Construction and expression of eukaryotic expression vector of Mycobacterium tuberculosis Rv2389

    XUE Ying, BAI Yinª²Lan, GAO Hui, WANG Liª²Mei, XU Zhiª²Kai

    Department of Microbiology, School of Basic Medicine, Fourth Military Medical University, Xi¬ðan 710033, China

    ¡¾Abstract¡¿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 pCDNAª²Rv2389 were transfected into CHO cells with liposome. The expressions of mRNA and protein encoded by this gene were detected respectively with RTª²PCR 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. ......