Fusion Expression of the ESAT-6 Gene of the Tuberculosis Bacterium

Objective To construct the recombinant prokaryotic expression vector containing the Early secretary antigenic target(ESAT-6) gene of Mycobacterium tuberculosis,expression the fusion protein identified.Methods We cloned the ESAT-6 gene from mycoba-cterium tuberculosis by PCR,into the expression Vector pET-32a(+)with the BamH Ⅰ,Hind Ⅲ two enzyme sites,identified the recombinant vector by PCR,sequencing.expressed the protein in E coil.BL21,identified the fusion protein with Western-blotting.Results The sequencing result of ESAT-6 gene was the same as the Genbank.The relative molecular mass of target protein is about 2.5×10~4,has the activity of banding antibody.Conclusion The ESAT-6 fusion protein was successfully expressed and purified,identified.