CN111138533A - 针对甲型肝炎病毒的单域抗体及其衍生蛋白 - Google Patents

针对甲型肝炎病毒的单域抗体及其衍生蛋白 Download PDF

Info

Publication number
CN111138533A
CN111138533A CN201911398827.1A CN201911398827A CN111138533A CN 111138533 A CN111138533 A CN 111138533A CN 201911398827 A CN201911398827 A CN 201911398827A CN 111138533 A CN111138533 A CN 111138533A
Authority
CN
China
Prior art keywords
seq
gly
ser
ala
tyr
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201911398827.1A
Other languages
English (en)
Other versions
CN111138533B (zh
Inventor
苏志鹏
姚尧
王乐飞
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nanjing Rongjiekang Biotechnology Co ltd
Original Assignee
Nanjing Rongjiekang Biotechnology Co ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Nanjing Rongjiekang Biotechnology Co ltd filed Critical Nanjing Rongjiekang Biotechnology Co ltd
Priority to CN201911398827.1A priority Critical patent/CN111138533B/zh
Publication of CN111138533A publication Critical patent/CN111138533A/zh
Application granted granted Critical
Publication of CN111138533B publication Critical patent/CN111138533B/zh
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
    • C07K16/10—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from RNA viruses
    • C07K16/1009—Picornaviridae, e.g. hepatitis A virus
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56983—Viruses
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/565—Complementarity determining region [CDR]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/569—Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody®
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/30—Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
    • G01N2333/08—RNA viruses
    • G01N2333/085—Picornaviridae, e.g. coxsackie virus, echovirus, enterovirus
    • G01N2333/10—Hepatitis A virus
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Virology (AREA)
  • Immunology (AREA)
  • Molecular Biology (AREA)
  • Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Medicinal Chemistry (AREA)
  • Urology & Nephrology (AREA)
  • Biomedical Technology (AREA)
  • Organic Chemistry (AREA)
  • Biophysics (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Genetics & Genomics (AREA)
  • Microbiology (AREA)
  • Communicable Diseases (AREA)
  • Food Science & Technology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Peptides Or Proteins (AREA)

Abstract

本发明涉及生物技术或免疫学技术领域，涉及针对甲型肝炎病毒的单域抗体及其衍生蛋白。所述的单域抗体的CDR1的氨基酸序列如SEQ ID NO:83‑SEQ ID NO:117中的任意一条所示；所述的CDR2的氨基酸序列如SEQ ID NO:118‑SEQ ID NO:143中的任意一条所示；所述的CDR3的氨基酸序列如SEQ ID NO:144‑SEQ ID NO:174中的任意一条所示。本发明的有益效果是：传统通过检测甲肝病毒抗体（HAV‑IgM,HAV‑IgG）用于甲肝病毒抗原（HAV‑Ag）的间接检测的方法，其检测步骤多，影响因素也多。本发明提供了一种基于ELISA不需要通过RT‑PCR的直接检测甲肝病毒抗原（HAV‑Ag）的检测方法。

