| 结核杆菌热休克蛋白70基因的克隆与原核表达 |
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| 引用本文： | 叶菁,隋延仿,陈广生,张秀敏.结核杆菌热休克蛋白70基因的克隆与原核表达[J].细胞与分子免疫学杂志,2003,19(5):443-445. |
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| 作者姓名： | 叶菁 隋延仿 陈广生 张秀敏 |
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| 作者单位： | 第四军医大学基础部病理学教研室,陕西,西安,710032 |
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| 基金项目： | 全军医药卫生科研基金重点项目资助 (No.0 1Z0 84) |
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| 摘 要： | 目的：克隆结核杆菌热休克蛋白70(TBhsp70)基因，并在大肠杆菌中表达。方法：利用PCR技术从结核杆菌H37Rv中扩增Hsp70基因，并将其克隆到pUC19中，进行测序。将得到的Hsp70基因克隆到表达载体pGEX-4T-1中，构建重组表达质粒pGEX—TBhsp70，在大肠杆菌DH5α中进行表达。结果：成功地克隆了TBhsp70基因。DNA测序证实，与GenBank公布的序列一致。含pGEX-TBhsp70基因表达质粒的大肠杆菌经IPTG诱导后，能够表达相对分子质量(MR)约为96000的融合蛋白。结论：获得了TBhsp70基因，成功地构建了原核表达质粒pGEX-TBhsp70，并在大肠杆菌得到表达，为其相关研究奠定了一定的基础。
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| 关 键 词： | 结核杆菌 热休克蛋白 克隆 原核表达 |
| 文章编号： | 1007-8738(2003)05-443-03 |
Cloning and prokaryotic expression of heat shock protein 70 gene of Mycobac-terium tuberculosis |
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| Jing Ye,Yan-fang Sui,Guang-sheng Chen,Xiu-min Zhang.Cloning and prokaryotic expression of heat shock protein 70 gene of Mycobac-terium tuberculosis[J].Journal of Cellular and Molecular Immunology,2003,19(5):443-445. |
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| Authors: | Jing Ye Yan-fang Sui Guang-sheng Chen Xiu-min Zhang |
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| Institution: | Department of Pathology, Fourth Military Medical University, Xi'an 710032, China. |
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| Abstract: | AIM: To clone the heat shock protein 70 (Hsp70) gene of Mycobacterium tuberculosis and express it in E.coli . METHODS: Hsp70 gene was amplified by PCR from Mycobacterium tuberculosis H37Rv, then inserted into pUC19 vector and sequenced. The recombinant expression plasmid pGEX TBhsp70 was constructed and expressed in E.coli . RESULTS: Hsp70 gene was obtained form Mycobacterium tuberculosis and its sequence was identical with that reported in GenBank. The E.coli . containing pGEX TBhsp70 could express a protein with M r being 96 000 after induction of IPTG. CONCLUSION: A confirmed Hsp70 gene is cloned and expressed in E.coli successfully, which lay the foundation for the related research of the gene. |
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| Keywords: | Mycobacterium tuberculosis heat shock proteins cloning prokaryotic expression |
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