| 大肠杆菌外膜蛋白OmpW的克隆及在外膜上高表达 |
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| 引用本文： | 邹海杰,吴贤斌,潘建义,赵辅昆.大肠杆菌外膜蛋白OmpW的克隆及在外膜上高表达[J].中国生物工程杂志,2011,31(12):46-50. |
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| 作者姓名： | 邹海杰 吴贤斌 潘建义 赵辅昆 |
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| 作者单位： | 浙江理工大学生命科学学院 杭州 310018 |
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| 基金项目： | 浙江省自然科学基金(Y2090396); 浙江省公益技术研究社会发展项目(2011C23067)资助项目 |
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| 摘 要： | 目的: 利用表达载体pLLP-OmpA实现大肠杆菌K12外膜蛋白OmpW在外膜上高表达。方法: PCR扩增ompW基因,构建重组表达载体pLLP-OmpA-ompW,然后转化大肠杆菌K12,得到在外膜上高表达的菌株。提取该菌外膜蛋白,利用免疫小鼠制备得到的抗血清进行Western blot分析验证高表达的OmpW是否定位于外膜。结果: 成功构建了重组表达载体,经转化后成功筛选到高表达菌株,并经Western blot证实高表达的OmpW定位在外膜上。结论: 首次成功获得OmpW在外膜上的高表达,该高表达菌株可为深入研究OmpW在细菌致病机制中的作用及其它功能提供研究基础。
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| 关 键 词： | 大肠杆菌K12 OmpW 重组载体 抗血清制备 |
| 收稿时间： | 2011-08-03 |
| 修稿时间： | 2011-09-04 |
Cloning and Expression of ompW Location in Outer Membrane of Escherichia coli |
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| ZOU Hai-jie,WU Xian-bin,PAN Jian-yi,ZHAO Fu-kun.Cloning and Expression of ompW Location in Outer Membrane of Escherichia coli[J].China Biotechnology,2011,31(12):46-50. |
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| Authors: | ZOU Hai-jie WU Xian-bin PAN Jian-yi ZHAO Fu-kun |
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| Institution: | ZOU Hai-jie WU Xian-bin PAN Jian-yi ZHAO Fu-kun(School of Life Sciences,Zhejiang Sci-Tech University,Hangzhou 310018,China) |
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| Abstract: | Objective:To achieve the mutant stains of Escherichia coli K12 which up-expressed OmpW located in outer membrane by vector pLLP-OmpA.Methods:The complete sequence of ompW from E.coli K12 was cloned by PCR.The cloned DNA fragments were linked with the plasmid to construct the vector pLLP-OmpA-ompW.The recombinant vector was then transferred into E.coli K12 to obtain the engineering strain.Western bolt analysis with primary antibody against OmpW was used to verify whether the highly expressed OmpW of the stra... |
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| Keywords: | Escherichia coli K12 OmpW Recombination plasmid Antiserum preparation |
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