Description

针对甲型肝炎病毒的单域抗体及其衍生蛋白
技术领域
本发明涉及生物技术或免疫学技术领域，涉及针对甲型肝炎病毒的单域抗体及其衍生蛋白。
背景技术
抗体(Antibody,Ab)即免疫球蛋白(Immunoglobulin,Ig)是血液和组织液中的一类糖蛋白，由B细胞接收抗原刺激后增殖分化生成的浆细胞产生，主要存在与血清等体液中，能与相应抗原特异性地结合，是介导体液免疫的重要效应分子。抗体除了因其作为重要效应分子介导特异性体液免疫应答外，抗体在疾病的防治特别是感染性疾病的防治方面扮演者重要角色，Behring创立血清疗法因此获得医学与生理学诺贝尔奖。此后，以多克隆、单克隆及基因工程抗体技术人工制备的抗体逐步应用于临床。仅2011年抗药药物就以480亿美元的销售额领跑全球药品市场，占整个生物制药市场的34.4％。至2017年，美国FDA和欧盟EMA批准的治疗性抗体药物有76个，主要用于肿瘤、自身免疫病等疾病的治疗。
抗体作为可以特异性识别某一个蛋白(抗原)的分子，除了在疾病预防和治疗方面的作用强大之外，其作为检测工具在检验、科学研究方面的用途也不言而喻。酶联免疫吸附实验(ELISA)正是利用抗体/抗原结合以实现检测目的一个重要方法。目前常用的ELISA检测方法有直接法、间接法、竞争法等，其基本原理均为抗原抗体结合反应。以此方法为基础ELISA试剂盒在检验检疫等场合的用途也十分广泛。部分检验检疫方法对于抗体的亲和力和稳定性要求非常苛刻，这就要求所使用的抗体在与其特异性结合的抗原亲和力要非常高，稳定性要非常好，并且这些抗体的使用和储存环境要更为广泛才更有利于检测方法的推广。传统单抗在抗原亲和力方面表现良好，并且虽然现在可以通过添加保护剂或改变缓冲液的方法提升其稳定性，但是其储存条件要求依然相对苛刻，一般应在低温运输或保存。而单域抗体在亲和力和稳定性方面就表现的更为优秀，单域抗体在未进行亲和力成熟之前，其与抗原的亲和力可达到pM级别，并且其室温稳定性良好，可以降低运输和储存成本。此外，单域抗体可以在原核表达系统中可以大量可溶性表达，降低生产成本，并且提高抗体批件稳定性。
甲型病毒性肝炎简称甲肝，是一种由甲型肝炎病毒(HAV)感染引起的以肝脏炎症病变为主的传染病。1974年Feinstone等通过免疫电镜技术在甲肝患者粪便中发现甲肝病毒(HAV)。甲肝病毒是直径为27～32nm的小RNA病毒，其结构含有VP1～VP4四种多肽，基因组为单股正链RNA，约7500个核苷酸。 HAV的传播途径为粪－口途径，主要在发展中国家流行。我国是甲肝的高流行国家。提高人群的免疫水平是预防甲肝的主要措施，随着甲肝灭活疫苗应用覆盖面不断扩大，甲肝的暴发流行得到有效的控制。因其临床表现与其他肝炎相似，因此甲肝的实验室诊断结果就显得尤为重要。在甲肝急性期患者血清中可以查出抗HAV-IgM及IgG，患者患病后2周抗HAV-IgM阳性率为100％，持续2～3周，然后迅速下降。因此，检测抗HAV-IgM可作为急性期的诊断指标。抗HAV-IgG是甲肝的主要中和抗体，一般可从病人血清中查出，此种抗体存在于患者体内很长时间甚至终身。甲肝的实验室诊断方法主要有RT-PCR 法、RT-PCR-ELISA法、间接ELISA法、SPA协同凝集试验等。RT-PCR法灵敏度高，特异性强。但需专门的仪器设备，操作过程复杂，且对RNA的提取过程要求较严格；RT-PCR-ELISA法先通过RT-PCR扩增甲肝病毒RNA，再用ELISA法检测扩增产物。在保证了特异性的同时，也提高了检测的灵敏度。但是试验必须建立在RT-PCR的基础上，检测成本高；SPA协同凝集试验利用金黄色葡萄球菌细胞壁中的A蛋白(SPA)能特异性的与HAV-IgG的Fc段结合，当待检样本中含有HAV-Ag时，即出现反向间接凝集反应。该试验灵敏度和特异性较低，结果判断人为影响因素大，仅可作为设备受限时的初步检测方法；一般的ELISA法利用固相载体有吸附免疫活性蛋白的特性，在已包被抗人μ链的酶标板中，加入HAV抗体待检血清，若待检血清为甲肝感染急性期，则血清中的IgM抗体被其抗体人μ链抗体捕获，再加入酶标抗体，底物显色后，判断待检血清者为甲肝感染患者。但该方法只能检测已感染甲肝病毒的患者血清的IgM抗体，不能直接定量或半定量患者体内甲肝病毒；所以直接用甲肝病毒抗体检测待检样品中甲肝病毒含量可以更直接、准确的反映患者血清内或者其他样品中的甲肝病毒含量，但目前市面上直接检测甲肝病毒的定量或半定量ELISA试剂盒产品很少，并且已有的直接检测试剂盒的捕获抗体选择的是兔源多克隆抗体，由于兔多抗的制备批间差异很大，导致抗体的亲和力差异很大，这就造成了试剂盒检测的生产批间稳定性很差，重复性不好。所以选择生产工艺稳定并且亲和力更高的甲肝病毒抗体是准确检测样本甲肝病毒含量的重点。
发明内容
为了克服上述缺陷，本发明的目的是提供特异性针对甲型肝炎病毒的单域抗体并建立基于抗甲型肝炎病毒单域抗体及其衍生蛋白，首先，在获得高特异性的单域抗体后通过原核表达或者酵母表达便可以获得，产量和纯化效率高并且批间质量稳定，其次，单域抗体的亲和力比传统抗体有一定优势，可以提升抗原捕获能力和检测效果，提高检测的灵敏度，此外，单域抗体属于单克隆抗体，这就能保证检测试剂的单一性和可控性，更好的控制检测试剂盒的检测标准。
单域抗体作为单克隆抗体的一类，其比传统单抗的亲和力普遍更高，稳定性更好，并且它也具备传统单抗的优点。因此，本发明就是利用上述优势，获得针对甲型肝炎病毒的抗体，这些获得的抗体既能够在普通的ELISA实验中作为检测工具对各类样品中的甲型肝炎病毒进行定性分析，也可以被开发成ELISA检测试剂盒，对人群中的甲型肝炎病毒进行定量分析，用于甲肝的临床诊断。
为解决上述技术问题，本发明采用如下技术方案：针对甲型肝炎病毒的单域抗体,其特征在于，所述的单域抗体的序列包括互补决定区CDR；所述互补决定区CDR包括CDR1、CDR2和CDR3的氨基酸序列；所述的CDR1的氨基酸序列如SEQ ID NO:83-SEQ ID NO:117中的任意一条所示；所述的CDR2的氨基酸序列如SEQ ID NO:118-SEQ ID NO:143中的任意一条所示；所述的CDR3的氨基酸序列如SEQ ID NO:144-SEQ ID NO:174中的任意一条所示。
上述所有序列，可以替换为与该序列具有“至少80％同源性”的序列或仅一个或少数几个氨基酸替换的序列；优选为“至少85％同源性”，更优选为“至少90％同源性”，更优选为“至少95％同源性”，最优选为“至少98％同源性”。
针对甲型肝炎病毒的单域抗体,所述的单域抗体的序列包括互补决定区CDR；所述互补决定区CDR 包括CDR1、CDR2和CDR3的氨基酸序列；所述的单域抗体的互补决定区CDR的序列为下述(1)-(39) 中的一种：
(1)SEQ ID NO:83所示的CDR1，SEQ ID NO:118所示的CDR2，SEQ ID NO:144所示的CDR3(对应于抗体1A2、1B4)；
(2)SEQ ID NO:84所示的CDR1，SEQ ID NO:119所示的CDR2，SEQ ID NO:145所示的CDR3(对应于抗体1A4)；
(3)SEQ ID NO:84所示的CDR1，SEQ ID NO:119所示的CDR2，SEQ ID NO:156所示的CDR3(对应于抗体4A1)；
(4)SEQ ID NO:84所示的CDR1，SEQ ID NO:119所示的CDR2，SEQ ID NO:169所示的CDR3(对应于抗体4A9)；
(5)SEQ ID NO:85所示的CDR1，SEQ ID NO:120所示的CDR2，SEQ ID NO:146所示的CDR3(对应于抗体1A6)；
(6)SEQ ID NO:86所示的CDR1，SEQ ID NO:121所示的CDR2，SEQ ID NO:147所示的CDR3(对应于抗体1A7)；
(7)SEQ ID NO:87所示的CDR1，SEQ ID NO:122所示的CDR2，SEQ ID NO:148所示的CDR3(对应于抗体1B1)；
(8)SEQ ID NO:88所示的CDR1，SEQ ID NO:123所示的CDR2，SEQ ID NO:149所示的CDR3(对应于抗体1B10)；
(9)SEQ ID NO:89所示的CDR1，SEQ ID NO:124所示的CDR2，SEQ ID NO:150所示的CDR3(对应于抗体1C4)；
(10)SEQ ID NO:90所示的CDR1，SEQ ID NO:124所示的CDR2，SEQ ID NO:150所示的CDR3(对应于抗体1C9)；
(11)SEQ ID NO:91所示的CDR1，SEQ ID NO:125所示的CDR2，SEQ ID NO:151所示的CDR3(对应于抗体1D6)；
(12)SEQ ID NO:92所示的CDR1，SEQ ID NO:126所示的CDR2，SEQ ID NO:152所示的CDR3(对应于抗体1D7)；
(13)SEQ ID NO:92所示的CDR1，SEQ ID NO:126所示的CDR2，SEQ ID NO:153所示的CDR3(对应于抗体1E5)；
(14)SEQ ID NO:92所示的CDR1，SEQ ID NO:126所示的CDR2，SEQ ID NO:173所示的CDR3(对应于抗体4F11)；
(15)SEQ ID NO:93所示的CDR1，SEQ ID NO:126所示的CDR2，SEQ ID NO:152所示的CDR3(对应于抗体1F4；
(16)SEQ ID NO:94所示的CDR1，SEQ ID NO:127所示的CDR2，SEQ ID NO:154所示的CDR3(对应于抗体1G4)；
(17)SEQ ID NO:95所示的CDR1，SEQ ID NO:128所示的CDR2，SEQ ID NO:155所示的CDR3(对应于抗体1G11)；
(18)SEQ ID NO:96所示的CDR1，SEQ ID NO:129所示的CDR2，SEQ ID NO:156所示的CDR3(对应于抗体1H1)；
(19)SEQ ID NO:97所示的CDR1，SEQ ID NO:126所示的CDR2，SEQ ID NO:152所示的CDR3(对应于抗体1H2)；
(20)SEQ ID NO:98所示的CDR1，SEQ ID NO:130所示的CDR2，SEQ ID NO:157所示的CDR3(对应于抗体1H3)；
(21)SEQ ID NO:99所示的CDR1，SEQ ID NO:126所示的CDR2，SEQ ID NO:158所示的CDR3(对应于抗体1H9)；
(22)SEQ ID NO:100所示的CDR1，SEQ ID NO:131所示的CDR2，SEQ ID NO:159所示的CDR3(对应于抗体2A10)；
(23)SEQ ID NO:101所示的CDR1，SEQ ID NO:132所示的CDR2，SEQ ID NO:160所示的CDR3(对应于抗体2B2、4E5)；
(24)SEQ ID NO:102所示的CDR1，SEQ ID NO:126所示的CDR2，SEQ ID NO:152所示的CDR3(对应于抗体2B8；
(25)SEQ ID NO:103所示的CDR1，SEQ ID NO:133所示的CDR2，SEQ ID NO:161所示的CDR3(对应于抗体2B12)；
(26)SEQ ID NO:104所示的CDR1，SEQ ID NO:134所示的CDR2，SEQ ID NO:162所示的CDR3(对应于抗体2C3)；
(27)SEQ ID NO:105所示的CDR1，SEQ ID NO:135所示的CDR2，SEQ ID NO:163所示的CDR3(对应于抗体2E2)；
(28)SEQ ID NO:106所示的CDR1，SEQ ID NO:136所示的CDR2，SEQ ID NO:164所示的CDR3(对应于抗体2G1)；
(29)SEQ ID NO:107所示的CDR1，SEQ ID NO:137所示的CDR2，SEQ ID NO:165所示的CDR3(对应于抗体3B10)；
(30)SEQ ID NO:108所示的CDR1，SEQ ID NO:138所示的CDR2，SEQ ID NO:166所示的CDR3(对应于抗体3D7)；
(31)SEQ ID NO:109所示的CDR1，SEQ ID NO:139所示的CDR2，SEQ ID NO:167所示的CDR3(对应于抗体3D11)；
(32)SEQ ID NO:110所示的CDR1，SEQ ID NO:140所示的CDR2，SEQ ID NO:168所示的CDR3(对应于抗体4A3)；
(33)SEQ ID NO:111所示的CDR1，SEQ ID NO:118所示的CDR2，SEQ ID NO:144所示的CDR3(对应于抗体4A11)；
(34)SEQ ID NO:112所示的CDR1，SEQ ID NO:126所示的CDR2，SEQ ID NO:152所示的CDR3(对应于抗体4D1)；
(35)SEQ ID NO:113所示的CDR1，SEQ ID NO:141所示的CDR2，SEQ ID NO:170所示的CDR3(对应于抗体4D10)；
(36)SEQ ID NO:114所示的CDR1，SEQ ID NO:142所示的CDR2，SEQ ID NO:171所示的CDR3(对应于抗体4E7)；
(37)SEQ ID NO:115所示的CDR1，SEQ ID NO:126所示的CDR2，SEQ ID NO:172所示的CDR3(对应于抗体4F5)；
(38)SEQ ID NO:116所示的CDR1，SEQ ID NO:143所示的CDR2，SEQ ID NO:174所示的CDR3(对应于抗体4G5)；
(39)SEQ ID NO:117所示的CDR1，SEQ ID NO:118所示的CDR2，SEQ ID NO:144所示的CDR3(对应于抗体4H4)。
优选的，针对甲型肝炎病毒的单域抗体，所述单域抗体为SEQ ID NO:1-41中的任意一条所示的抗体链。
单域抗体链的编码序列分别如SEQ ID NO：42-82所示。
所述的抗体序列还包括框架区FR；所述框架区FR包括FR1、FR2、FR3和FR4的氨基酸序列；框架区FR的氨基酸序列分别为：SEQ ID NO:175-195任意一条所示的FR1；SEQ ID NO:196-222任意一条所示的FR2；SEQ ID NO:223-252任意一条所示的FR3；SEQ ID NO:253-254任意一条所示的FR4。上述所有FR1、FR2、FR3和FR4的序列可以替换为其保守序列变体。
本发明的又一个目的是提供一种核苷酸分子，该核苷酸分子编码所述的针对甲型肝炎病毒的单域抗体，其核苷酸序列如SEQ ID NO：42-82中的任意一条所示。
本发明还提供一种表达载体，其包含所述核苷酸分子。
本发明还涉及一种宿主细胞，其可以表达出所述的针对甲型肝炎病毒的单域抗体，或其包含所述的表达载体。
本发明还提供针对甲型肝炎病毒的单域抗体的Fc融合抗体。
本发明还公开特异性针对甲型肝炎病毒的单域抗体在HAV-ELISA检测试剂盒的制备中的应用，由前述针对甲型肝炎病毒的单域抗体在检测人群中甲型肝炎病毒含量的半定量和定量检测方法。
相对于现有技术，本发明的有益效果是：单域抗体作为甲型肝炎病毒检测抗体，其与传统单克隆抗体相比，其亲和力更高，与现有检测试剂盒中多克隆抗体相比，其成分更加单一，批间稳定性更好，质量控制更容易实施。从而使实际的甲型肝炎病毒含量检测结果可信度更高。
附图说明
图1是对所构建的单域抗体噬菌体展示文库进行目的片段插入率检测的PCR产物电泳图，从左至右，泳道1和泳道17是DNA分子marker，泳道2-16、泳道18-33是从构建的针对甲型肝炎病毒单域抗体文库中随机挑取的不同克隆的PCR产物；
图2是对所构建的单域抗体文库进行生物淘选的具体结果，其中P/N＝生物淘选中阳性孔洗脱下的噬菌体感染TG1细菌后生长的单克隆细菌数/阳性孔洗脱下的噬菌体感染TG1细菌后生长的单克隆细菌数，该参数在富集发生后会逐渐增大；I/E＝生物淘选中每轮加入阳性孔的噬菌体总量/生物淘选中每轮从阳性孔洗脱出的噬菌体总量，该参数在富集发生后会逐渐趋近于1；
图3是部分表达纯化后的针对甲型肝炎病毒的单域抗体，蛋白粗液经镍柱树脂亲和层析纯化后的 SDS-PAGE电泳图，从左至右，泳道1是蛋白marker，泳道2-14是经镍离子耦联的琼脂糖树脂一步亲和层析纯化后的单域抗体，其中VHH1-13与SEQ ID NO.1-13一一对应；
图4是纯化后的纳米抗体与甲肝病毒结合的ELISA检测结果；
图5是纯化后的纳米抗体对甲肝病毒衣壳蛋白的配对ELISA检测结果；
图6是融合表达载体的示意图。
具体实施方式
下面结合实施例对本发明做进一步的详细说明，以令本领域技术人员参照说明书文字能够据以实施。
针对甲型肝炎病毒的纳米抗体
单域抗体(sdAb，也被研发者Ablynx称为纳米抗体或VHH)是本领域技术人员公知的。单域抗体为其互补决定区是单域多肽的一部分的抗体。因此，单域抗体包含单个互补决定区(单个CDR1、单个CDR2 和单个CDR3)。单域抗体的实例为仅有重链的抗体(该抗体天然不包含轻链)、衍生自常规抗体的单域抗体和工程化抗体。
单域抗体可以衍生自任何物种，包括小鼠、人、骆驼、美洲驼、山羊、兔和牛。例如，天然存在的VHH分子可以衍生自骆驼科物种(例如骆驼、单峰骆驼、美洲驼和原驼)提供的抗体。像完整的抗体一样，单域抗体能够选择性地结合特定抗原。单域抗体可以仅含有免疫球蛋白链的可变结构域，该结构域具有CDR1、CDR2和CDR3以及框架区。纳米抗体的分子量仅为约12-15kDa，比由两条重链和两条轻链组成的普通抗体的分子量(150-160kDa)小得多。
值得一提的是，本发明中，与本发明公开的CDR1-3的序列同源性高的序列，也可以得到针对甲肝病毒的纳米抗体。在一些实施例中，与(1)-(39)中的序列具有“至少80％同源性”的序列，或者“至少85％同源性”、“至少90％同源性”、“至少95％同源性”、“至少98％同源性”的序列都可以实现发明目的(即衍生蛋白)。
在一些实施例中，与(1)-(39)中的序列相比仅仅替换一个或少数几个氨基酸的序列，例如，包含1、2、3、4、5、6、7、8、9或10个保守氨基酸取代，也可以实现发明目的。实际上，在确定两个氨基酸序列之间的序列同一性程度或在确定单域抗体中的CDR1、CDR2和CDR3组合时，技术人员可以考虑所谓的“保守”氨基酸取代，在取代情况下，所述取代将优选为保守氨基酸取代，所述保守氨基酸，其通常可以被描述为氨基酸残基被具有相似化学结构的另一个氨基酸残基替代的氨基酸取代，并且该取代对多肽的功能、活性或其它生物学性质几乎没有或基本上没有影响。所述保守氨基酸取代在本领域是通用的，例如保守氨基酸取代是以下(a)-(d)组内的一个或少数几个氨基酸被同一组内另一个或少数几个氨基酸所取代：(a)极性带负电残基及其不带电酰胺：Asp、Asn、Glu、Gln；(b) 极性带正电残基：His、Arg、Lys；(c)芳香族残基：Phe、Trp、Tyr；(d)脂肪族非极性或弱极性残基：Ala、Ser、Thr、Gly、Pro、Met、Leu、Ile、Val、Cys。特别优选的保守氨基酸取代如下：Asp 被Glu取代；Asn被Gln或His取代；Glu被Asp取代；Gln被Asn取代；His被Asn或Gln取代；Arg 被Lys取代；Lys被Arg、Gln取代；Phe被Met、Leu、Tyr取代；Trp被Tyr取代；Tyr被Phe、Trp 取代；Ala被Gly或Ser取代；Ser被Thr取代；Thr被Ser取代；Gly被Ala或Pro取代；Met被Leu、 Tyr或Ile取代；Leu被Ile或Val取代；Ile被Leu或Val取代；Val被Ile或Leu取代；Cys被Ser 取代。另外，本领域技术人员知晓，单域抗体的创造性体现在CDR1-3区，而框架区序列FR1-4并非是不可改变的，FR1-4的序列可以采取本发明公开的序列的保守序列变体。
本发明采用甲型肝炎病毒灭活疫苗来免疫阿拉善双峰驼，经过7次免疫之后提取该双峰驼外周血淋巴细胞并建立甲型肝炎病毒的单域抗体文库。在抗体筛选过程中，将甲型肝炎病毒衣壳蛋白偶联生物素(VP2-VP3-VP1-Biotin)，将中性亲和素蛋白偶联在酶标板上，利用生物素和中性亲和素结合的特性，使甲型肝炎病毒衣壳蛋白间接展示在酶标板表面，使其抗原表位得以暴露，然后利用噬菌体展示技术筛选甲型肝炎病毒灭活疫苗免疫后的单域抗体基因库(骆驼重链抗体噬菌体展示基因库)，获得了能在大肠杆菌中高效表达的单域抗体株。
将筛选获得的单域抗体在大肠杆菌中表达后通过镍离子偶联的琼脂糖进行纯化，将纯化后的纳米抗体进行分析。
下面结合具体实施例，进一步阐述本发明。
实施例1：针对甲型肝炎病毒的单域抗体文库的构建：
(1)使用1ml甲型肝炎病毒灭活疫苗免疫一只内蒙古阿拉善双峰驼，每周一次，共连续免疫7次，免疫过程中刺激B细胞表达特异性的纳米抗体；(2)免疫结束后，提取骆驼外周血淋巴细胞100ml并提取总RNA；(3)合成cDNA并利用套式PCR扩增VHH；(4)利用限制性内切酶Pst I及Not I酶切20μg pMECS噬菌体展示载体及10μg VHH并连接两种片段；(5)将连接产物转化至电转感受态细胞TG1中，构建针对甲肝病毒衣壳蛋白噬菌体展示文库并测定库容，文库库容的大小约为2×109；同时，通过菌落 PCR检测所建文库中目的片段的正确插入率，图1显示菌落PCR结果，随机的选取30个克隆做菌落 PCR，结果显示插入率达到93.3％。
实施例2：甲型肝炎病毒衣壳蛋白的表达：
(1)从NCBI的蛋白数据库中查到甲型肝炎病毒衣壳蛋白的序列信息(AccesionNumber:P08617.1)； (2)经过Uniprot分析，选取VP2-VP3-VP1作为目的表达片段；(3)在VP2-VP3-VP1的5’端添加kozak 增强序列(5’-GGATCGAACCCTT-3’)及IgG kappa的信号肽(METDTLLLWVLLLWVPGSTGD)，3’端添加6*His标签融合表达；(4)使用实施例8中的所述方案进行真核表达；(5)参照实施例9所述方案使用镍柱亲和层析进行纯化；(6)参照实施例9方法进行SDS-PAGE分析，确定蛋白纯度在95％以上，且浓度在0.5mg/mL以上后，低温保存备用。
实施例3：针对甲型肝炎病毒单域抗体的筛选：
(1)取200μL重组TG1细胞至2×TY培养基中培养，期间加入40μL辅助噬菌体VCSM13侵染TG1 细胞，并培养过夜以扩增噬菌体，次日利用PEG/NaCl沉淀噬菌体，离心收集扩增噬菌体；(2)将稀释在100mM pH 8.3的NaHCO3中的中性亲和素蛋白500ug偶联在酶标板上，4℃放置过夜，同时设立阴性对照孔；(3)第二天加入100μL生物素标记的甲型肝炎病毒衣壳蛋白(VP2-VP3-VP1-Biotin)，室温孵育2小时，阴性对照孔加入100μL PBS；(4)2小时后，加入100μL的3％的脱脂乳，室温封闭2h； (5)封闭结束后，加入100μl扩增后噬菌体文库(大约2×1011个噬菌体颗粒)，室温作用1h；(6)作用1 小时后，用PBS+0.05％Tween-20洗5遍，以洗掉未结合的噬菌体；(7)用终浓度为25mg/mL的胰蛋白酶将与甲型肝炎病毒衣壳蛋白特异性结合的噬菌体解离下，并感染处于对数生长期的大肠杆菌TG1细胞，37℃培养1h，产生并收集噬菌体用于下一轮的筛选，相同筛选过程重复3轮，逐步得到富集，富集效果如图2所示，P/N值从第一轮的2.2到第三轮约为30，I/E质从第一轮的13333到第三轮约为 333，证明文库针对甲型肝炎病毒衣壳蛋白具有非常明显的富集。
实施例4：用噬菌体的酶联免疫方法(ELISA)筛选特异性阳性克隆:
(1)根据上述单域抗体筛选方法对甲型肝炎病毒纳米抗体进行3轮筛选，筛选结束后，针对甲型肝炎病毒衣壳蛋白的噬菌体富集因子达到10以上(约为30)，从筛选获得的阳性克隆中挑选400个单菌落分别接种于含100μg/mL氨苄青霉素的TB培养基的96深孔板中，并设置空白对照，37℃培养至对数期后，加入终浓度为1mM的IPTG，28℃培养过夜；(2)利用渗透涨破法获得粗提抗体；将中性亲和素蛋白稀释至100mM pH 8.3的NaHCO3中并将100μg中性亲和素蛋白在酶标板中4℃包被过夜，次日向酶标板内加入100ug VP2-VP3-VP1-Biotin蛋白；(3)将上述步骤中获得的抗体粗提液取100uL转移至加入抗原的ELISA板上，室温孵育1h；(4)用PBST洗去未结合的抗体，加入100ul经1:2000稀释后的 Mouse anti-HAtagantibody(鼠抗HA抗体，Thermo Fisher)，在室温孵育1h；(5)用PBST洗去未结合的抗体，加入100ul经1:20000稀释后的Anti-Rabbit HRP conjugate(山羊抗兔辣根过氧化物酶标记抗体，购自于Thermo Fisher)，在室温孵育1h；(6)用PBST洗去未结合的抗体，加入辣根过氧化物酶显色液，37℃下反应15min后，加入中止液，于酶标仪上450nm波长处，读取吸收值；(7)当样品孔OD 值大于对照孔5倍以上时，判定为阳性克隆孔；(8)将阳性克隆孔的菌转摇在含有100μg/ul氨苄青霉素的LB培养基中以便提取质粒并进行测序。
根据序列比对软件Vector NTI分析各个克隆株的基因序列，把CDR1，CDR2，CDR3序列相同的株视为同一克隆株，而序列不同的株视为不同克隆株，最终获得特异性针对甲型肝炎病毒的单域抗体。其抗体的氨基酸序列为FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4结构，构成整个VHH。获得的单域抗体重组质粒可以在原核系统中进行表达，最终获得单域抗体蛋白。
其中，其中SEQ ID NO.1-41依次为抗体编号1A2、1A4、1A6、1A7、1B1、1B4、1B10、1C4、1C9、1D6、1D7、1E5、1F4、1G4、1G11、1H1、1H2、1H3、1H9、2A10、2B2、2B8、2B12、2C3、2E2、2G1、 3B10、3D7、3D11、4A1、4A3、4A9、4A11、4D1、4D10、4E5、4E7、4F5、4F11、4G5、4H4。
41种单域抗体序列的CDR1-3的序列如表1-2所示，41种单域抗体的FR1-3序列如表3-4所示。其中41种单域抗体中，除了抗体1B1的FR4序列为SEQ ID NO:254(GRGTQVTVSS)以外，其它40 种抗体的FR4序列均为SEQ ID NO:253(GQGTQVTVSS)。
表1 41种单域抗体的CDR1-2序列
Figure BDA0002347006380000091
Figure BDA0002347006380000101
表2 41种单域抗体的CDR3序列
Figure BDA0002347006380000102
Figure BDA0002347006380000111
表3 41种单域抗体的FR1、FR2序列
Figure BDA0002347006380000112
Figure BDA0002347006380000121
表4 41种单域抗体的FR3序列
Figure BDA0002347006380000122
Figure BDA0002347006380000131
实施例5：甲型肝炎病毒的特异性单域抗体在宿主菌大肠杆菌中的纯化及表达
(1)将上述测序分析所获得不同克隆株的质粒(pMECS-VHH)电转化到大肠杆菌HB2151中，并将其涂布在LB+amp+glucose即含有氨苄青霉素和葡萄糖的培养平板上，37℃培养过夜；(2)挑选单个菌落接种在5mL含有岸边青霉素的LB培养液中，37℃摇床培养过夜；(3)接种1mL的过夜培养菌种至330mL TB培养液中，37℃摇床培养，培养到OD600nm值达到0.6-0.9时，加入1M IPTG，28℃摇床培养过夜；(4)离心，收集大肠杆菌，利用渗透胀破法，获得抗体粗提液；(5)通过镍柱亲和层析法纯化出抗体，纯化后的单域抗体，如图3所示。
实施例6：甲型肝炎病毒的特异性单域抗体的Fc融合抗体真核表达载体的构建
将实施例4中获得的目标序列亚克隆至真核表达载体中：(1)将实施例4中筛选出来的抗体经 Sanger测序得到其核苷酸序列；(2)通过序列合成的方式将密码子优化后的上述核苷酸序列(SEQ ID NO.42-82)合成至本公司设计改造的载体RJK-V4-hFC中(即SEQ IDNO.42-82分别插入载体RJK-V4-hFC 的多克隆位点MCS，RJK-V4-hFc载体的构建过程如实施例11所述)；(3)将构建好的重组真核表达载体转化至DH5α大肠杆菌中，培养进行质粒大提，去除内毒素；(4)将大提后的质粒再进行序列测序鉴定；(5)将确定无误后的重组载体准备后续真核细胞转染表达。
实施例7：甲型肝炎病毒的特异性单域抗体的Fc融合抗体在悬浮ExpiCHO-S细胞中表达
(1)转染前3天以2.5×105/ml细胞传代和扩大培养ExpiCHO-STM细胞，计算出的所需的细胞体积转移至装有新鲜的已预热的120ml(终体积)的ExpiCHOTM表达培养基的500ml摇瓶中；使细胞浓度达到约4×10 6-6×106活细胞/mL；(2)在转染前一天，将ExpiCHO-STM细胞稀释浓度至3.5×10 6活细胞 /mL，使细胞过夜培养；(3)转染当天，测定细胞密度和活细胞百分比。转染之前细胞密度应达到约7 ×106-10×106活细胞/mL；(4)用预热至37℃新鲜的ExpiCHOTM表达培养基将细胞稀释至6×106个活细胞/mL。计算出的所需的细胞体积转移至装有新鲜的已预热的100ml(终体积)的ExpiCHOTM表达培养基的500ml摇瓶中；(5)使轻轻颠倒混匀ExpiFectamineTMCHO试剂，用3.7ml OptiPROTM培养基稀释 ExpiFectamineTMCHO试剂，回荡或混匀；(6)用冷藏的4ml OptiPROTM培养基稀释质粒DNA，回荡混匀； (7)将ExpiFectamine CHO/质粒DNA复合物室温孵育1-5分钟，然后轻轻加入制备的细胞悬液中，加入过程中轻轻回荡摇瓶，所述的质粒DNA为实施例6制得的甲型肝炎病毒的特异性单域抗体的Fc 融合抗体真核表达载体；(8)将细胞在37℃、8％CO2、加湿的空气中震荡培养；(9)转染后第1天 (18-22小时后)添加600ul ExpiFectamineTMCHO Enhancer和24ml ExpiCHOfeed。(10)在转染后约8 天(细胞活率低于70％)收集上清。
实施例8：甲型肝炎病毒的特异性单域抗体的Fc融合抗体在悬浮293F细胞中的表达
重组单域抗体表达实验流程(以500ml摇瓶为例)：
1、转染前3天以2.5×105/ml细胞传代和扩大培养293F细胞，计算出的所需的细胞体积转移至装有新鲜的已预热的120ml(终体积)的OPM-293CD05 Medium培养基的500ml摇瓶中。使细胞浓度达到约2×106-3×106活细胞/mL。
2、转染当天，测定细胞密度和活细胞百分比。转染之前细胞密度应达到约2×106-3×106活细胞/ mL。
3、用预热的OPM-293CD05 Medium将细胞稀释至1×106个活细胞/mL。计算出所需的细胞体积转移至装有新鲜的已预热的100ml(终体积)的培养基的500ml摇瓶中。
4、用4ml Opti-MEM培养基稀释PEI(1mg/ml)试剂，回荡或吹打混匀；用4ml Opt-MEM培养基稀释质粒DNA，回荡混匀，并用0.22um的滤头过滤。室温孵育5min；所述的质粒DNA为实施例6 制得的甲型肝炎病毒的特异性单域抗体的Fc融合抗体真核表达载体。
5、将稀释的PEI试剂加入稀释的DNA中，颠倒混匀。将PEI/质粒DNA复合物室温孵育15-20 分钟，然后轻轻加入制备的细胞悬液中，加入过程中轻轻回荡摇瓶。
6、将细胞在37℃、5％CO2、120rpm震荡培养。
7、转染后第24h、72h添加5ml OPM-CHO PFF05补料。
8、在转染后约7天(细胞活率低于70％)收集上清。
实施例9：人源Fc重组单域抗体的纯化
1、将实施例7或8中获得蛋白表达上清用0.45μm的一次性滤头过滤除掉不可溶杂质；
2、将上述滤液使用蛋白纯化仪进行亲和层析纯化，利用人源Fc与Protein A结合的能力，使用偶联Protein A的琼脂糖填料进行纯化；
3、将滤液通过1mL/分钟的流速流穿Protein A预装柱，该步骤中滤液中的目标蛋白会与填料结合；
4、通过低盐和高盐缓冲液将柱上结合的杂质蛋白洗涤；
5、用低pH缓冲液将柱上结合的目标蛋白进行系统；
6、将洗脱液迅速加入pH9.0的Tris-HCl溶液，进行中和；
7、将上述中和后的蛋白溶液透析后，进行SDS-PAGE分析，确定蛋白纯度在95％以上，且浓度在0.5mg/mL以上后，低温保存备用。
实施例10：特异性针对甲型肝炎病毒衣壳蛋白的重组单域抗体与甲型肝炎病毒亲和力的ELISA检测以及检测标准曲线的绘制
(1)将实施例9中纯化获得重组抗体(实施例7制得的特异性单域抗体的Fc融合抗体经实施例9纯化而得)对甲型肝炎病毒的亲和力进行初步检测；
(2)首先使用定量的标准品测定重组单域抗体与甲肝病毒的亲和力步骤如下(即下面的步骤 (3)-(9)；
(3)将100ng/100μL甲型肝炎病毒衣壳蛋白样品包被到ELISA板材上；
(4)使用脱脂奶粉对上述包被的板材进行封闭；
(5)加入实施例9中获得的针对甲型肝炎病毒的重组单域抗体；
(6)加入特异性针对人源Fc的检测抗体(HRP标记)
(7)加入显色底物TMB；
(8)加入终止液终止反应；
(9)测量OD450值(结果如图4所示，可见针对甲型肝炎病毒的单域抗体均与甲型肝炎病毒具有较强的亲和力)；
(10)然后挑选ELISA结合力最好的重组抗体进行甲型肝炎病毒含量标准曲线的绘制。
(11)将0-100ng/100μL的甲型肝炎病毒样品包被到ELISA板材上；
(12)使用脱脂奶粉对步骤(11)的包被板材进行封闭；
(13)加入步骤(10)中获得的效果最佳的针对甲型肝炎病毒的重组单域抗体；
(14)加入特异性针对人源Fc的检测抗体(HRP标记)
(15)加入显色底物TMB；
(16)加入终止液终止反应；
(17)测量OD450值，根据结果绘制甲型肝炎病毒含量和OD450关系曲线图，选择曲线图线性范围区段进行线性回归计算，作为标准曲线进行其他样品中甲型肝炎病毒含量的测定。
实施例10：甲型肝炎病毒的特异性单域抗体在HAV-ELISA试剂盒中的应用(双抗夹心法)
(1)将实施例9和实施例5中获得单域抗体通过双抗夹心法检测目标样品中甲型肝炎病毒的含量；
(2)首先使用标准品测定并绘制甲型肝炎病毒含量与OD450的标准曲线，步骤如下；
(3)将实施例9中鉴定、纯化得到单域抗体作为捕获抗体包被到ELISA板材上；
(4)使用脱脂奶粉对上述包被的板材进行封闭；
(5)加入标准样品孵育；
(6)加入实施例5中获得的单域抗体作为检测抗体(带有HA标签)，孵育；
(7)加入抗HA的抗体孵育(HRP标记)；
(8)加入显色底物TMB；
(9)加入终止液终止反应；
(10)测量OD450值，并作出比值分析，结果如图5所示；
(11)然后基于上述步骤获得的标准曲线对待检样品进行检测。
实施例11：纳米抗体真核表达载体RJK-V4-hFc的构建
所提及纳米抗体通用的目标载体RJK-V4-hFC，为本公司在invitrogen商业化载体pcDNA3.4(载体资料链接
https://assets.thermofisher.com/TFS-Assets/LSG/manuals/pcdna3_4_topo_ta_cloning_kit_man. pdf)的基础上融合了人源IgG的重链编码序列(NCBI AccessionNo.:AB776838.1)中的Fc区段后改造而来的，即该载体包含了IgG重链的铰链区(Hinge)CH2和CH3区。具体改造方案如下：
(1)选取pcDNA3.4上的限制性酶切位点XbaI和AgeI；
(2)在Fc片段编码序列的5’端和3’端通过重叠PCR的方式分别引入多克隆位点(MCS， Multiple Cloning Site)和6×His标签，如图6所示；
(3)使用分别带有XbaI和AgeI酶切位点的一对引物通过PCR的方式将上述片段扩增；
(4)使用限制性内切酶XbaI和AgeI分别酶切pcDNA3.4和(3)中的重组DNA片段；
(5)将酶切后的载体和插入片段在T4连接酶的作用下连接，然后将连接产物转化至大肠杆菌，扩增，测序核实，获得重组质粒。
尽管本发明的实施方案已公开如上，但其并不仅仅限于说明书和实施方式中所列运用，它完全可以被适用于各种适合本发明的领域，对于熟悉本领域的人员而言，可容易地实现另外的修改，因此在不背离权利要求及等同范围所限定的一般概念下，本发明并不限于特定的细节。
序列表
<110> 南京融捷康生物科技有限公司
<120> 针对甲型肝炎病毒的单域抗体及其衍生蛋白
<130> GY-03-2019-006
<141> 2019-12-26
<160> 256
<170> SIPOSequenceListing 1.0
<210> 1
<211> 127
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 1
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Tyr Ser Gly Arg Ser Asp
20 25 30
Tyr Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ala Val Ile Phe Thr Gly Gly Gly Arg Thr Trp Tyr Ala Asn Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Gln Gly Asn Ala Glu Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Leu Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Ala Asp Tyr Val Pro Val Gly Arg Arg Tyr Ala Val Leu Asp Pro
100 105 110
Asp Tyr Thr Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 2
<211> 121
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 2
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ala Ala Tyr Ser Asp Tyr
20 25 30
Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Arg Val
35 40 45
Ala Thr Phe Leu Arg Asp Gly Ser Ala Asp Tyr Ala Ala Ser Val Thr
50 55 60
Gly Arg Phe Thr Ile Ser Lys Asp Asn Val Lys Asn Thr Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys Ala
85 90 95
Ala Asp Tyr Trp Gly Tyr Gly Tyr Asn Cys Arg Phe Glu Tyr Trp Gly
100 105 110
Gln Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 3
<211> 127
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 3
