结核分枝杆菌Rv2389基因真核表达质粒的构建及表达

目的:构建结核分枝杆菌Rv2389基因真核表达载体.方法:PCR扩增Rv2389基因,测序正确后克隆入真核表达载体pCDNA3.1(-),重组质粒酶切鉴定正确后以阳离子聚合物转染CHO细胞后, 分别以RT-PCR方法检测mRNA表达和间接免疫荧光技术检测目的蛋白的表达.结果:构建了重组质粒pCDNA- Rv2389, RT-PCR结果证明Rv2389可在CHO细胞中转录,间接免疫荧光检测证明,表达有Rv2389蛋白的细胞着染.结论:成功构建了结核分枝杆菌Rv2389基因的真核表达载体pCDNA-Rv2389,Rv2389基因可以在CHO细胞中表达.

Construction and expression of eukaryotic expression vector of Mycobacterium tuberculosis Rv2389

AIM: To construct the eukaryotic expression vector encoding Mycobacterium tuberculosis Rv2389 and express it in CHO cells. METHODS: The gene encoding Rv2389 protein were amplified by polymerase chain reaction(PCR)from genome of Mycobacterium tuberculosis H37Rv strain. After sequenced, Rv2389 gene segments were subcloned into eukaryotic expression vector pCDNA3.1(-). The recombinant plasmid pCDNA-Rv2389 were transfected into CHO cells with liposome. The expressions of mRNA and protein encoded by this gene were detected respectively with RT-PCR and indirect immunofluoresent technology. RESULTS: Rv2389 was cloned into pCDNA3.1(-) correctly, and its expression at mRNA and protein levels was detected in CHO cells. CONCLUSION: Recombinant eukaryotic plasmid encoding Rv2389 was constructed successfully. The experiment established the basis for further study on the function of Rv2389.