| 人肠道病毒71型VP0蛋白的原核表达及多克隆抗体制备 |
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| 引用本文： | 冯嫣芳,刘庆伟,库志强,闻洁君,单虎,黄忠.人肠道病毒71型VP0蛋白的原核表达及多克隆抗体制备[J].细胞与分子免疫学杂志,2011(5):535-538. |
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| 作者姓名： | 冯嫣芳 刘庆伟 库志强 闻洁君 单虎 黄忠 |
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| 作者单位： | 青岛农业大学动物科技学院;中国科学院上海巴斯德研究所分子病毒学与免疫学重点实验室; |
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| 基金项目： | 中国科学院“百人计划”项目(KSCX2-YW-BR-2); 生化工程国家重点实验室开放基金项目(2010KF-07) |
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| 摘 要： | 目的:原核表达并纯化人肠道病毒71型(EV71)VP0蛋白,免疫豚鼠制备多克隆抗体,并鉴定其用于EV71检测的反应性和特异性。方法:PCR方法扩增VP0基因,构建原核表达质粒pET-VP0并转化大肠杆菌,诱导表达并纯化VP0重组蛋白,免疫豚鼠制备抗VP0多克隆抗体,ELISA方法检测抗VP0抗体效价,细胞免疫荧光和Western blot方法鉴定抗体特异性。结果:VP0重组蛋白在大肠杆菌BL21中高效表达,制备的抗VP0抗体效价为1∶106。细胞免疫荧光及Western blot检测结果显示,抗VP0多克隆抗体可以识别原核表达及EV71感染细胞中的VP0蛋白。结论:原核表达了EV71的VP0蛋白并制备出抗VP0多克隆抗体,为EV71的临床诊断、疫苗开发和分子病毒学研究提供了新的研究工具和手段。
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| 关 键 词： | 手足口病 人肠道病毒71型 VP0 多克隆抗体 |
Expression of VP0 protein of enterovirus 71 in Escherichia coli and generation of the corresponding polyclonal antibodies in guinea pigs |
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| FENG Yan-fang,LIU Qing-wei,KU Zhi-qiang,WEN Jie-jun,SHAN Hu,HUANG Zhong College of Animal Science & Technology,Qingdao Agricultural University,Qingdao ,Key Laboratory of Molecular Virology & Immunology,Institute Pasteur of Shanghai,Chinese Academy of Sciences,Shanghai ,China.Expression of VP0 protein of enterovirus 71 in Escherichia coli and generation of the corresponding polyclonal antibodies in guinea pigs[J].Journal of Cellular and Molecular Immunology,2011(5):535-538. |
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| Authors: | FENG Yan-fang LIU Qing-wei KU Zhi-qiang WEN Jie-jun SHAN Hu HUANG Zhong College of Animal Science & Technology Qingdao Agricultural University Qingdao Key Laboratory of Molecular Virology & Immunology Institute Pasteur of Shanghai Chinese Academy of Sciences Shanghai China |
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| Institution: | FENG Yan-fang1,2,LIU Qing-wei2,KU Zhi-qiang2,WEN Jie-jun2,SHAN Hu1,HUANG Zhong2 1College of Animal Science & Technology,Qingdao Agricultural University,Qingdao 266109,2Key Laboratory of Molecular Virology & Immunology,Institute Pasteur of Shanghai,Chinese Academy of Sciences,Shanghai 200025,China |
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| Abstract: | AIM: To obtain recombinant VP0 protein of enterovirus 71,and generate the corresponding VP0-specific polyclonal antibodies,for molecular detection and characterization of EV71.METHODS: The VP0 gene was amplified by PCR and cloned into vector pET26b to make pET-VP0 for the prokaryotic expression of VP0.The recombinant VP0 protein was expressed in E.coli BL21 harboring pET-VP0,purified from inclusion bodies,renatured,and subsequently used to immunize guinea pigs.The resultant antisera were evaluated for anti-... |
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| Keywords: | HFMD enterovirus 71 VP0 polyclonal antibody |
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