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Asp Tyr Ser Thr Tyr
20 25 30
Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ala Ser Ile Ser Thr Val Gly Thr Thr Ser Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Asn Met Tyr Tyr Cys Ala
85 90 95
Ser Leu Asp Arg Arg Met Tyr Gly Gly Ile Trp Tyr Cys Asp Leu Arg
100 105 110
Arg Tyr Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 4
<211> 115
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 4
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Pro Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Asn Ser Gly Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Leu Asn Gly Leu Lys Thr Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Leu Asp Ser Arg Gly Thr Gln Arg Gly Gln Gly Thr Gln Val Thr
100 105 110
Val Ser Ser
115
<210> 5
<211> 121
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 5
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Tyr Ser Gly Ser Gly Asn Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ala Asp Trp Thr Gly Arg Thr Leu His Glu Tyr Asn Tyr Leu Gly
100 105 110
Arg Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 6
<211> 127
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 6
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Tyr Ser Gly Arg Ser Asp
20 25 30
Tyr Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ala Val Ile Phe Thr Gly Gly Gly Arg Thr Trp Tyr Ala Asn Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Leu Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Ala Asp Tyr Val Pro Val Gly Arg Arg Tyr Ala Val Leu Asp Pro
100 105 110
Asp Tyr Thr Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 7
<211> 123
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 7
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Tyr Ile Ser His Pro Tyr
20 25 30
Cys Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ala Gly Ile Ser Ile Gly Gly Gly Gly Thr Phe Thr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val His
65 70 75 80
Leu Arg Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Ala Arg Gly Gly Tyr Tyr Cys Thr Ala Ser Ala Ser Tyr Asn Tyr
100 105 110
Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 8
<211> 126
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 8
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Tyr Ser Arg Tyr
20 25 30
Ser Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ala Ser Leu Asp Ser Asp Gly Ala Thr Ile Tyr Thr Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ala Lys Asp Asn Ala Lys Asn Thr Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Met Tyr Tyr Cys Ala
85 90 95
Ala Gln Arg Ile Arg Trp Val Ala Phe Val Ser Leu Arg Ser Ala Asp
100 105 110
Phe Asp Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 9
<211> 126
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 9
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Ala Tyr Ser Arg Tyr
20 25 30
Ser Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ala Ser Leu Asp Ser Asp Gly Ala Thr Ile Tyr Thr Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ala Lys Asp Asn Ala Lys Asn Thr Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Met Tyr Tyr Cys Ala
85 90 95
Ala Gln Arg Ile Arg Trp Val Ala Phe Val Ser Leu Arg Ser Ala Asp
100 105 110
Phe Asp Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 10
<211> 129
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 10
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Trp Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Ser Lys Arg Ala
20 25 30
Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Trp Val
35 40 45
Ala Thr Ile Ser Arg Pro Gly Gly Ala Thr Tyr Ser Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Glu Asn Met Ala Phe
65 70 75 80
Leu Ile Leu Asn Ser Leu Lys Ala Glu Asp Ala Ala Ile Tyr Tyr Cys
85 90 95
Ala Phe Arg Val Ile Asp Pro Phe Asp Thr Cys Ser Ser Arg Arg Arg
100 105 110
Asp Gly Asp Phe Pro Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser
115 120 125
Ser
<210> 11
<211> 125
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 11
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Ala Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Asn Ser Tyr Ser Arg Asn
20 25 30
Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Gln Gly Val
35 40 45
Ala Ala Ile Ser Pro Gly Gly Ser Phe Lys Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Gln Pro Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Ala Ala Pro Phe Ala Cys Asp Gly Asp Trp Tyr Ser Gly Arg Trp
100 105 110
Asn Asn Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 12
<211> 125
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 12
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Ala Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Asn Ser Tyr Ser Arg Asn
20 25 30
Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Gln Gly Val
35 40 45
Ala Ala Ile Ser Pro Gly Gly Ser Phe Lys Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Gln Pro Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Ala Ala Pro Phe Ala Cys Asp Gly Asp Trp His Ser Gly Arg Trp
100 105 110
Asn Asn Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 13
<211> 125
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 13
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr His Ser Ser Tyr
20 25 30
Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Gln Gly Val
35 40 45
Ala Ala Ile Ser Pro Gly Gly Ser Phe Lys Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Gln Pro Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Ala Ala Pro Phe Ala Cys Asp Gly Asp Trp Tyr Ser Gly Arg Trp
100 105 110
Asn Asn Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 14
<211> 130
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 14
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Tyr Ser Ser Asn
20 25 30
Cys Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ala Val Ile Tyr Thr Gly Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Ala Asp Leu Arg Ala Ala Pro Tyr Gly Gly Ile Gly Tyr Phe Pro
100 105 110
Ala Ala Thr Asp Phe Gly Tyr Trp Gly Gln Gly Thr Gln Val Thr Val
115 120 125
Ser Ser
130
<210> 15
<211> 131
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 15
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ala Cys Ala Ala Ser Gly Tyr Ala Tyr Ser Arg Tyr
20 25 30
Tyr Met Gly Trp Phe Arg Gln Val Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ala Thr Ile Ser Thr Gly Gly Gly Thr Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Val Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Thr Pro Glu Asp Thr Ala Ile Tyr Tyr Cys
85 90 95
Ala Ala Arg Val Gly Ser Pro Phe Gly Gly Ala Pro Asp Leu Ser Ser
100 105 110
Leu Arg Pro Glu Arg Tyr Ser Tyr Trp Gly Gln Gly Thr Gln Val Thr
115 120 125
Val Ser Ser
130
<210> 16
<211> 118
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 16
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Gly Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Val Val Ser Gly Tyr Thr Ile Ser Ser Leu
20 25 30
Cys Met Gly Trp Phe Arg Arg Thr Glu Arg Glu Ala Val Ala Arg Ile
35 40 45
Asp Arg Asp Gly Gly Thr Arg Tyr Ala Asp Pro Val Lys Gly Arg Phe
50 55 60
Thr Ile Ser Lys Asp Gly Ala Lys Asn Thr Leu Tyr Leu Gln Met Asn
65 70 75 80
Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys Ala Ala Asp Tyr
85 90 95
Trp Gly Tyr Gln Tyr Asn Cys Arg Phe Glu Tyr Trp Gly Gln Gly Thr
100 105 110
Gln Val Thr Val Ser Ser
115
<210> 17
<211> 125
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 17
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Asp Thr Asn Arg Gln Arg
20 25 30
Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Glu Gly Arg Glu Gly Val
35 40 45
Ala Ala Ile Ser Pro Gly Gly Ser Phe Lys Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Gln Pro Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Ala Ala Pro Phe Ala Cys Asp Gly Asp Trp Tyr Ser Gly Arg Trp
100 105 110
Asn Asn Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 18
<211> 127
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 18
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Thr Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Val Asn Thr Lys
20 25 30
Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ala Thr Ile Tyr Thr Gly Thr Ser Thr Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Arg Asn Thr Ile Tyr
65 70 75 80
Leu Gln Met Asp Ser Val Lys Pro Asp Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Ala Thr Thr Tyr Ile Gly Ser Trp Cys Gly Leu Arg Ser Pro Leu
100 105 110
Asp Tyr Asn Asn Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 19
<211> 125
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 19
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Ala Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Tyr Ser Thr Asn
20 25 30
Cys Met Gly Trp Leu Arg Gln Ala Pro Gly Lys Glu Arg Gln Gly Val
35 40 45
Ala Ala Ile Ser Pro Gly Gly Ser Phe Lys Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Ala Ala Pro Phe Ala Cys Asp Gly Asp Trp His Ser Arg Arg Trp
100 105 110
Asn Asn Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 20
<211> 126
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 20
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Tyr Thr Gly Asp Thr Ser
20 25 30
Ala Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ala Ser Ile Tyr Arg Val Ser Ser Ala Thr Phe Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Gln Asp Lys Ala Asn Asp Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Asn Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ala Arg Ser Thr Ala Val Thr Leu Arg Gln Leu Thr Gln Gln Ala
100 105 110
Tyr Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 21
<211> 126
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 21
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Pro Gly Tyr Thr Tyr Ser Ser Ala
20 25 30
Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ala Thr Ile Gly Ser Asp Gly Ser Thr Ser Tyr Thr Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Lys Asp Asn Val His Asn Asn Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys Ala
85 90 95
Ala Thr Ile Thr Thr Val Thr Met Cys Arg Arg Arg Asp Gly Arg Leu
100 105 110
Phe Gly His Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 22
<211> 125
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 22
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ser Val Ser Gly Tyr Thr Tyr Ser Tyr Asn
20 25 30
Cys Val Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Gln Gly Val
35 40 45
Ala Ala Ile Ser Pro Gly Gly Ser Phe Lys Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Gln Pro Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Ala Ala Pro Phe Ala Cys Asp Gly Asp Trp Tyr Ser Gly Arg Trp
100 105 110
Asn Asn Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 23
<211> 119
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 23
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ala Gly Phe Arg Phe Ser Arg Val
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Arg Ile Asn Asp Asp Gly Ser Trp Arg Asp Ala Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Thr Gly Phe Pro Ser Gly Gly Asn Trp Ile Gln Arg Gly Gln Gly
100 105 110
Thr Gln Val Thr Val Ser Ser
115
<210> 24
<211> 128
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 24
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Asp Thr Ala Asn Ser Asn
20 25 30
Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Glu Glu Arg Glu Trp Val
35 40 45
Ala Ser Ile Val Ser Ala Ser Gly Phe Ile Tyr Tyr Ala Asp Ala Val
50 55 60
Arg Gly Arg Phe Thr Val Ser Gln Asp Lys Ser Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Ala Lys Pro Arg Pro Arg Phe Leu Thr Val Pro Ala Gly Cys Gly
100 105 110
Tyr Glu Tyr Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 25
<211> 125
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 25
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Tyr Thr Ile Ala
20 25 30
Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Ile Ala Ala Ile
35 40 45
Tyr Thr Leu Ala Gly Gly Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg
50 55 60
Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Ser Leu Gln Met
65 70 75 80
Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys Ala Ala Ala
85 90 95
Ile Gly Ala Gly Tyr Gly Arg Asn Trp Leu Leu Ser Glu Arg Trp Tyr
100 105 110
Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 26
<211> 127
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 26
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Tyr Ser Ser Ser
20 25 30
Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ala Thr Ile Asp Ser Phe Gly Asn Thr Lys Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Lys Asp Asn Ala Lys Asn Thr Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys Thr
85 90 95
Ala Asp Met Asp Gly Arg Trp Gly Gly Tyr Cys Val Arg Ser Ala Glu
100 105 110
Asp Phe Ala Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 27
<211> 121
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 27
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Thr Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly His Gly Ser Cys Thr Tyr
20 25 30
Asp Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys Ser Arg Glu Phe Val
35 40 45
Ser Val Ile Met Ser Asp Gly Thr Thr Ala Tyr Thr Ser Ser Val Lys
50 55 60
Asp Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Ser Thr Val Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Arg Tyr Tyr Cys Glu
85 90 95
Ala Val Leu Arg Gly Arg Cys Thr Arg Ser Gly Phe Thr Tyr Trp Gly
100 105 110
Gln Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 28
<211> 126
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 28
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Ser Thr Tyr Ser Arg Arg
20 25 30
Tyr Ile Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ala Thr Ile Ser Thr Gly Gly Gly Phe Arg Ser Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Ala Gly Ser Asn Val Asp Arg Arg Trp Ser Leu Leu Ser Arg Asp
100 105 110
Tyr Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 29
<211> 120
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 29
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Val Val Ser Gly Tyr Thr Ile Asp Arg Leu
20 25 30
Cys Thr Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val
35 40 45
Ser Ser Thr Ser Thr Asp Gly Ile Thr Arg Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Val Ser Ser Asp Lys Ala Lys Asp Thr Val Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Phe Cys His
85 90 95
Phe Gly Gly Arg Trp Leu Thr Pro Arg Ala Cys Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 30
<211> 121
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 30
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ala Ala Tyr Ser Asp Tyr
20 25 30
Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Arg Val
35 40 45
Ala Thr Phe Leu Arg Asp Gly Ser Ala Asp Tyr Ala Ala Ser Val Thr
50 55 60
Gly Arg Phe Thr Ile Ser Lys Asp Asn Val Lys Asn Thr Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys Ala
85 90 95
Ala Asp Tyr Trp Gly Tyr Gln Tyr Asn Cys Arg Phe Glu Tyr Trp Gly
100 105 110
Gln Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 31
<211> 132
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 31
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Tyr Thr Val Ser Arg Ser
20 25 30
Cys Met Ala Trp Phe Arg Gln Val Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ala Ser Ile Ser Ser Asp Gly Gly Ile Thr Thr Tyr Val Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Val Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Thr Gly Val Leu Gly Phe Phe Ser Asn Leu Cys Arg Ser Thr Val
100 105 110
Val Pro Pro Thr Ala Arg Phe Arg Tyr Trp Gly Gln Gly Thr Gln Val
115 120 125
Thr Val Ser Ser
130
<210> 32
<211> 121
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 32
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ala Ala Tyr Ser Asp Tyr
20 25 30
Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Arg Val
35 40 45
Ala Thr Phe Leu Arg Asp Gly Ser Ala Asp Tyr Ala Ala Ser Val Thr
50 55 60
Gly Arg Phe Thr Ile Ser Lys Asp Asn Val Lys Asn Thr Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys Ala
85 90 95
Val Asp Tyr Trp Gly Tyr Gly Tyr Asn Cys Arg Phe Glu Tyr Trp Gly
100 105 110
Gln Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 33
<211> 127
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 33
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Thr Cys Ala Val Ser Gly Tyr Ser Tyr Ser Ser Asn
20 25 30
His Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ala Val Ile Phe Thr Gly Gly Gly Arg Thr Trp Tyr Ala Asn Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser His Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Leu Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Ala Asp Tyr Val Pro Val Gly Arg Arg Tyr Ala Val Leu Asp Pro
100 105 110
Asp Tyr Thr Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 34
<211> 125
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 34
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Trp Ser Ser Thr
20 25 30
Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Gln Gly Val
35 40 45
Ala Ala Ile Ser Pro Gly Gly Ser Phe Lys Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Gln Pro Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Ala Ala Pro Phe Ala Cys Asp Gly Asp Trp Tyr Ser Gly Arg Trp
100 105 110
Asn Asn Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 35
