| 肠道病毒71型VP1～VP4基因克隆及其表达产物的免疫原性 |
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| 作者姓名： | Song YB Yu N He SJ Chen XX Wang B Che XY Zeng QY |
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| 作者单位： | 1. 南方医科大学珠江医院儿科中心,广东广州,510282
2. 南方医科大学医学检验中心,广东广州,510282 |
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| 基金项目： | 国家重大科技专项课题(2009ZX10004-306); 广州市医药卫生科技重大项目(201102A211007) |
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| 摘 要： | 目的克隆并表达肠道病毒71型VP1~VP4蛋白基因,初步鉴定其免疫原性。方法抽提病毒RNA,经RT-PCR方法分别扩增出VP1~VP4蛋白基因片段,经克隆后,在QIA表达系统中表达,表达产物用8 mol/L尿素洗涤及Ni柱亲和层析纯化后,用肠道病毒71型免疫兔血清和柯萨奇病毒A16型免疫兔血清对重组蛋白进行Western blotting及ELISA鉴定。结果构建的重组质粒pQE30a/VP1~VP4经IPTG诱导,重组蛋白VP1~VP4高效表达并纯化成功,经Western blotting及ELISA证实重组蛋白VP1~VP4可以被免疫兔血清特异识别。结论肠道病毒71型VP1~VP4蛋白表达载体在大肠杆菌M15中高效表达。纯化产物具有较强的免疫原性,为今后EV71亚单位疫苗的研究和检测试剂盒提供了参考。
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| 关 键 词： | 肠道病毒71型 基因克隆 原核表达 免疫原性 |
Gene cloning and immunogenicity analysis of the structural proteins VP1-VP4 of enterovirus 71 |
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| Song YB,Yu N,He SJ,Chen XX,Wang B,Che XY,Zeng QY.Gene cloning and immunogenicity analysis of the structural proteins VP1-VP4 of enterovirus 71[J].Journal of Southern Medical University,2011,31(11):1846-1850. |
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| Authors: | Song Yuan-bin Yu Nan He Si-jie Chen Xin-xin Wang Bin Che Xiao-yan Zeng Qi-yi |
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| Institution: | SONG Yuan-bin1,YU Nan2,HE Si-jie2,CHEN Xin-xin1,WANG Bin1,CHE Xiao-yan2,ZENG Qi-yi1 1Pediatric Center,2Laboratory Testing Center,Zhujiang Hospital,Southern Medical University,Guangzhou 510282,China |
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| Abstract: | Objective To clone the genes encoding the structural proteins VP1-VP4 of enterovirus 71 and investigate the immunogenicity of the expressed recombinant proteins.Methods The VP1-VP4 cDNAs were amplified by RT-PCR from the extracted viral RNA and cloned into pMD19-T vector.The cloned VP1-VP4 genes were then inserted into the multi-cloning sites of plasmid pQE30a and expressed in E.coli M15 with IPTG induction.After washing with 8 mol/L urea and purification with Ni-affinity chromatography,the recombinant prot... |
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| Keywords: | enterovirus 71 gene clone prokaryotic expression immunogenicity |
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