>人工序列(artificialsequence)<400>125aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnaspileserlystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglyasnthrleuprotyr859095thrpheglyglyglythrlysleugluilethr100105<210>126<211>107<212>prt<213>人工序列(artificialsequence)<400>126aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnaspileserlystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglyasnthrleuprotyr859095thrpheglyglyglythrlysleugluilethr100105<210>127<211>107<212>prt<213>人工序列(artificialsequence)<400>127aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnaspileserlystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglyasnthrleuprotyr859095thrpheglyglyglythrlysleugluilethr100105<210>128<211>107<212>prt<213>人工序列(artificialsequence)<400>128aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnaspileserlystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglyasnthrleuprotyr859095thrpheglyglyglythrlysleugluilethr100105<210>129<211>107<212>prt<213>人工序列(artificialsequence)<400>129aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnproileargargtyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrglnthrserargleualatrpglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglylyslysalaproleu859095thrpheglyglyglythrlysleugluilethr100105<210>130<211>107<212>prt<213>人工序列(artificialsequence)<400>130aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalaserglnproileargargtyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglylyslysalaproleu859095thrpheglyglyglythrlysleugluilethr100105<210>131<211>107<212>prt<213>人工序列(artificialsequence)<400>131aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalasermetaspilelyslystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrglnthrserargleualatrpglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglylyslysalaproleu859095thrpheglyglyglythrlysleugluilethr100105<210>132<211>107<212>prt<213>人工序列(artificialsequence)<400>132aspileglnmetthrglnthrthrserserleuseralaserleugly151015aspargvalthrilesercysargalasermetaspilelyslystyr202530leuasntrptyrglnglnlysproaspglythrvallysleuleuile354045tyrhisthrserargleuhisserglyvalproserargphesergly505560serglyserglythrasptyrserleuthrileserasnleuglugln65707580gluaspilealathrtyrphecysglnglnglylyslysalaproleu859095thrpheglyglyglythrlysleugluilethr100105<210>133<211>7252<212>dna<213>人工序列(artificialsequence)<400>133agtgggaattggctccggtgcccgtcagtgggcagagcgcacatcgcccacagtccccga60gaagttggggggaggggtcggcaattgaaccggtgcctagagaaggtggcgcggggtaaa120ctgggaaagtgatgtcgtgtactggctccgcctttttcccgagggtgggggagaaccgta180tataagtgcagtagtcgccgtgaacgttctttttcgcaacgggtttgccgccagaacaca240ggtaagtgccgtgtgtggttcccgcgggcctggcctctttacgggttatggcccttgcgt300gccttgaattacttccacctggctgcagtacgtgattcttgatcccgagcttcgggttgg360aagtgggtgggagagttcgaggccttgcgcttaaggagccccttcgcctcgtgcttgagt420tgaggcctggcctgggcgctggggccgccgcgtgcgaatctggtggcaccttcgcgcctg480tctcgctgctttcgataagtctctagccatttaaaatttttgatgacctgctgcgacgct540ttttttctggcaagatagtcttgtaaatgcgggccaagatctgcacactggtatttcggt600ttttggggccgcgggcggcgacggggcccgtgcgtcccagcgcacatgttcggcgaggcg660gggcctgcgagcgcggccaccgagaatcggacgggggtagtctcaagctggccggcctgc720tctggtgcctggcctcgcgccgccgtgtatcgccccgccctgggcggcaaggctggcccg780gtcggcaccagttgcgtgagcggaaagatggccgcttcccggccctgctgcagggagctc840aaaatggaggacgcggcgctcgggagagcgggcgggtgagtcacccacacaaaggaaaag900ggcctttccgtcctcagccgtcgcttcatgtgactccacggagtaccgggcgccgtccag960gcacctcgattagttctcgagcttttggagtacgtcgtctttaggttggggggaggggtt1020ttatgcgatggagtttccccacactgagtgggtggagactgaagttaggccagcttggca1080cttgatgtaattctccttggaatttgccctttttgagtttggatcttggttcattctcaa1140gcctcagacagtggttcaaagtttttttcttccatttcaggtgtcgtgaggaagatctct1200agaagctgggtaccttgtgcccgggcgccaccatggagtttgggctgagctggctttttc1260ttgtcgcgattcttaagggtgtccagtgcgacaaaactcacacatgcccaccgtgcccag1320cacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccc1380tcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagacc1440ctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagc1500cgcgggaggagcagtacaacagcacgtaccgtgtggtcagcgtcctcaccgtcctgcacc1560aggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagccc1620ccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccc1680tgcccccatcccgggatgagctgaccaagaaccaggtcagcctgacctgcctggtcaaag1740gcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaact1800acaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctca1860ccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgagg1920ctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaatgagcggccg1980ctcgaggccggcaaggccggatccagacatgataagatacattgatgagtttggacaaac2040cacaactagaatgcagtgaaaaaaatgctttatttgtgaaatttgtgatgctattgcttt2100atttgtaaccattataagctgcaataaacaagttaacaacaacaattgcattcattttat2160gtttcaggttcagggggaggtgtgggaggttttttaaagcaagtaaaacctctacaaatg2220tggtatggctgattatgatccggctgcctcgcgcgtttcggtgatgacggtgaaaacctc2280tgacacatgcagctcccggatacggtcacagcttgtctgtaagcggatgccgggagcaga2340caagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcagccatgaggtcg2400actctagaggatcgatccccgccccggacgaactaaacctgactacgacatctctgcccc2460ttcttcgcggggcagtgcatgtaatcccttcagttggttggtacaacttgccaactgggc2520cctgttccacatgtgacacggggggggaccaaacacaaaggggttctctgactgtagttg2580acatccttataaatggatgtgcacatttgccaacactgagtggctttcatcctggagcag2640actttgcagtctgtggactgcaacacaacattgcctttatgtgtaactcttggctgaagc2700tcttacaccaatgctgggggacatgtacctcccaggggcccaggaagactacgggaggct2760acaccaacgtcaatcagaggggcctgtgtagctaccgataagcggaccctcaagagggca2820ttagcaatagtgtttataaggcccccttgttaaccctaaacgggtagcatatgcttcccg2880ggtagtagtatatactatccagactaaccctaattcaatagcatatgttacccaacggga2940agcatatgctatcgaattagggttagtaaaagggtcctaaggaacagcgatatctcccac3000cccatgagctgtcacggttttatttacatggggtcaggattccacgagggtagtgaacca3060ttttagtcacaagggcagtggctgaagatcaaggagcgggcagtgaactctcctgaatct3120tcgcctgcttcttcattctccttcgtttagctaatagaataactgctgagttgtgaacag3180taaggtgtatgtgaggtgctcgaaaacaaggtttcaggtgacgcccccagaataaaattt3240ggacggggggttcagtggtggcattgtgctatgacaccaatataaccctcacaaacccct3300tgggcaataaatactagtgtaggaatgaaacattctgaatatctttaacaatagaaatcc3360atggggtggggacaagccgtaaagactggatgtccatctcacacgaatttatggctatgg3420gcaacacataatcctagtgcaatatgatactggggttattaagatgtgtcccaggcaggg3480accaagacaggtgaaccatgttgttacactctatttgtaacaaggggaaagagagtggac3540gccgacagcagcggactccactggttgtctctaacacccccgaaaattaaacggggctcc3600acgccaatggggcccataaacaaagacaagtggccactcttttttttgaaattgtggagt3660gggggcacgcgtcagcccccacacgccgccctgcggttttggactgtaaaataagggtgt3720aataacttggctgattgtaaccccgctaaccactgcggtcaaaccacttgcccacaaaac3780cactaatggcaccccggggaatacctgcataagtaggtgggcgggccaagataggggcgc3840gattgctgcgatctggaggacaaattacacacacttgcgcctgagcgccaagcacagggt3900tgttggtcctcatattcacgaggtcgctgagagcacggtgggctaatgttgccatgggta3960gcatatactacccaaatatctggatagcatatgctatcctaatctatatctgggtagcat4020aggctatcctaatctatatctgggtagcatatgctatcctaatctatatctgggtagtat4080atgctatcctaatttatatctgggtagcataggctatcctaatctatatctgggtagcat4140atgctatcctaatctatatctgggtagtatatgctatcctaatctgtatccgggtagcat4200atgctatcctaatagagattagggtagtatatgctatcctaatttatatctgggtagcat4