| 卡介苗热休克蛋白16.3基因的克隆与原核表达 |
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| 引用本文： | 胡松,范雄林,姚炜,王芳,黄汉菊. 卡介苗热休克蛋白16.3基因的克隆与原核表达[J]. 华中医学杂志, 2006, 30(6): 495-496 |
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| 作者姓名： | 胡松 范雄林 姚炜 王芳 黄汉菊 |
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| 作者单位： | 江汉大学医学与生命科学学院病原学教研室;430030,武汉,华中科技大学同济医学院基础医学院 |
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| 摘 要： | 目的 克隆卡介苗(BCG)的热休克蛋白16.3基因(hsp16.3),并在大肠杆菌中表达.方法 利用PCR技术从BCG中扩增hsp16.3基因,并将其克隆到质粒pProEX HTb中,进行测序.将得到的重组表达质粒pProEX HTb-hsp16.3在大肠杆菌BL21中进行表达.结果 成功地克隆了BCG hsp16.3基因.经DNA测序证实,与Gen Bank公布的序列一致.含pProEX HTb- hsp16.3重组质粒的大肠杆菌BL21经诱导后,能够表达相对分子质量约为16KDa的融合蛋白.结论 获得了BCG hsp16.3基因,成功构建原核表达质粒pProEX HTb- hsp16.3,并在大肠杆菌得到高效表达.
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| 关 键 词： | 卡介苗 热休克蛋白类 克隆载体 |
| 修稿时间： | 2006-08-17 |
Cloning and prokaryotic expression of heat shock protein 16.3 gene of Mycobacterium bovis BCG |
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| Hu song,Fan Xionglin,Yao wei et al.TongJi Medical College of HuaZhong University of Science and Technology,Wuhan. Cloning and prokaryotic expression of heat shock protein 16.3 gene of Mycobacterium bovis BCG[J]. Central China Medical Journal, 2006, 30(6): 495-496 |
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| Authors: | Hu song Fan Xionglin Yao wei et al.TongJi Medical College of HuaZhong University of Science Technology Wuhan |
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| Affiliation: | Hu song,Fan Xionglin,Yao wei et al.TongJi Medical College of HuaZhong University of Science and Technology,Wuhan 430030 |
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| Abstract: | Objective To clone the heat shock protein 16.3(hsp16.3)gene of BCG and express it in E.coli.Methods hsp16.3 gene was amplified by PCR from BCG genome DNA,then inserted into pProEX HTb vector and sequenced.The recombinant expression plasmid pProEX HTb-hsp16.3 was constructed and expressed in E.coli BL21 strain. Results hsp16.3 gene was obtained form BCG and its sequence was identical with that reported in Gen Bank.The E.coli BL21 strains containing pProEX HTb-hsp16.3 expressed a 16 KDa protein after induction of IPTG.Conclusion A confirmed BCG hsp16.3 gene was cloned and expressed in E.coli successfully,which lays the foundation for structural and functional research of HSP16.3 protein. |
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| Keywords: | BCG Heat shock protein Genetic rectors |
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