Construction of the esat6-mpt64 Fused Eukaryotic Expression Vector of Mycobacterium tuberculosis and its Expression

Objective:To Construction the fused eukaryotic expression vector of Mycobacterium tuberculosis esat6-mpt64 gene.Methods Results:The esat6 and mpt64 genes were amplified by PCR from genome of M.tuberculosis H37Rv strain,and first inserted into cloning vector pGEM-T-easy.After sequencing was conformed,the genes were subcloned to eukaryotic expression vector pCDNA3.1(+).The recombinant plasmid pCDNA-esat6-mpt64 was transfected into COS-7 cells with LipofectAMINE2000.The expression was detected by RT-PCR and indirect immunofluorescence.Conclusion:Eukaryotic recombinant plasmid carrying esat6-mpt64 fused gene was constructed successfully.The esat6-mpt64 fused gene can be expressed in COS-7 cell.