| 引用本文： |
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杨贵珍,刘钟滨,袁建平,胡宝瑜,时晓东,郭晓奎.幽门螺杆菌VacA毒性亚单位在大肠杆菌中的分别表达[J].同济大学学报（医学版）,2004,(1):7-10. [点击复制]
- .Separate Expressions of Toxic Subunits of VacA of Helicoacter Pylori in E.coli[J].Journal of Tongji University(Medical Science),2004,(1):7-10. [点击复制]
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| 摘要: |
| 目的用基因工程方法获取幽门螺杆菌vacA基因毒性相关片段p37和p58,以深入研究vacA的致病机制。方法用PCR技术,扩增幽门螺杆茵vacA基因的两个片段p37、p58,并克隆入原核表达载体pQE-31,构建重组质粒pQE31—p37和pQE31-p58,转化大肠埃希茵M15,IPTG诱导蛋白质表达,用Ni-NTA柱纯化融合蛋白。结果SDS-PAGE分析显示,经IPTG诱导的E.coli如M15表达出相对分子量分别为37KDa和58kda的P37和P58融合蛋白质,SDS-PAGE分析表明,表达的蛋白质经含Ni-NIA的层析柱纯化后为单一蛋白质。结论成功构建了原核表达栽体pQE31-p37和pQE31-p58,获得了高纯度的单一蛋白质,为进一步探讨VacA的生物学活性及VacA抗体的制备鉴定了基础。 |
| 关键词: 幽门螺杆菌,VacA,亚单位,表达 |
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| 修订日期：2003-09-19 |
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| 基金项目:国家教委留学归国基金(2001498) |
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| Separate Expressions of Toxic Subunits of VacA of Helicoacter Pylori in E.coli |
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| Abstract: |
| Objective To gain the active subunits of VacA of Helicobater pylori by gene engineering technique for further studing. Methods Two gene fragments of H. pylori vacA amplified by PCR were separately cloned into prokaryotic expression vector pQE31.The E. col |
| Key words: helicobater pylori,VacA,subunits,expression |
