Construction and expression of eukaryotic expression recombinant plasmid of Mycobacterium tuberculosis ESAT-6

Objective To construct eukaryotic expression vector of ESAT-6 and investigate transient expression of its protein in COS-7 cells.Methods ESAT-6 coding sequence esxA was amplified from Mycobacterium tuberculosis H37Rv genomic DNA by using PCR.PCR product was cloned into eukaryotic expression vector pcDNA3.1/Zeo (+).Eukaryotic expression vector pcDNA-ESAT-6 were transfected into eukaryotic cells COS-7 by means of DEAE-dextran,and transient expression of its protein was investigated in eukaryotic cells with RT-PCR.Results ESAT-6 coding sequence was successfully inserted into the eukaryotic expression vector pcDNA3.1/Zeo (+) and efficiently expressed in eukaryotic cells (COS-7).Conclusion The recombinant pcDNA-ESAT-6 could be successfully constructed and expressed efficiently in COS-7.