<211> 126
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 35
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Pro Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr
20 25 30
Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Val Leu Glu Trp Val
35 40 45
Ser Phe Ile Asn Ser Gly Gly Gly Gly Thr Ala Tyr Ala Asp Pro Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Leu Asn Ser Leu Lys Thr Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Lys Gly Leu Gln Tyr Asp Thr Tyr Cys Ser Ser Gln Tyr Cys Tyr
100 105 110
Ala Asn Pro Gly Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 36
<211> 126
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 36
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Thr Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Tyr Ser Ser Ala
20 25 30
Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ala Thr Ile Gly Ser Asp Gly Ser Thr Ser Tyr Thr Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Lys Asp Asn Val His Asn Asn Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys Ala
85 90 95
Ala Thr Ile Thr Thr Val Thr Met Cys Arg Arg Arg Asp Gly Arg Leu
100 105 110
Phe Gly His Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 37
<211> 126
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 37
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Pro Tyr Ser Thr Tyr
20 25 30
Tyr Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ala Ala Val Tyr Pro Gly Val Gly Leu Lys Arg Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Gln Asp Asp Ala Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Leu Tyr Tyr Cys
85 90 95
Ala Val Asp Gln Ala Val Ser Leu Gly Trp Glu Leu Ser Pro Gly Arg
100 105 110
Tyr Arg Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 38
<211> 125
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 38
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Thr Gly Gly
1 5 10 15
Ser Leu Lys Leu Ser Cys Val His Ser Gly Tyr Ser Tyr Ser Ser Val
20 25 30
Cys Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ala Ala Ile Ser Pro Gly Gly Ser Phe Lys Tyr Tyr Val Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Gly Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Thr Ser Arg Ser Gly Cys Arg Asp Asp Phe Phe Gln Tyr Glu Tyr
100 105 110
Asp Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 39
<211> 125
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 39
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Ala Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Asn Ser Tyr Ser Arg Asn
20 25 30
Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Gln Gly Val
35 40 45
Ala Ala Ile Ser Pro Gly Gly Ser Phe Lys Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Gln Pro Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Ala Ala Pro Leu Ala Cys Asp Gly Asp Trp Tyr Ser Gly Arg Trp
100 105 110
Asn Asn Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 40
<211> 128
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 40
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Pro Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Tyr Ser Gly Ser
20 25 30
Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Glu Val
35 40 45
Ala Thr Ile Tyr Thr Asp Asn Gly Phe Lys Tyr Tyr Thr Glu Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Gln Asp Lys Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ala Asp Gly Gly Thr Gly Leu Trp Tyr Thr Cys Ala Arg Gly Gln
100 105 110
Ile Ser His Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 41
<211> 127
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 41
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Thr Val Ser Gly Tyr Ser Tyr Ser Ser Asp
20 25 30
Tyr Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val
35 40 45
Ala Val Ile Phe Thr Gly Gly Gly Arg Thr Trp Tyr Ala Asn Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Leu Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Ala Asp Tyr Val Pro Val Gly Arg Arg Tyr Ala Val Leu Asp Pro
100 105 110
Asp Tyr Thr Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120 125
<210> 42
<211> 381
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 42
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
agctgcgccg tgagcggcta cagcggcagg agcgactaca tgggctggtt caggcaggcc 120
cccggcaagg agagggaggg cgtggccgtg atcttcaccg gcggcggcag gacctggtac 180
gccaacagcg tgaagggcag gttcaccatc agccagggca acgccgagaa caccgtgtac 240
ctgcagatga acagcctgaa gctggaggac accgccatgt actactgcgc cgccgactac 300
gtgcccgtgg gcaggaggta cgccgtgctg gaccccgact acacctactg gggccagggc 360
acccaggtga ccgtgagcag c 381
<210> 43
<211> 363
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 43
caggtgcagc tggtggagag cggcggcggc ctggtgcagc ccggcggcag cctgaggctg 60
agctgcgccg ccagcggcgc cgcctacagc gactactgca tgggctggtt caggcaggcc 120
cccggcaagg agagggagag ggtggccacc ttcctgaggg acggcagcgc cgactacgcc 180
gccagcgtga ccggcaggtt caccatcagc aaggacaacg tgaagaacac cctgtacctg 240
cagatgaaca gcctgaagcc cgaggacacc gccatgtact actgcgccgc cgactactgg 300
ggctacggct acaactgcag gttcgagtac tggggccagg gcacccaggt gaccgtgagc 360
agc 363
<210> 44
<211> 381
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 44
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
agctgcgccg ccagcggcta cgactacagc acctactgca tgggctggtt caggcaggcc 120
cccggcaagg agagggaggg cgtggccagc atcagcaccg tgggcaccac cagctacgcc 180
gacagcgtga agggcaggtt caccatcagc agggacaacg ccaagaacac cgtgtacctg 240
cagatgaaca gcctgaagac cgaggacacc aacatgtact actgcgccag cctggacagg 300
aggatgtacg gcggcatctg gtactgcgac ctgaggaggt acaactactg gggccagggc 360
acccaggtga ccgtgagcag c 381
<210> 45
<211> 345
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 45
caggtgcagc tggtggagag cggcggcggc agcgtgcagc ccggcggcag cctgaggctg 60
agctgcgccg ccagcggctt caccttcagc agctacccca tgaactgggt gaggcaggcc 120
cccggcaagg gcctggagtg ggtgagcgcc atcaacagcg gcggcggcag cacctactac 180
gccgacagcg tgaagggcag gttcaccatc agcagggaca acgccaagaa caccctgtac 240
ctgcagctga acggcctgaa gaccgaggac accgccatgt actactgcgc cctggacagc 300
aggggcaccc agaggggcca gggcacccag gtgaccgtga gcagc 345
<210> 46
<211> 363
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 46
caggtgcagc tggtggagag cggcggcggc ctggtgcagc ccggcggcag cctgaggctg 60
agctgcgccg ccagcggctt caccttcagc aggtacgaca tgagctgggt gaggcagccc 120
cccggcaagg gcctggagtg ggtgagcgcc atctacagcg gcagcggcaa cacctactac 180
gccgacagcg tgaagggcag gttcaccatc agcagggaca acgccaagaa caccctgtac 240
ctgcagatga acagcctgaa gaccgaggac accgccgtgt actactgcgc cgccgactgg 300
accggcagga ccctgcacga gtacaactac ctgggcaggg gcacccaggt gaccgtgagc 360
agc 363
<210> 47
<211> 381
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 47
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
agctgcgccg tgagcggcta cagcggcagg agcgactaca tgggctggtt caggcaggcc 120
cccggcaagg agagggaggg cgtggccgtg atcttcaccg gcggcggcag gacctggtac 180
gccaacagcg tgaagggcag gttcaccatc agccaggaca acgccaagaa caccgtgtac 240
ctgcagatga acagcctgaa gctggaggac accgccatgt actactgcgc cgccgactac 300
gtgcccgtgg gcaggaggta cgccgtgctg gaccccgact acacctactg gggccagggc 360
acccaggtga ccgtgagcag c 381
<210> 48
<211> 369
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 48
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
agctgcgccg tgagcggcta catcagccac ccctactgca tggcctggtt caggcaggcc 120
cccggcaagg agagggaggg cgtggccggc atcagcatcg gcggcggcgg caccttcacc 180
gccgacagcg tgaagggcag gttcaccatc agccaggaca acgccaagaa caccgtgcac 240
ctgaggatga acagcctgaa gcccgaggac accgccatgt actactgcgc cgccaggggc 300
ggctactact gcaccgccag cgccagctac aactactggg gccagggcac ccaggtgacc 360
gtgagcagc 369
<210> 49
<211> 378
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 49
caggtgcagc tggtggagag cggcggcggc agcgtgcagc ccggcggcag cctgaggctg 60
agctgcgccg ccagcggcta cacctacagc aggtacagca tgggctggtt caggcaggcc 120
cccggcaagg agagggaggg cgtggccagc ctggacagcg acggcgccac catctacacc 180
gacagcgtga agggcaggtt caccatcgcc aaggacaacg ccaagaacac cctgtacctg 240
cagatgaaca gcctgaagcc cgaggacacc ggcatgtact actgcgccgc ccagaggatc 300
aggtgggtgg ccttcgtgag cctgaggagc gccgacttcg actactgggg ccagggcacc 360
caggtgaccg tgagcagc 378
<210> 50
<211> 378
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 50
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
agctgcgccg ccagcggcta cgcctacagc aggtacagca tgggctggtt caggcaggcc 120
cccggcaagg agagggaggg cgtggccagc ctggacagcg acggcgccac catctacacc 180
gacagcgtga agggcaggtt caccatcgcc aaggacaacg ccaagaacac cctgtacctg 240
cagatgaaca gcctgaagcc cgaggacacc ggcatgtact actgcgccgc ccagaggatc 300
aggtgggtgg ccttcgtgag cctgaggagc gccgacttcg actactgggg ccagggcacc 360
caggtgaccg tgagcagc 378
<210> 51
<211> 387
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 51
caggtgcagc tggtggagag cggcggcggc tgggtgcagg ccggcggcag cctgaggctg 60
agctgcgccg ccagcggcgt gaccagcaag agggcctgca tgggctggtt caggcaggcc 120
cccggcaagg agagggagtg ggtggccacc atcagcaggc ccggcggcgc cacctacagc 180
gccgacagcg tgaagggcag gttcaccatc agccaggaca acgccgagaa catggccttc 240
ctgatcctga acagcctgaa ggccgaggac gccgccatct actactgcgc cttcagggtg 300
atcgacccct tcgacacctg cagcagcagg aggagggacg gcgacttccc ctactggggc 360
cagggcaccc aggtgaccgt gagcagc 387
<210> 52
<211> 375
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 52
caggtgcagc tggtggagag cggcggcggc agcgcccagg ccggcggcag cctgaggctg 60
agctgcgccg ccagcggcaa cagctacagc aggaactgca tgggctggtt caggcaggcc 120
cccggcaagg agaggcaggg cgtggccgcc atcagccccg gcggcagctt caagtactac 180
gccgacagcg tgaagggcag gttcaccatc agcagggaca acgccaagaa caccgtgtac 240
ctgcagatga acagcctgca gcccgaggac accgccatgt actactgcgc cgccgccccc 300
ttcgcctgcg acggcgactg gtacagcggc aggtggaaca actggggcca gggcacccag 360
gtgaccgtga gcagc 375
<210> 53
<211> 375
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 53
caggtgcagc tggtggagag cggcggcggc agcgcccagg ccggcggcag cctgaggctg 60
agctgcgccg ccagcggcaa cagctacagc aggaactgca tgggctggtt caggcaggcc 120
cccggcaagg agaggcaggg cgtggccgcc atcagccccg gcggcagctt caagtactac 180
gccgacagcg tgaagggcag gttcaccatc agcagggaca acgccaagaa caccgtgtac 240
ctgcagatga acagcctgca gcccgaggac accgccatgt actactgcgc cgccgccccc 300
ttcgcctgcg acggcgactg gcacagcggc aggtggaaca actggggcca gggcacccag 360
gtgaccgtga gcagc 375
<210> 54
<211> 375
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 54
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
agctgcgccg ccagcggcta cacccacagc agctactgca tgggctggtt caggcaggcc 120
cccggcaagg agaggcaggg cgtggccgcc atcagccccg gcggcagctt caagtactac 180
gccgacagcg tgaagggcag gttcaccatc agcagggaca acgccaagaa caccgtgtac 240
ctgcagatga acagcctgca gcccgaggac accgccatgt actactgcgc cgccgccccc 300
ttcgcctgcg acggcgactg gtacagcggc aggtggaaca actggggcca gggcacccag 360
gtgaccgtga gcagc 375
<210> 55
<211> 390
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 55
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
agctgcgccg ccagcggcta cacctacagc agcaactgca tggcctggtt caggcaggcc 120
cccggcaagg agagggaggg cgtggccgtg atctacaccg gcggcggcag cacctactac 180
gccgacagcg tgaagggcag gttcaccatc agccaggaca acgccaagaa caccgtgtac 240
ctgcagatga acagcctgaa gcccgaggac accgccatgt actactgcgc cgccgacctg 300
agggccgccc cctacggcgg catcggctac ttccccgccg ccaccgactt cggctactgg 360
ggccagggca cccaggtgac cgtgagcagc 390
<210> 56
<211> 393
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 56
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
gcctgcgccg ccagcggcta cgcctacagc aggtactaca tgggctggtt caggcaggtg 120
cccggcaagg agagggaggg cgtggccacc atcagcaccg gcggcggcac cacctactac 180
gccgacagcg tgaagggcag gttcaccgtg agcagggaca acgccaagaa caccctgtac 240
ctgcagatga acagcctgac ccccgaggac accgccatct actactgcgc cgccagggtg 300
ggcagcccct tcggcggcgc ccccgacctg agcagcctga ggcccgagag gtacagctac 360
tggggccagg gcacccaggt gaccgtgagc agc 393
<210> 57
<211> 354
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 57
caggtgcagc tggtggagag cggcggcggc agcggccaac ccggcggcag cctgaggctg 60
agctgcgtgg tgagcggcta caccatcagc agcctgtgca tgggctggtt caggaggacc 120
gagagggagg ccgtggccag gatcgacagg gacggcggca ccaggtacgc cgaccccgtg 180
aagggcaggt tcaccatcag caaggacggc gccaagaaca ccctgtacct gcagatgaac 240
agcctgaagc ccgaggacac cgccatgtac tactgcgccg ccgactactg gggctaccag 300
tacaactgca ggttcgagta ctggggccag ggcacccagg tgaccgtgag cagc 354
<210> 58
<211> 375
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 58
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
agctgcgccg ccagcggcga caccaacagg cagaggtgca tgggctggtt caggcaggcc 120
cccggcgagg gcagggaggg cgtggccgcc atcagccccg gcggcagctt caagtactac 180
gccgacagcg tgaagggcag gttcaccatc agcagggaca acgccaagaa caccgtgtac 240
ctgcagatga acagcctgca gcccgaggac accgccatgt actactgcgc cgccgccccc 300
ttcgcctgcg acggcgactg gtacagcggc aggtggaaca actggggcca gggcacccag 360
gtgaccgtga gcagc 375
<210> 59
<211> 381
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 59
caggtgcagc tggtggagag cggcggcggc agcgtgcaga ccggcggcag cctgaggctg 60
agctgcgccg ccagcggcta caccgtgaac accaagtgca tgggctggtt caggcaggcc 120
cccggcaagg agagggaggg cgtggccacc atctacaccg gcaccagcac cacctactac 180
gccgacagcg tgaagggcag gttcaccatc agcagggaca acgccaggaa caccatctac 240
ctgcagatgg acagcgtgaa gcccgacgac accgccatgt actactgcgc cgccaccacc 300
tacatcggca gctggtgcgg cctgaggagc cccctggact acaacaactg gggccagggc 360
acccaggtga ccgtgagcag c 381
<210> 60
<211> 375
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 60
caggtgcagc tggtggagag cggcggcggc agcgcccagg ccggcggcag cctgaggctg 60
agctgcgccg ccagcggcta cacctacagc accaactgca tgggctggct gaggcaggcc 120
cccggcaagg agaggcaggg cgtggccgcc atcagccccg gcggcagctt caagtactac 180
gccgacagcg tgaagggcag gttcaccatc agcagggaca acgccaagaa caccgtgtac 240
ctgcagatga acagcctgaa gcccgaggac accgccatgt actactgcgc cgccgccccc 300
ttcgcctgcg acggcgactg gcacagcagg aggtggaaca actggggcca gggcacccag 360
gtgaccgtga gcagc 375
<210> 61
<211> 378
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 61
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
agctgcgtgg ccagcggcta caccggcgac accagcgcca tggcctggtt caggcaggcc 120
cccggcaagg agagggaggg cgtggccagc atctacaggg tgagcagcgc caccttctac 180
gccgacagcg tgaagggcag gttcaccatc agccaggaca aggccaacga cagcctgtac 240
ctgcagatga acaacctgaa gcccgaggac accgccgtgt actactgcgc cgccaggagc 300
accgccgtga ccctgaggca gctgacccag caggcctaca actactgggg ccagggcacc 360
caggtgaccg tgagcagc 378
<210> 62
<211> 378
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 62
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
agctgcgccg cccccggcta cacctacagc agcgcctgca tgggctggtt caggcaggcc 120
cccggcaagg agagggaggg cgtggccacc atcggcagcg acggcagcac cagctacacc 180
gacagcgtga agggcaggtt caccatcagc aaggacaacg tgcacaacaa cctgtacctg 240
cagatgaaca gcctgaagcc cgaggacacc gccatgtact actgcgccgc caccatcacc 300
accgtgacca tgtgcaggag gagggacggc aggctgttcg gccactgggg ccagggcacc 360
caggtgaccg tgagcagc 378
<210> 63
<211> 375
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 63
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
agctgcagcg tgagcggcta cacctacagc tacaactgcg tgggctggtt caggcaggcc 120
cccggcaagg agaggcaggg cgtggccgcc atcagccccg gcggcagctt caagtactac 180
gccgacagcg tgaagggcag gttcaccatc agcagggaca acgccaagaa caccgtgtac 240
ctgcagatga acagcctgca gcccgaggac accgccatgt actactgcgc cgccgccccc 300
ttcgcctgcg acggcgactg gtacagcggc aggtggaaca actggggcca gggcacccag 360
gtgaccgtga gcagc 375
<210> 64
<211> 357
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 64
caggtgcagc tggtggagag cggcggcggc ctggtgcagc ccggcggcag cctgaggctg 60
agctgcgccg ccgccggctt caggttcagc agggtggaca tgagctgggt gaggcaggcc 120
cccggcaagg gcctggagtg ggtggccagg atcaacgacg acggcagctg gagggacgcc 180
gccgacagcg tgaagggcag gttcaccatc agcagggaca acgccaagaa caccctgtac 240
ctgcagatga acagcctgaa gaccgaggac accgccgtgt actactgcgc caccggcttc 300
cccagcggcg gcaactggat ccagaggggc cagggcaccc aggtgaccgt gagcagc 357
<210> 65
<211> 384
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 65
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
agctgcgccg tgagcggcga caccgccaac agcaactgca tgggctggtt caggcaggcc 120
cccggcgagg agagggagtg ggtggccagc atcgtgagcg ccagcggctt catctactac 180
gccgacgccg tgaggggcag gttcaccgtg agccaggaca agagcaagaa caccgtgtac 240
ctgcagatga acagcctgaa gcccgaggac accgccatgt actactgcgc cgccaagccc 300
aggcccaggt tcctgaccgt gcccgccggc tgcggctacg agtacaacta ctggggccag 360
ggcacccagg tgaccgtgag cagc 384
<210> 66
<211> 375
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 66
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
agctgcgccg ccagcggcag cacctacacc atcgcctggt tcaggcaggc ccccggcaag 120
gagagggagg gcatcgccgc catctacacc ctggccggcg gcacctacta cgccgacagc 180
gtgaagggca ggttcaccat cagcagggac aacgccaaga acaccctgag cctgcagatg 240
aacagcctga agcccgagga caccgccatg tactactgcg ccgccgccat cggcgccggc 300
tacggcagga actggctgct gagcgagagg tggtacaact actggggcca gggcacccag 360
gtgaccgtga gcagc 375
<210> 67
<211> 381
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 67
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
agctgcgccg ccagcggcta cacctacagc agcagctgca tgggctggtt caggcaggcc 120
cccggcaagg agagggaggg cgtggccacc atcgacagct tcggcaacac caagtacgcc 180
gacagcgtga agggcaggtt caccatcagc aaggacaacg ccaagaacac cctgtacctg 240
