| 空肠弯曲菌CadF蛋白的原核表达与产物分析 |
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| 引用本文： | 周益装,张茂俊,肖迪,孟凡亮,张建中. 空肠弯曲菌CadF蛋白的原核表达与产物分析[J]. 中国病原生物学杂志, 2010, 0(5) |
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| 作者姓名： | 周益装 张茂俊 肖迪 孟凡亮 张建中 |
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| 作者单位： | 中国疾病预防控制中心传染病预防控制所; |
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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(No.2008ZX10004-002); 中国疾病预防控制中心优秀青年基金项目(No.2009A102) |
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| 摘 要： | 目的构建空肠弯曲菌cadF基因重组表达质粒pET30a(+)-cadF,分析其在大肠埃希菌中的表达情况及其融合蛋白的抗原性。方法用PCR方法扩增cadF基因,构建重组克隆质粒和表达质粒,经菌落PCR、双酶切和测序鉴定后在E.coli(DE3)原核表达系统中诱导表达,SDS-PAGE和飞行时间质谱鉴定表达蛋白,Western blot分析其抗原性。结果含重组表达载体pET-30a(+)-cadF中的cadF基因序列经测序证实与出发菌株cadF基因序列100%同源,并成功诱导表达CadF蛋白,Westernblot显示该CadF融合蛋白能被抗空肠弯曲菌多克隆兔血清识别。结论成功构建了空肠弯曲菌cadF基因重组表达载体,表达产物具有抗原性。
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| 关 键 词： | 空肠弯曲菌 CadF 原核表达 抗原性 |
Prokaryotic expression and determination of protein CadF from Campylobacter jejuni |
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| ZHOU Yi-zhuang,ZHANG Mao-jun,XIAO Di,MEN Fan-liang,ZHANG Jian-zhong. Prokaryotic expression and determination of protein CadF from Campylobacter jejuni[J]. Journal of Pathogen Biology, 2010, 0(5) |
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| Authors: | ZHOU Yi-zhuang ZHANG Mao-jun XIAO Di MEN Fan-liang ZHANG Jian-zhong |
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| Affiliation: | ZHOU Yi-zhuang,ZHANG Mao-jun,XIAO Di,MEN Fan-liang,ZHANG Jian-zhong (National Institute for Communicable Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China) |
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| Abstract: | Objective To construct a recombinant plasmid pET30a(+)-cadF of Campylobacter jejuni,investigate its expression in E. coli,and determine the antigenicity of fusion proteins. Methods The cadF gene was amplified by PCR and then a recombinant clone and expression plasmid were constructed. After identification using PCR,double restriction endonuclease digestion,and DNA sequencing analysis,the recombinant was transformed into E. coli (DE3). A recombinant CadF protein (rCadF) was induced,expressed in an E. coli (D... |
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| Keywords: | Campylobacter jejuni CadF prokaryotic expression antigenicity |
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