结核分枝杆菌Rv2450基因真核表达质粒的构建及表达

Construction of the Fused Eukaryotic Expression Vector of Mycobacterium Tuberculosis Rv2450 and Its Expression

构建结核分枝杆菌Rv2450基因真核表达载体。PCR扩增Rv2450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P815细胞,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-Rv2450,RT-PCR结果证明Rv2450可在P815细胞中转录,用间接免疫荧光检测,表达有Rv2450蛋白的细胞着染。成功构建了结核分枝杆菌Rv2450基因的真核表达载体pCDNA-Rv2450,Rv2450基因可以在P815细胞中表达。

To construct eukaryotic expression vector encoding Mycobacterium tuberculosis Rv2450, the gene encoding Rv2450 protein was amplified by polymerase chain reaction（PCR） from genome of Mycobacterium tuberculosis H37Rv strain. After sequencing the gene was then subcloned into eukaryotic expression vector pCDNA3.1 （ - ）. The recombinant plasmid pCDNA-Rv2450 was transfected into I）815 cells with liposome. The expression was detected with RT-PCR and indirect immuno fluensent. Rv2450 was cloned into pCDNA3.1 （ - ） correctly, and its expression was detected in I）815 cells. Eukaryotic recombinant plasmids encoding Rv2450 were constructed successfully. The results established the basis for further study of the function of Rv2450.