cagatgaaca gcctgaagcc cgaggacacc gccatgtact actgcaccgc cgacatggac 300
ggcaggtggg gcggctactg cgtgaggagc gccgaggact tcgcctactg gggccagggc 360
acccaggtga ccgtgagcag c 381
<210> 68
<211> 363
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 68
caggtgcagc tggtggagag cggcggcggc agcgtgcaga ccggcggcag cctgaggctg 60
agctgcgccg ccagcggcca cggcagctgc acctacgaca tgagctggta caggcaggcc 120
cccggcaaga gcagggagtt cgtgagcgtg atcatgagcg acggcaccac cgcctacacc 180
agcagcgtga aggacaggtt caccatcagc caggacaacg ccaagagcac cgtgtacctg 240
cagatgaaca gcctgaagcc cgaggacacc gccaggtact actgcgaggc cgtgctgagg 300
ggcaggtgca ccaggagcgg cttcacctac tggggccagg gcacccaggt gaccgtgagc 360
agc 363
<210> 69
<211> 378
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 69
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
agctgcaccg ccagcggcag cacctacagc aggaggtaca tcggctggtt caggcaggcc 120
cccggcaagg agagggaggg cgtggccacc atcagcaccg gcggcggctt caggagctac 180
gccgacagcg tgaagggcag gttcaccatc agccaggaca acgccaagaa caccgtgtac 240
ctgcagatga acagcctgaa gcccgaggac accgccatgt actactgcgc cgccggcagc 300
aacgtggaca ggaggtggag cctgctgagc agggactaca actactgggg ccagggcacc 360
caggtgaccg tgagcagc 378
<210> 70
<211> 360
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 70
caggtgcagc tggtggagag cggcggcggc agcgtgcagc ccggcggcag cctgaggctg 60
agctgcgtgg tgagcggcta caccatcgac aggctgtgca ccggctggtt caggcaggcc 120
cccggcaagg agagggagtt cgtgagcagc accagcaccg acggcatcac caggtacgcc 180
gacagcgtga agggcaggtt caccgtgagc agcgacaagg ccaaggacac cgtgtacctg 240
cagatgaaca gcctgaagcc cgaggacacc gccatgtact tctgccactt cggcggcagg 300
tggctgaccc ccagggcctg cgactactgg ggccagggca cccaggtgac cgtgagcagc 360
<210> 71
<211> 363
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 71
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
agctgcgccg ccagcggcgc cgcctacagc gactactgca tgggctggtt caggcaggcc 120
cccggcaagg agagggagag ggtggccacc ttcctgaggg acggcagcgc cgactacgcc 180
gccagcgtga ccggcaggtt caccatcagc aaggacaacg tgaagaacac cctgtacctg 240
cagatgaaca gcctgaagcc cgaggacacc gccatgtact actgcgccgc cgactactgg 300
ggctaccagt acaactgcag gttcgagtac tggggccagg gcacccaggt gaccgtgagc 360
agc 363
<210> 72
<211> 396
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 72
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
agctgcgccg tgagcggcta caccgtgagc aggagctgca tggcctggtt caggcaggtg 120
cccggcaagg agagggaggg cgtggccagc atcagcagcg acggcggcat caccacctac 180
gtggacagcg tgaagggcag gttcaccatc agccaggaca acgccaagaa caccctgtac 240
ctgcagatga acagcgtgaa gcccgaggac accgccgtgt actactgcgc caccggcgtg 300
ctgggcttct tcagcaacct gtgcaggagc accgtggtgc cccccaccgc caggttcagg 360
tactggggcc agggcaccca ggtgaccgtg agcagc 396
<210> 73
<211> 363
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 73
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
agctgcgccg ccagcggcgc cgcctacagc gactactgca tgggctggtt caggcaggcc 120
cccggcaagg agagggagag ggtggccacc ttcctgaggg acggcagcgc cgactacgcc 180
gccagcgtga ccggcaggtt caccatcagc aaggacaacg tgaagaacac cctgtacctg 240
cagatgaaca gcctgaagcc cgaggacacc gccatgtact actgcgccgt ggactactgg 300
ggctacggct acaactgcag gttcgagtac tggggccagg gcacccaggt gaccgtgagc 360
agc 363
<210> 74
<211> 381
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 74
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
acctgcgccg tgagcggcta cagctacagc agcaaccaca tgggctggtt caggcaggcc 120
cccggcaagg agagggaggg cgtggccgtg atcttcaccg gcggcggcag gacctggtac 180
gccaacagcg tgaagggcag gttcaccatc agccacgaca acgccaagaa caccgtgtac 240
ctgcagatga acagcctgaa gctggaggac accgccatgt actactgcgc cgccgactac 300
gtgcccgtgg gcaggaggta cgccgtgctg gaccccgact acacctactg gggccagggc 360
acccaggtga ccgtgagcag c 381
<210> 75
<211> 375
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 75
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
agctgcgccg ccagcggcta cacctggagc agcacctgca tgggctggtt caggcaggcc 120
cccggcaagg agaggcaggg cgtggccgcc atcagccccg gcggcagctt caagtactac 180
gccgacagcg tgaagggcag gttcaccatc agcagggaca acgccaagaa caccgtgtac 240
ctgcagatga acagcctgca gcccgaggac accgccatgt actactgcgc cgccgccccc 300
ttcgcctgcg acggcgactg gtacagcggc aggtggaaca actggggcca gggcacccag 360
gtgaccgtga gcagc 375
<210> 76
<211> 378
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 76
caggtgcagc tggtggagag cggcggcggc ctggtgcagc ccggcggcag ccccaagctg 60
agctgcgccg ccagcggctt caccttcagc aactacgcca tgagctgggt gaggcaggcc 120
cccggcaagg tgctggagtg ggtgagcttc atcaacagcg gcggcggcgg caccgcctac 180
gccgaccccg tgaagggcag gttcaccatc agcagggaca acgccaagaa caccctgtac 240
ctgcagctga acagcctgaa gaccgaggac accgccatgt actactgcgc caagggcctg 300
cagtacgaca cctactgcag cagccagtac tgctacgcca accccggcgg ccagggcacc 360
caggtgaccg tgagcagc 378
<210> 77
<211> 378
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 77
caggtgcagc tggtggagag cggcggcggc accgtgcagg ccggcggcag cctgaggctg 60
agctgcgccg ccagcggcta cacctacagc agcgcctgca tgggctggtt caggcaggcc 120
cccggcaagg agagggaggg cgtggccacc atcggcagcg acggcagcac cagctacacc 180
gacagcgtga agggcaggtt caccatcagc aaggacaacg tgcacaacaa cctgtacctg 240
cagatgaaca gcctgaagcc cgaggacacc gccatgtact actgcgccgc caccatcacc 300
accgtgacca tgtgcaggag gagggacggc aggctgttcg gccactgggg ccagggcacc 360
caggtgaccg tgagcagc 378
<210> 78
<211> 378
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 78
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
agctgcgccg tgagcggctt cccctacagc acctactaca tgggctggtt caggcaggcc 120
cccggcaagg agagggaggg cgtggccgcc gtgtaccccg gcgtgggcct gaagaggtac 180
gccgacagcg tgaagggcag gttcaccatc agccaggacg acgccaagaa caccctgtac 240
ctgcagatga acagcctgaa gcccgaggac accgccctgt actactgcgc cgtggaccag 300
gccgtgagcc tgggctggga gctgagcccc ggcaggtaca ggtactgggg ccagggcacc 360
caggtgaccg tgagcagc 378
<210> 79
<211> 375
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 79
caggtgcagc tggtggagag cggcggcggc agcgtgcaga ccggcggcag cctgaagctg 60
agctgcgtgc acagcggcta cagctacagc agcgtgtgca tggcctggtt caggcaggcc 120
cccggcaagg agagggaggg cgtggccgcc atcagccccg gcggcagctt caagtactac 180
gtggacagcg tgaagggcag gttcaccatc agcagggaca acagcaagaa caccggctac 240
ctgcagatga acagcctgaa gcccgaggac accgccatgt actactgcgc caccagcagg 300
agcggctgca gggacgactt cttccagtac gagtacgact actggggcca gggcacccag 360
gtgaccgtga gcagc 375
<210> 80
<211> 375
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 80
caggtgcagc tggtggagag cggcggcggc agcgcccagg ccggcggcag cctgaggctg 60
agctgcgccg ccagcggcaa cagctacagc aggaactgca tgggctggtt caggcaggcc 120
cccggcaagg agaggcaggg cgtggccgcc atcagccccg gcggcagctt caagtactac 180
gccgacagcg tgaagggcag gttcaccatc agcagggaca acgccaagaa caccgtgtac 240
ctgcagatga acagcctgca gcccgaggac accgccatgt actactgcgc cgccgccccc 300
ctggcctgcg acggcgactg gtacagcggc aggtggaaca actggggcca gggcacccag 360
gtgaccgtga gcagc 375
<210> 81
<211> 384
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 81
caggtgcagc tggtggagag cggcggcggc agcgtgcccg ccggcggcag cctgaggctg 60
agctgcgccg ccagcggcta cacctacagc ggcagctgca tgggctggtt caggcaggcc 120
cccggcaagg agagggagga ggtggccacc atctacaccg acaacggctt caagtactac 180
accgagagcg tgaagggcag gttcaccatc agccaggaca aggccaagaa caccgtgtac 240
ctgcagatga acagcctgaa gcccgaggac accgccgtgt actactgcgc cgccgacggc 300
ggcaccggcc tgtggtacac ctgcgccagg ggccagatca gccacaacta ctggggccag 360
ggcacccagg tgaccgtgag cagc 384
<210> 82
<211> 381
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 82
caggtgcagc tggtggagag cggcggcggc agcgtgcagg ccggcggcag cctgaggctg 60
agctgcaccg tgagcggcta cagctacagc agcgactaca tgggctggtt caggcaggcc 120
cccggcaagg agagggaggg cgtggccgtg atcttcaccg gcggcggcag gacctggtac 180
gccaacagcg tgaagggcag gttcaccatc agccaggaca acgccaagaa caccgtgtac 240
ctgcagatga acagcctgaa gctggaggac accgccatgt actactgcgc cgccgactac 300
gtgcccgtgg gcaggaggta cgccgtgctg gaccccgact acacctactg gggccagggc 360
acccaggtga ccgtgagcag c 381
<210> 83
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 83
Gly Tyr Ser Gly Arg Ser Asp Tyr
1 5
<210> 84
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 84
Gly Ala Ala Tyr Ser Asp Tyr Cys
1 5
<210> 85
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 85
Gly Tyr Asp Tyr Ser Thr Tyr Cys
1 5
<210> 86
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 86
Gly Phe Thr Phe Ser Ser Tyr Pro
1 5
<210> 87
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 87
Gly Phe Thr Phe Ser Arg Tyr Asp
1 5
<210> 88
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 88
Gly Tyr Ile Ser His Pro Tyr Cys
1 5
<210> 89
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 89
Gly Tyr Thr Tyr Ser Arg Tyr Ser
1 5
<210> 90
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 90
Gly Tyr Ala Tyr Ser Arg Tyr Ser
1 5
<210> 91
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 91
Gly Val Thr Ser Lys Arg Ala Cys
1 5
<210> 92
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 92
Gly Asn Ser Tyr Ser Arg Asn Cys
1 5
<210> 93
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 93
Gly Tyr Thr His Ser Ser Tyr Cys
1 5
<210> 94
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 94
Gly Tyr Thr Tyr Ser Ser Asn Cys
1 5
<210> 95
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 95
Gly Tyr Ala Tyr Ser Arg Tyr Tyr
1 5
<210> 96
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 96
Gly Tyr Thr Ile Ser Ser Leu Cys
1 5
<210> 97
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 97
Gly Asp Thr Asn Arg Gln Arg Cys
1 5
<210> 98
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 98
Gly Tyr Thr Val Asn Thr Lys Cys
1 5
<210> 99
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 99
Gly Tyr Thr Tyr Ser Thr Asn Cys
1 5
<210> 100
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 100
Gly Tyr Thr Gly Asp Thr Ser Ala
1 5
<210> 101
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 101
Gly Tyr Thr Tyr Ser Ser Ala Cys
1 5
<210> 102
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 102
Gly Tyr Thr Tyr Ser Tyr Asn Cys
1 5
<210> 103
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 103
Gly Phe Arg Phe Ser Arg Val Asp
1 5
<210> 104
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 104
Gly Asp Thr Ala Asn Ser Asn Cys
1 5
<210> 105
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 105
Gly Ser Thr Tyr Thr Ile Ala Trp
1 5
<210> 106
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 106
Gly Tyr Thr Tyr Ser Ser Ser Cys
1 5
<210> 107
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 107
Gly His Gly Ser Cys Thr Tyr Asp
1 5
<210> 108
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 108
Gly Ser Thr Tyr Ser Arg Arg Tyr
1 5
<210> 109
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 109
Gly Tyr Thr Ile Asp Arg Leu Cys
1 5
<210> 110
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 110
Gly Tyr Thr Val Ser Arg Ser Cys
1 5
<210> 111
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 111
Gly Tyr Ser Tyr Ser Ser Asn His
1 5
<210> 112
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 112
Gly Tyr Thr Trp Ser Ser Thr Cys
1 5
<210> 113
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 113
Gly Phe Thr Phe Ser Asn Tyr Ala
1 5
<210> 114
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 114
Gly Phe Pro Tyr Ser Thr Tyr Tyr
1 5
<210> 115
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 115
Gly Tyr Ser Tyr Ser Ser Val Cys
1 5
<210> 116
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 116
Gly Tyr Thr Tyr Ser Gly Ser Cys
1 5
<210> 117
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 117
Gly Tyr Ser Tyr Ser Ser Asp Tyr
1 5
<210> 118
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 118
Ile Phe Thr Gly Gly Gly Arg Thr
1 5
<210> 119
<211> 7
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 119
Phe Leu Arg Asp Gly Ser Ala
1 5
<210> 120
<211> 7
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 120
Ile Ser Thr Val Gly Thr Thr
1 5
<210> 121
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 121
Ile Asn Ser Gly Gly Gly Ser Thr
1 5
<210> 122
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 122
Ile Tyr Ser Gly Ser Gly Asn Thr
1 5
<210> 123
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 123
Ile Ser Ile Gly Gly Gly Gly Thr
1 5
<210> 124
<211> 7
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 124
Leu Asp Ser Asp Gly Ala Thr
1 5
<210> 125
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 125
Ile Ser Arg Pro Gly Gly Ala Thr
1 5
<210> 126
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 126
Ile Ser Pro Gly Gly Ser Phe Lys
1 5
<210> 127
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 127
Ile Tyr Thr Gly Gly Gly Ser Thr
1 5
<210> 128
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 128
Ile Ser Thr Gly Gly Gly Thr Thr
1 5
<210> 129
<211> 7
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 129
Ile Asp Arg Asp Gly Gly Thr
1 5
<210> 130
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 130
Ile Tyr Thr Gly Thr Ser Thr Thr
1 5
<210> 131
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 131
Ile Tyr Arg Val Ser Ser Ala Thr
1 5
<210> 132
<211> 7
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 132
Ile Gly Ser Asp Gly Ser Thr
1 5
<210> 133
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 133
Ile Asn Asp Asp Gly Ser Trp Arg
1 5
<210> 134
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 134
Ile Val Ser Ala Ser Gly Phe Ile
1 5
<210> 135
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 135
Ile Tyr Thr Leu Ala Gly Gly Thr
1 5
<210> 136
<211> 7
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 136
Ile Asp Ser Phe Gly Asn Thr
1 5
<210> 137
<211> 7
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 137
Ile Met Ser Asp Gly Thr Thr
1 5
<210> 138
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 138
Ile Ser Thr Gly Gly Gly Phe Arg
1 5
<210> 139
<211> 7
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 139
Thr Ser Thr Asp Gly Ile Thr
1 5
<210> 140
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 140
Ile Ser Ser Asp Gly Gly Ile Thr
1 5
<210> 141
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 141
Ile Asn Ser Gly Gly Gly Gly Thr
1 5
<210> 142
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 142
Val Tyr Pro Gly Val Gly Leu Lys
1 5
<210> 143
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 143
Ile Tyr Thr Asp Asn Gly Phe Lys
1 5
<210> 144
<211> 21
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 144
Ala Ala Asp Tyr Val Pro Val Gly Arg Arg Tyr Ala Val Leu Asp Pro
1 5 10 15
Asp Tyr Thr Tyr Trp
20
<210> 145
<211> 16
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 145
Ala Ala Asp Tyr Trp Gly Tyr Gly Tyr Asn Cys Arg Phe Glu Tyr Trp
1 5 10 15
<210> 146
<211> 22
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 146
Ala Ser Leu Asp Arg Arg Met Tyr Gly Gly Ile Trp Tyr Cys Asp Leu
1 5 10 15
Arg Arg Tyr Asn Tyr Trp
20
<210> 147
<211> 9
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 147
Ala Leu Asp Ser Arg Gly Thr Gln Arg
1 5
<210> 148
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 148
Ala Ala Asp Trp Thr Gly Arg Thr Leu His Glu Tyr Asn Tyr Leu
1 5 10 15
<210> 149
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 149
Ala Ala Arg Gly Gly Tyr Tyr Cys Thr Ala Ser Ala Ser Tyr Asn Tyr
1 5 10 15
Trp
<210> 150
<211> 21
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 150
Ala Ala Gln Arg Ile Arg Trp Val Ala Phe Val Ser Leu Arg Ser Ala
1 5 10 15
Asp Phe Asp Tyr Trp
20
<210> 151
<211> 23
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 151
Ala Phe Arg Val Ile Asp Pro Phe Asp Thr Cys Ser Ser Arg Arg Arg
1 5 10 15
Asp Gly Asp Phe Pro Tyr Trp
20
<210> 152
<211> 19
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 152
Ala Ala Ala Pro Phe Ala Cys Asp Gly Asp Trp Tyr Ser Gly Arg Trp
1 5 10 15
Asn Asn Trp
<210> 153
<211> 19
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 153
Ala Ala Ala Pro Phe Ala Cys Asp Gly Asp Trp His Ser Gly Arg Trp
1 5 10 15
Asn Asn Trp
<210> 154
<211> 24
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 154
Ala Ala Asp Leu Arg Ala Ala Pro Tyr Gly Gly Ile Gly Tyr Phe Pro
1 5 10 15
Ala Ala Thr Asp Phe Gly Tyr Trp
20
<210> 155
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 155
Ala Ala Arg Val Gly Ser Pro Phe Gly Gly Ala Pro Asp Leu Ser Ser
1 5 10 15
Leu Arg Pro Glu Arg Tyr Ser Tyr Trp
20 25
<210> 156
<211> 16
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 156
Ala Ala Asp Tyr Trp Gly Tyr Gln Tyr Asn Cys Arg Phe Glu Tyr Trp
1 5 10 15
<210> 157
<211> 21
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 157
Ala Ala Thr Thr Tyr Ile Gly Ser Trp Cys Gly Leu Arg Ser Pro Leu
1 5 10 15
Asp Tyr Asn Asn Trp
20
<210> 158
<211> 19
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 158
Ala Ala Ala Pro Phe Ala Cys Asp Gly Asp Trp His Ser Arg Arg Trp
1 5 10 15
Asn Asn Trp
<210> 159
<211> 20
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 159
Ala Ala Arg Ser Thr Ala Val Thr Leu Arg Gln Leu Thr Gln Gln Ala
1 5 10 15
Tyr Asn Tyr Trp
20
<210> 160
<211> 21
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 160
Ala Ala Thr Ile Thr Thr Val Thr Met Cys Arg Arg Arg Asp Gly Arg
1 5 10 15
Leu Phe Gly His Trp
20
<210> 161
<211> 13
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 161
Ala Thr Gly Phe Pro Ser Gly Gly Asn Trp Ile Gln Arg
1 5 10
<210> 162
