结核分枝杆菌Rv1884基因真核表达质粒的构建及表达

目的:构建结核分枝杆菌Rv1884基因真核表达载体。方法:PCR扩增Rv1884基因,测序正确后克隆入真核表达载体pcDNA3.1(-);经酶切鉴定正确的重组质粒酶以阳离子聚合物转染P815细胞后,以RT-PCR方法检测mRNA的表达,以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-Rv1884;RT-PCR结果证明Rv1884可在P815细胞中转录;用间接免疫荧光检测,表达有Rv1884蛋白的细胞着染。结论:构建了结核分枝杆菌Rv1884基因的真核表达载体pcDNA-Rv1884,Rv1884基因可以在P815细胞中表达。

Construction of the Fused Eukaryotic Expression Vector of Mycobacterium tuberculosis Rv1884 Gene and its Expression

Objective:To construct eukaryotic expression vector encoding Mycobacterium tuberculosis Rv1884 gene.Methods:The gene encoding Rv1884 protein were amplified by PCR from genome of M.tuberculosis H37Rv strain.After sequenced,Rv1884 gene segments were subcloned into eukaryotic expression vector pcDNA3.1(-).The recombinant plasmid pcDNA-Rv1884 were transfected into P815 cells with liposome.The expressions of mRNA and the expressed of proteins encoded by this gene were detected with RT-PCR and indirect immuno fluensent respectively.Results:Rv1884 gene was cloned into pcDNA3.1(-)correctly,and its expression was detected in P815 cells.Conclusion:Eukaryotic recombinant plasmids encoding Rv1884 were constructed successfully.The results established the basis for further study of the function of Rv1884.