| 霍乱弧菌外膜蛋白W的原核表达及抗原性分析 |
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| 引用本文： | 李建华,王忠田,何君,左庭婷,韩雪莲,李岩伟,朱虹,端青. 霍乱弧菌外膜蛋白W的原核表达及抗原性分析[J]. 军事医学科学院院刊, 2011, 35(10): 746-748,753 |
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| 作者姓名： | 李建华 王忠田 何君 左庭婷 韩雪莲 李岩伟 朱虹 端青 |
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| 作者单位： | 1. 军事医学科学院微生物流行病研究所,病原微生物生物安全国家重点实验室,北京,100071
2. 中国兽医药品监察所,北京,100081 |
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| 基金项目： | “艾滋病和病毒性肝炎等重大传染病防治”科技重大专项(2008ZX10004-003) |
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| 摘 要： | 目的在大肠杆菌中表达霍乱弧菌种特异性外膜蛋白（Omp）W,纯化后制备检测用OmpW抗体。方法采用PCR法以霍乱弧菌基因组为模板扩增ompW基因,插入表达载体pET-32a（＋）的多克隆位点,构建重组表达质粒pET-32a-ompW;重组质粒转化至大肠杆菌BL21（DE3）中,筛选阳性重组菌株,经IPTG诱导获得目的蛋白,纯化后免疫新西兰大耳白兔,制备OmpW的多克隆抗体并进行鉴定。结果扩增得到ompW基因片段,并成功构建pET-32a-ompW原核表达系统,重组蛋白OmpW以包涵体形式高效表达;制备的OmpW抗体能与天然的O1群和O139群霍乱弧菌结合。结论霍乱弧菌OmpW可由原核系统高效表达,免疫获得的抗体具有较好的效价,为进一步制备霍乱弧菌检测用抗体奠定基础。
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| 关 键 词： | 外膜蛋白W 霍乱弧菌 原核表达 抗原性 |
Expression of Vibrio cholerae outer membrane protein W and identification of its antigenicity |
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| LI Jian-hua,WANG Zhong-tian,HE Jun,ZUO Ting-ting,HAN Xue-lian,LI Yan-wei,ZHU Hong,DUAN Qing. Expression of Vibrio cholerae outer membrane protein W and identification of its antigenicity[J]. Bulletin of the Academy of Military Medical Sciences, 2011, 35(10): 746-748,753 |
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| Authors: | LI Jian-hua WANG Zhong-tian HE Jun ZUO Ting-ting HAN Xue-lian LI Yan-wei ZHU Hong DUAN Qing |
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| Affiliation: | LI Jian-hua1,WANG Zhong-tian2,HE Jun1,ZUO Ting-ting1,HAN Xue-lian1,LI Yan-wei1,ZHU Hong1,DUAN Qing1* (1.State Key Laboratory of Pathogen and Biosecurity,Institute of Microbiology and Epidemiology,Academy of Military Medical Sciences,Beijing 100071,China,2.China Institute of Veterinary Drug Control,Beijing 100081,China) |
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| Abstract: | ObjectiveTo obtain Vibrio cholerae outer membrane protein W expressed in E.coli(DE3) in order to produce its polyclonal antibody.MethodsThe ompW gene amplified from V.cholerae by PCR was inserted into expression plasmid pET-32a(+) to construct recombinant plasmid pET-32a(+)-ompW that was transformed into E.coli BL21(DE3) with IPTG induction for the expression OmpW.The protein OmpW was purified with degeneration and renaturation and used to immunize rabbits to prepare anti-OmpW antibody,the property of which... |
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| Keywords: | outer membrane protein W Vibrio cholerae prokaryotic expression antigenicity |
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