<211> 22
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 162
Ala Ala Lys Pro Arg Pro Arg Phe Leu Thr Val Pro Ala Gly Cys Gly
1 5 10 15
Tyr Glu Tyr Asn Tyr Trp
20
<210> 163
<211> 22
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 163
Ala Ala Ala Ile Gly Ala Gly Tyr Gly Arg Asn Trp Leu Leu Ser Glu
1 5 10 15
Arg Trp Tyr Asn Tyr Trp
20
<210> 164
<211> 22
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 164
Thr Ala Asp Met Asp Gly Arg Trp Gly Gly Tyr Cys Val Arg Ser Ala
1 5 10 15
Glu Asp Phe Ala Tyr Trp
20
<210> 165
<211> 16
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 165
Glu Ala Val Leu Arg Gly Arg Cys Thr Arg Ser Gly Phe Thr Tyr Trp
1 5 10 15
<210> 166
<211> 20
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 166
Ala Ala Gly Ser Asn Val Asp Arg Arg Trp Ser Leu Leu Ser Arg Asp
1 5 10 15
Tyr Asn Tyr Trp
20
<210> 167
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 167
His Phe Gly Gly Arg Trp Leu Thr Pro Arg Ala Cys Asp Tyr Trp
1 5 10 15
<210> 168
<211> 26
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 168
Ala Thr Gly Val Leu Gly Phe Phe Ser Asn Leu Cys Arg Ser Thr Val
1 5 10 15
Val Pro Pro Thr Ala Arg Phe Arg Tyr Trp
20 25
<210> 169
<211> 16
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 169
Ala Val Asp Tyr Trp Gly Tyr Gly Tyr Asn Cys Arg Phe Glu Tyr Trp
1 5 10 15
<210> 170
<211> 20
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 170
Ala Lys Gly Leu Gln Tyr Asp Thr Tyr Cys Ser Ser Gln Tyr Cys Tyr
1 5 10 15
Ala Asn Pro Gly
20
<210> 171
<211> 20
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 171
Ala Val Asp Gln Ala Val Ser Leu Gly Trp Glu Leu Ser Pro Gly Arg
1 5 10 15
Tyr Arg Tyr Trp
20
<210> 172
<211> 19
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 172
Ala Thr Ser Arg Ser Gly Cys Arg Asp Asp Phe Phe Gln Tyr Glu Tyr
1 5 10 15
Asp Tyr Trp
<210> 173
<211> 19
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 173
Ala Ala Ala Pro Leu Ala Cys Asp Gly Asp Trp Tyr Ser Gly Arg Trp
1 5 10 15
Asn Asn Trp
<210> 174
<211> 22
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 174
Ala Ala Asp Gly Gly Thr Gly Leu Trp Tyr Thr Cys Ala Arg Gly Gln
1 5 10 15
Ile Ser His Asn Tyr Trp
20
<210> 175
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 175
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Val Ser
20 25
<210> 176
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 176
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser
20 25
<210> 177
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 177
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser
20 25
<210> 178
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 178
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser
20 25
<210> 179
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 179
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Trp Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser
20 25
<210> 180
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 180
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Ala Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser
20 25
<210> 181
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 181
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ala Cys Ala Ala Ser
20 25
<210> 182
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 182
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Gly Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Val Val Ser
20 25
<210> 183
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 183
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Thr Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser
20 25
<210> 184
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 184
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Val Ala Ser
20 25
<210> 185
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 185
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Pro
20 25
<210> 186
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 186
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ser Val Ser
20 25
<210> 187
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 187
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ala
20 25
<210> 188
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 188
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Thr Ala Ser
20 25
<210> 189
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 189
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Val Val Ser
20 25
<210> 190
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 190
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Thr Cys Ala Val Ser
20 25
<210> 191
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 191
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Pro Lys Leu Ser Cys Ala Ala Ser
20 25
<210> 192
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 192
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Thr Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser
20 25
<210> 193
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 193
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Thr Gly Gly
1 5 10 15
Ser Leu Lys Leu Ser Cys Val His Ser
20 25
<210> 194
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 194
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Pro Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser
20 25
<210> 195
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 195
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Thr Val Ser
20 25
<210> 196
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 196
Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val Ala
1 5 10 15
Val
<210> 197
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 197
Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Arg Val Ala
1 5 10 15
Thr
<210> 198
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 198
Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val Ala
1 5 10 15
Ser
<210> 199
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 199
Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser
1 5 10 15
Ala
<210> 200
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 200
Met Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Val Ser
1 5 10 15
Ala
<210> 201
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 201
Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val Ala
1 5 10 15
Gly
<210> 202
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 202
Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Trp Val Ala
1 5 10 15
Thr
<210> 203
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 203
Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Gln Gly Val Ala
1 5 10 15
Ala
<210> 204
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 204
Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val Ala
1 5 10 15
Val
<210> 205
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 205
Met Gly Trp Phe Arg Gln Val Pro Gly Lys Glu Arg Glu Gly Val Ala
1 5 10 15
Thr
<210> 206
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 206
Met Gly Trp Phe Arg Arg Thr Glu Arg Glu Ala Val Ala Arg
1 5 10
<210> 207
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 207
Met Gly Trp Phe Arg Gln Ala Pro Gly Glu Gly Arg Glu Gly Val Ala
1 5 10 15
Ala
<210> 208
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 208
Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val Ala
1 5 10 15
Thr
<210> 209
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 209
Met Gly Trp Leu Arg Gln Ala Pro Gly Lys Glu Arg Gln Gly Val Ala
1 5 10 15
Ala
<210> 210
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 210
Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val Ala
1 5 10 15
Ser
<210> 211
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 211
Val Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Gln Gly Val Ala
1 5 10 15
Ala
<210> 212
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 212
Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala
1 5 10 15
Arg
<210> 213
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 213
Met Gly Trp Phe Arg Gln Ala Pro Gly Glu Glu Arg Glu Trp Val Ala
1 5 10 15
Ser
<210> 214
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 214
Ile Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Ile Ala
1 5 10 15
Ala
<210> 215
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 215
Met Ser Trp Tyr Arg Gln Ala Pro Gly Lys Ser Arg Glu Phe Val Ser
1 5 10 15
Val
<210> 216
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 216
Ile Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val Ala
1 5 10 15
Thr
<210> 217
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 217
Thr Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val Ser
1 5 10 15
Ser
<210> 218
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 218
Met Ala Trp Phe Arg Gln Val Pro Gly Lys Glu Arg Glu Gly Val Ala
1 5 10 15
Ser
<210> 219
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 219
Met Ser Trp Val Arg Gln Ala Pro Gly Lys Val Leu Glu Trp Val Ser
1 5 10 15
Phe
<210> 220
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 220
Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val Ala
1 5 10 15
Ala
<210> 221
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 221
Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val Ala
1 5 10 15
Ala
<210> 222
<211> 17
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 222
Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Glu Val Ala
1 5 10 15
Thr
<210> 223
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 223
Trp Tyr Ala Asn Ser Val Lys Gly Arg Phe Thr Ile Ser Gln Gly Asn
1 5 10 15
Ala Glu Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Leu Glu Asp
20 25 30
Thr Ala Met Tyr Tyr Cys
35
<210> 224
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 224
Asp Tyr Ala Ala Ser Val Thr Gly Arg Phe Thr Ile Ser Lys Asp Asn
1 5 10 15
Val Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp
20 25 30
Thr Ala Met Tyr Tyr Cys
35
<210> 225
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 225
Ser Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn
1 5 10 15
Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp
20 25 30
Thr Asn Met Tyr Tyr Cys
35
<210> 226
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 226
Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn
1 5 10 15
Ala Lys Asn Thr Leu Tyr Leu Gln Leu Asn Gly Leu Lys Thr Glu Asp
20 25 30
Thr Ala Met Tyr Tyr Cys
35
<210> 227
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 227
Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn
1 5 10 15
Ala Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp
20 25 30
Thr Ala Val Tyr Tyr Cys
35
<210> 228
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 228
Trp Tyr Ala Asn Ser Val Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn
1 5 10 15
Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Leu Glu Asp
20 25 30
Thr Ala Met Tyr Tyr Cys
35
<210> 229
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 229
Phe Thr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn
1 5 10 15
Ala Lys Asn Thr Val His Leu Arg Met Asn Ser Leu Lys Pro Glu Asp
20 25 30
Thr Ala Met Tyr Tyr Cys
35
<210> 230
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 230
Ile Tyr Thr Asp Ser Val Lys Gly Arg Phe Thr Ile Ala Lys Asp Asn
1 5 10 15
Ala Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp
20 25 30
Thr Gly Met Tyr Tyr Cys
35
<210> 231
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 231
Tyr Ser Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn
1 5 10 15
Ala Glu Asn Met Ala Phe Leu Ile Leu Asn Ser Leu Lys Ala Glu Asp
20 25 30
Ala Ala Ile Tyr Tyr Cys
35
<210> 232
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 232
Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn
1 5 10 15
Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Gln Pro Glu Asp
20 25 30
Thr Ala Met Tyr Tyr Cys
35
<210> 233
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 233
Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn
1 5 10 15
Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp
20 25 30
Thr Ala Met Tyr Tyr Cys
35
<210> 234
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 234
Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Val Ser Arg Asp Asn
1 5 10 15
Ala Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Thr Pro Glu Asp
20 25 30
Thr Ala Ile Tyr Tyr Cys
35
<210> 235
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 235
Arg Tyr Ala Asp Pro Val Lys Gly Arg Phe Thr Ile Ser Lys Asp Gly
1 5 10 15
Ala Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp
20 25 30
Thr Ala Met Tyr Tyr Cys
35
<210> 236
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 236
Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn
1 5 10 15
Ala Arg Asn Thr Ile Tyr Leu Gln Met Asp Ser Val Lys Pro Asp Asp
20 25 30
Thr Ala Met Tyr Tyr Cys
35
<210> 237
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 237
Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn
1 5 10 15
Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp
20 25 30
Thr Ala Met Tyr Tyr Cys
35
<210> 238
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 238
Phe Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Gln Asp Lys
1 5 10 15
Ala Asn Asp Ser Leu Tyr Leu Gln Met Asn Asn Leu Lys Pro Glu Asp
20 25 30
Thr Ala Val Tyr Tyr Cys
35
<210> 239
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 239
Ser Tyr Thr Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Lys Asp Asn
1 5 10 15
Val His Asn Asn Leu Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp
20 25 30
Thr Ala Met Tyr Tyr Cys
35
<210> 240
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 240
Asp Ala Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn
1 5 10 15
Ala Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp
20 25 30
Thr Ala Val Tyr Tyr Cys
35
<210> 241
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 241
Tyr Tyr Ala Asp Ala Val Arg Gly Arg Phe Thr Val Ser Gln Asp Lys
1 5 10 15
Ser Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp
20 25 30
Thr Ala Met Tyr Tyr Cys
35
<210> 242
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 242
Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn
1 5 10 15
Ala Lys Asn Thr Leu Ser Leu Gln Met Asn Ser Leu Lys Pro Glu Asp
20 25 30
Thr Ala Met Tyr Tyr Cys
35
<210> 243
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 243
Lys Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Lys Asp Asn
1 5 10 15
Ala Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp
20 25 30
Thr Ala Met Tyr Tyr Cys
35
<210> 244
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 244
Ala Tyr Thr Ser Ser Val Lys Asp Arg Phe Thr Ile Ser Gln Asp Asn
1 5 10 15
Ala Lys Ser Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp
20 25 30
Thr Ala Arg Tyr Tyr Cys
35
<210> 245
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 245
Ser Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn
1 5 10 15
Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp
20 25 30
Thr Ala Met Tyr Tyr Cys
35
<210> 246
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 246
Arg Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Val Ser Ser Asp Lys
1 5 10 15
Ala Lys Asp Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp
20 25 30
Thr Ala Met Tyr Phe Cys
35
<210> 247
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 247
Thr Tyr Val Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn
1 5 10 15
Ala Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Val Lys Pro Glu Asp
20 25 30
Thr Ala Val Tyr Tyr Cys
35
<210> 248
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 248
Trp Tyr Ala Asn Ser Val Lys Gly Arg Phe Thr Ile Ser His Asp Asn
1 5 10 15
Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Leu Glu Asp
20 25 30
Thr Ala Met Tyr Tyr Cys
35
<210> 249
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 249
Ala Tyr Ala Asp Pro Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn
1 5 10 15
Ala Lys Asn Thr Leu Tyr Leu Gln Leu Asn Ser Leu Lys Thr Glu Asp
20 25 30
Thr Ala Met Tyr Tyr Cys
35
<210> 250
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 250
Arg Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Gln Asp Asp
1 5 10 15
Ala Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp
20 25 30
Thr Ala Leu Tyr Tyr Cys
35
<210> 251
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 251
Tyr Tyr Val Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn
1 5 10 15
Ser Lys Asn Thr Gly Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp
20 25 30
Thr Ala Met Tyr Tyr Cys
35
<210> 252
<211> 38
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 252
Tyr Tyr Thr Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Gln Asp Lys
1 5 10 15
Ala Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp
20 25 30
Thr Ala Val Tyr Tyr Cys
35
<210> 253
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 253
Gly Gln Gly Thr Gln Val Thr Val Ser Ser
1 5 10
<210> 254
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 254
Gly Arg Gly Thr Gln Val Thr Val Ser Ser
1 5 10
<210> 255
<211> 13
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 255
ggatcgaacc ctt 13
<210> 256
<211> 21
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 256
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Asp
20

