Mycobacterium tuberculosis Rv2389 gene eukaryotic expression plasmid and expression of.doc

 PAGE \* MERGEFORMAT 12 Mycobacterium tuberculosis Rv2389 gene eukaryotic expression plasmid and expression of [Keywords:] Mycobacterium tuberculosis; Rv2389; gene expression Construction and expression of eukaryotic expression vector of Mycobacterium tuberculosis Rv2389 [Abstract] 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 pCDNARv2389 were transfected into CHO cells with liposome. The expressions of mRNA and protein encoded by this gene were detected respectively with RTPCR 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. [Keywords] mycobacterium tuberculosis; Rv2389; gene expression [Abstract] Objective: To construct Mycobacterium tuberculosis Rv2389 gene eukaryotic expression vector. Methods: PCR amplification of Rv2389 gene sequencing, and cloned into the eukaryotic expression vector pCDNA3.1 (-), recombinant plasmid was identified after digestion cation Polymer transfected CHO cells, respectively, mRNA expression was detected by RTPCR and indirect immunofluorescence to detect the target protein expression. Results: The recombinant plasmid pCDNA Rv2389, RTPCR results show that Rv2389 transcription in CHO cells, indirect immunofluorescence assay shows that expression of Rv2389 protein in the cells staining. Conclusion: The successful construction of Mycobacterium tuberculosis Rv2389 gene eukaryotic expression vector pCD