260atactacccaaatatctggatagcatatgctatcctaatctatatctgggtagcatatgc4320tatcctaatctatatctgggtagcataggctatcctaatctatatctgggtagcatatgc4380tatcctaatctatatctgggtagtatatgctatcctaatttatatctgggtagcataggc4440tatcctaatctatatctgggtagcatatgctatcctaatctatatctgggtagtatatgc4500tatcctaatctgtatccgggtagcatatgctatcctcatgcatatacagtcagcatatga4560tacccagtagtagagtgggagtgctatcctttgcatatgccgccacctcccaagggggcg4620tgaattttcgctgcttgtccttttcctgctgcttatcgatgataagctgtcaaacatgag4680aattcttgaagacgaaagggcctcgtgatacgcctatttttataggttaatgtcatgata4740ataatggtttcttagacgtcaggtggcacttttcggggaaatgtgcgcggaacccctatt4800tgtttatttttctaaatacattcaaatatgtatccgctcatgagacaataaccctgataa4860atgcttcaataatattgaaaaaggaagagtatgagtattcaacatttccgtgtcgccctt4920attcccttttttgcggcattttgccttcctgtttttgctcacccagaaacgctggtgaaa4980gtaaaagatgctgaagatcagttgggtgcacgagtgggttacatcgaactggatctcaac5040agcggtaagatccttgagagttttcgccccgaagaacgttttccaatgatgagcactttt5100aaagttctgctatgtggcgcggtattatcccgtgttgacgccgggcaagagcaactcggt5160cgccgcatacactattctcagaatgacttggttgagtactcaccagtcacagaaaagcat5220cttacggatggcatgacagtaagagaattatgcagtgctgccataaccatgagtgataac5280actgcggccaacttacttctgacaacgatcggaggaccgaaggagctaaccgcttttttg5340cacaacatgggggatcatgtaactcgccttgatcgttgggaaccggagctgaatgaagcc5400ataccaaacgacgagcgtgacaccacgatgcctgcagcaatggcaacaacgttgcgcaaa5460ctattaactggcgaactacttactctagcttcccggcaacaattaatagactggatggag5520gcggataaagttgcaggaccacttctgcgctcggcccttccggctggctggtttattgct5580gataaatctggagccggtgagcgtgggtctcgcggtatcattgcagcactggggccagat5640ggtaagccctcccgtatcgtagttatctacacgacggggagtcaggcaactatggatgaa5700cgaaatagacagatcgctgagataggtgcctcactgattaagcattggtaactgtcagac5760caagtttactcatatatactttagattgatttaaaacttcatttttaatttaaaaggatc5820taggtgaagatcctttttgataatctcatgaccaaaatcccttaacgtgagttttcgttc5880cactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctg5940cgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccg6000gatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagatacca6060aatactgttcttctagtgtagccgtagttaggccaccacttcaagaactctgtagcaccg6120cctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcg6180tgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctga6240acggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatac6300ctacagcgtgagctatgagaaagcgccacgcttcccgaagggagaaaggcggacaggtat6360ccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcc6420tggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtga6480tgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttc6540ctggccttttgctggccttttgctcacatgaagctgtccctgatggtcgtcatctacctg6600cctggacagcatggcctgcaacgcgggcatcccgatgccgccggaagcgagaagaatcat6660aatggggaaggccatccagcctcgcgtcgcgaacgccagcaagacgtagcccagcgcgtc6720ggccccgagatgcgccgcgtgcggctgctggagatggcggacgcgatggatatgttctgc6780caagggttggtttgcgcattcacagttctccgcaagaattgattggctccaattcttgga6840gtggtgaatccgttagcgaggtgccgccctgcttcatccccgtggcccgttgctcgcgtt6900tgctggcggtgtcactggccccgtgggttagggacggggtcccccatggggaatggttta6960tggttcgtgggggttattattttgggcgttgcgtggggtcaggtccacgactggactgag7020cagacagacccatggtttttggatggcctgggcatggaccgcatgtactggcgcgacacg7080aacaccgggcgtctgtggctgccaaacacccccgacccccaaaaaccaccgcgcggattt7140ctggcgtgccaagctagtcgaccaattctcatgtttgacagcttatcatcgcagatccgg7200gcaacgttgttgccattgctgcaggcgcagaactggtaggtatggaagatct7252<210>134<211>19<212>prt<213>人工序列(artificialsequence)<400>134alathrasnpheserleuleulysglnalaglyaspvalglugluasn151015proglypro<210>135<211>19<212>prt<213>人工序列(artificialsequence)<400>135alathrasnpheserleuleulysglnalaglyaspvalglugluasn151015proglypro当前第1页12