Claims (10)

1.针对甲型肝炎病毒的单域抗体, 其特征在于，所述的单域抗体的序列包括互补决定区CDR；所述互补决定区CDR包括CDR1、CDR2和CDR3的氨基酸序列；所述的CDR1的氨基酸序列如SEQ ID NO:83- SEQ ID NO:117中的任意一条所示；所述的CDR2的氨基酸序列如SEQ IDNO:118-SEQ ID NO:143中的任意一条所示；所述的CDR3的氨基酸序列如SEQ ID NO:144-SEQ ID NO:174中的任意一条所示。
2.针对甲型肝炎病毒的单域抗体, 其特征在于，所述的单域抗体的序列包括互补决定区CDR；所述互补决定区CDR包括CDR1、CDR2和CDR3的氨基酸序列；所述的单域抗体的互补决定区CDR的序列为下述（1）-（39）中的一种：
（1）SEQ ID NO:83所示的CDR1，SEQ ID NO:118所示的CDR2，SEQ ID NO:144所示的CDR3；
（2）SEQ ID NO:84所示的CDR1，SEQ ID NO:119所示的CDR2，SEQ ID NO:145所示的CDR3；
（3）SEQ ID NO:84所示的CDR1，SEQ ID NO:119所示的CDR2，SEQ ID NO:156所示的CDR3；
（4）SEQ ID NO:84所示的CDR1，SEQ ID NO:119所示的CDR2，SEQ ID NO:169所示的CDR3；
（5）SEQ ID NO:85所示的CDR1，SEQ ID NO:120所示的CDR2，SEQ ID NO:146所示的CDR3；
（6）SEQ ID NO:86所示的CDR1，SEQ ID NO:121所示的CDR2，SEQ ID NO:147所示的CDR3；
（7）SEQ ID NO:87所示的CDR1，SEQ ID NO:122所示的CDR2，SEQ ID NO:148所示的CDR3；
（8）SEQ ID NO:88所示的CDR1，SEQ ID NO:123所示的CDR2，SEQ ID NO:149所示的CDR3；
（9）SEQ ID NO:89所示的CDR1，SEQ ID NO:124所示的CDR2，SEQ ID NO:150所示的CDR3；
（10）SEQ ID NO:90所示的CDR1，SEQ ID NO:124所示的CDR2，SEQ ID NO:150所示的CDR3；
（11）SEQ ID NO:91所示的CDR1，SEQ ID NO:125所示的CDR2，SEQ ID NO:151所示的CDR3；
（12）SEQ ID NO:92所示的CDR1，SEQ ID NO:126所示的CDR2，SEQ ID NO:152所示的CDR3；
（13）SEQ ID NO:92所示的CDR1，SEQ ID NO:126所示的CDR2，SEQ ID NO:153所示的CDR3；
（14）SEQ ID NO:92所示的CDR1，SEQ ID NO:126所示的CDR2，SEQ ID NO:173所示的CDR3；
（15）SEQ ID NO:93所示的CDR1，SEQ ID NO:126所示的CDR2，SEQ ID NO:152所示的CDR3；
（16）SEQ ID NO:94所示的CDR1，SEQ ID NO:127所示的CDR2，SEQ ID NO:154所示的CDR3；
（17）SEQ ID NO:95所示的CDR1，SEQ ID NO:128所示的CDR2，SEQ ID NO:155所示的CDR3；
（18）SEQ ID NO:96所示的CDR1，SEQ ID NO:129所示的CDR2，SEQ ID NO:156所示的CDR3；
（19）SEQ ID NO:97所示的CDR1，SEQ ID NO:126所示的CDR2，SEQ ID NO:152所示的CDR3；
（20）SEQ ID NO:98所示的CDR1，SEQ ID NO:130所示的CDR2，SEQ ID NO:157所示的CDR3；
（21）SEQ ID NO:99所示的CDR1，SEQ ID NO:126所示的CDR2，SEQ ID NO:158所示的CDR3；
（22）SEQ ID NO:100所示的CDR1，SEQ ID NO:131所示的CDR2，SEQ ID NO:159所示的CDR3；
（23）SEQ ID NO:101所示的CDR1，SEQ ID NO:132所示的CDR2，SEQ ID NO:160所示的CDR3；
（24）SEQ ID NO:102所示的CDR1，SEQ ID NO:126所示的CDR2，SEQ ID NO:152所示的CDR3；
（25）SEQ ID NO:103所示的CDR1，SEQ ID NO:133所示的CDR2，SEQ ID NO:161所示的CDR3；
（26）SEQ ID NO:104所示的CDR1，SEQ ID NO:134所示的CDR2，SEQ ID NO:162所示的CDR3；
（27）SEQ ID NO:105所示的CDR1，SEQ ID NO:135所示的CDR2，SEQ ID NO:163所示的CDR3；
（28）SEQ ID NO:106所示的CDR1，SEQ ID NO:136所示的CDR2，SEQ ID NO:164所示的CDR3；
（29）SEQ ID NO:107所示的CDR1，SEQ ID NO:137所示的CDR2，SEQ ID NO:165所示的CDR3；
（30）SEQ ID NO:108所示的CDR1，SEQ ID NO:138所示的CDR2，SEQ ID NO:166所示的CDR3；
（31）SEQ ID NO:109所示的CDR1，SEQ ID NO:139所示的CDR2，SEQ ID NO:167所示的CDR3；
（32）SEQ ID NO:110所示的CDR1，SEQ ID NO:140所示的CDR2，SEQ ID NO:168所示的CDR3；
（33）SEQ ID NO:111所示的CDR1，SEQ ID NO:118所示的CDR2，SEQ ID NO:144所示的CDR3；
（34）SEQ ID NO:112所示的CDR1，SEQ ID NO:126所示的CDR2，SEQ ID NO:152所示的CDR3；
（35）SEQ ID NO:113所示的CDR1，SEQ ID NO:141所示的CDR2，SEQ ID NO:170所示的CDR3；
（36）SEQ ID NO:114所示的CDR1，SEQ ID NO:142所示的CDR2，SEQ ID NO:171所示的CDR3；
（37）SEQ ID NO:115所示的CDR1，SEQ ID NO:126所示的CDR2，SEQ ID NO:172所示的CDR3；
（38）SEQ ID NO:116所示的CDR1，SEQ ID NO:143所示的CDR2，SEQ ID NO:174所示的CDR3；
（39）SEQ ID NO:117所示的CDR1，SEQ ID NO:118所示的CDR2，SEQ ID NO:144所示的CDR3。
3.针对甲型肝炎病毒的单域抗体, 其特征在于，所述单域抗体为SEQ ID NO: 1-41中的任意一条所示的抗体链。
4.根据权利要求3所述的针对甲型肝炎病毒的单域抗体, 其特征在于，抗体链的编码序列分别如SEQ ID NO：42-82所示。
5.根据权利要求1-4任意一项所述的针对甲型肝炎病毒的单域抗体, 其特征在于，所述的抗体序列还包括框架区FR；所述框架区FR包括FR1、FR2、FR3和FR4的氨基酸序列；框架区FR的氨基酸序列分别为：
SEQ ID NO:175-195任意一条所示的FR1；
SEQ ID NO:196-222任意一条所示的FR2；
SEQ ID NO:223-252任意一条所示的FR3；
SEQ ID NO:253-254任意一条所示的FR4。
6.根据权利要求1-5任意一项所述的针对甲型肝炎病毒的单域抗体, 其特征在于，所述的单域抗体为VHH。
7.一种核苷酸分子，其特征在于，该核苷酸分子编码权利要求1-5任意一项所述的针对甲型肝炎病毒的单域抗体，其核苷酸序列如SEQ ID NO：42-82中的任意一条所示。
8.一种表达载体，其特征在于，其包含权利要求7所述核苷酸分子。
9.一种宿主细胞，其特征在于，其可以表达出权利要求1-5任意一项所述的针对甲型肝炎病毒的单域抗体，或其包含权利要求8所述的表达载体。
10.权利要求1-5任意一项所述的针对甲型肝炎病毒的单域抗体的Fc融合抗体。
CN201911398827.1A 2019-12-30 2019-12-30 针对甲型肝炎病毒的单域抗体及其衍生蛋白 Active CN111138533B (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201911398827.1A CN111138533B (zh) 2019-12-30 2019-12-30 针对甲型肝炎病毒的单域抗体及其衍生蛋白

