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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 96 0 0 0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。
Objective: To clone Mycobacterium tuberculosis heat shock protein 70 (TBhsp70) gene and express in E. coli. Methods: Hsp70 gene was amplified from Mycobacterium tuberculosis H37Rv by PCR and cloned into pUC19 for sequencing. The resulting Hsp70 gene was cloned into the expression vector pGEX 4T 1 to construct the recombinant expression plasmid pGEX TBhsp70 and expressed in E. coli DH5α. Results: The TBhsp70 gene was successfully cloned. DNA sequencing confirmed that the sequence published in GenBank consistent. After induced by IPTG, E.coli harboring pGEX TBhsp70 gene expression plasmid can express a fusion protein with molecular weight (Mr) of about 9600. CONCLUSION: The TBhsp70 gene was obtained and the prokaryotic expression plasmid pGEX TBhsp70 was successfully constructed and expressed in E. coli, which laid the foundation for its related research.