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201911398827.1A CN111138533B (zh) 2019-12-30 2019-12-30 针对甲型肝炎病毒的单域抗体及其衍生蛋白

Publications (2)

Publication Number Publication Date
CN111138533A true CN111138533A (zh) 2020-05-12
CN111138533B CN111138533B (zh) 2022-06-10

Family

ID=70522046

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201911398827.1A Active CN111138533B (zh) 2019-12-30 2019-12-30 针对甲型肝炎病毒的单域抗体及其衍生蛋白

Country Status (1)

Country Link
CN (1) CN111138533B (zh)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111718415A (zh) * 2020-07-03 2020-09-29 上海洛启生物医药技术有限公司 一种抗tigit纳米抗体及其应用
CN116514965A (zh) * 2023-06-15 2023-08-01 上海精翰生物科技有限公司 甲型肝炎病毒抗体及其应用
CN116731175A (zh) * 2023-05-19 2023-09-12 四川大学 一种抗cd47的纳米抗体及其制备方法和应用

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040260067A1 (en) * 2001-11-07 2004-12-23 Schofield Darren J. Anti-hepatitis a virus antibodies
WO2011114353A1 (en) * 2010-03-19 2011-09-22 Indian Immunologicals Limited Monovalent human anti-hepatitis a virus antibody and uses thereof
CN103923881A (zh) * 2013-08-09 2014-07-16 北京科兴生物制品有限公司 甲肝病毒单克隆抗体及其应用
CN103923882A (zh) * 2013-08-09 2014-07-16 北京科兴生物制品有限公司 甲肝病毒单克隆抗体及其应用

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040260067A1 (en) * 2001-11-07 2004-12-23 Schofield Darren J. Anti-hepatitis a virus antibodies
WO2011114353A1 (en) * 2010-03-19 2011-09-22 Indian Immunologicals Limited Monovalent human anti-hepatitis a virus antibody and uses thereof
CN103923881A (zh) * 2013-08-09 2014-07-16 北京科兴生物制品有限公司 甲肝病毒单克隆抗体及其应用
CN103923882A (zh) * 2013-08-09 2014-07-16 北京科兴生物制品有限公司 甲肝病毒单克隆抗体及其应用

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
RAN ZHANG等: "Functional recombinant human anti-HAV antibody expressed in milk of transgenic mice", 《TRANSGENIC RESEARCH》 *
马秋平等: "人免疫球蛋白中甲肝抗体效价ELISA检测方法的建立及验证", 《中国生物制品学杂志》 *
黄建锋等: "甲型肝炎病毒单克隆抗体细胞株的建立及鉴定", 《海南医学》 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111718415A (zh) * 2020-07-03 2020-09-29 上海洛启生物医药技术有限公司 一种抗tigit纳米抗体及其应用
CN116731175A (zh) * 2023-05-19 2023-09-12 四川大学 一种抗cd47的纳米抗体及其制备方法和应用
CN116731175B (zh) * 2023-05-19 2024-03-08 四川大学 一种抗cd47的纳米抗体及其制备方法和应用
CN116514965A (zh) * 2023-06-15 2023-08-01 上海精翰生物科技有限公司 甲型肝炎病毒抗体及其应用
CN116514965B (zh) * 2023-06-15 2023-11-10 上海精翰生物科技有限公司 甲型肝炎病毒抗体及其应用

Also Published As

Publication number Publication date
CN111138533B (zh) 2022-06-10

Similar Documents

Publication Publication Date Title
CN111320694B (zh) 一种由重链抗体的可变区组成的纳米抗体gn2及其制备方法和应用
CN110872351B (zh) 一种特异性结合gpc3蛋白的纳米抗体gn1及其制备方法和应用
CN111138533B (zh) 针对甲型肝炎病毒的单域抗体及其衍生蛋白
CN111018985B (zh) 针对牛血清白蛋白bsa的单域抗体的应用
CN111057148B (zh) 针对牛血清白蛋白bsa的单域抗体及其衍生蛋白
CN113150136B (zh) 新型冠状病毒n蛋白单克隆抗体的制备
CN103709251B (zh) 全人源抗cd26抗体及其应用
CN110642948B (zh) B7-h3纳米抗体、其制备方法及用途
CN114262377B (zh) 一种阻断cd70与其配体cd27结合的抗人cd70纳米抗体的制备方法及其编码序列
CN111138532B (zh) 针对甲型肝炎病毒的单域抗体的应用
CN108892723B (zh) 用于检测猪流行性腹泻病毒的单域重链抗体、制备方法及应用
CN103641917B (zh) 抗cd26抗体及其应用
CN112028997A (zh) 抗ceacam5纳米抗体
CN110423277B (zh) Pd-1的纳米抗体及其临床应用
CN109734804B (zh) 针对H3K64Ac/H3K64片段的纳米抗体及其应用
CN113061187B (zh) 一种抗人msln单克隆抗体或其抗原结合片段
CN113683684B (zh) 抗乙肝病毒表面抗原抗体、抗体对、含有其的检测试剂及试剂盒
CN103724431B (zh) 一种人源抗cd26抗体及其应用
CN113912729B (zh) 针对sST2的单域抗体及其衍生蛋白和应用
CN116496395B (zh) 一种结合Dsg3的单克隆抗体及其应用
CN117866100B (zh) 针对d-二聚体的单域抗体或其抗原结合片段及其相关生物材料与应用
CN114605555B (zh) 一种抗新型冠状病毒SARS-CoV-2的双特异中和抗体及其应用
CN116987194B (zh) 人st2抗原的模拟表位肽的抗独特型纳米抗体及应用
US20230365659A1 (en) Sars-cov-2-binding peptide
CN110054675B (zh) 免疫原性多肽以及抗ttc36抗体cp4-3及应用

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant