Expression and Purification of ESAT-6 Protein of Mycobacterium Tuberculosis

To construct the recombinant prokaryotic expression vector containing the ESAT-6 gene of Mycobacterium bovis,purification,expression the fusion protein identified.Methods:We cloned the ESAT-6 gene from mycobacterium bovis chromosomal DNA by using PCR,into the expression vector pET-22b(+) with the NdeI,EcorI two enzymesites,identified the recombinant vector by PCR,sequencing,the conatructed recombinant plasmid was transformed into E.coli BL21(DE3) polys and the recombinant proteins were expressed after induction with IPTG;The target proteins were identified by Western-blot and purified by using affinity chromatograghy.Results:The sequencing result of ESAT-6 gene was the same as the Genbank.6kDa protein was found by SDS-PAGE and Western-blot.Pure target protein has been obtained with the protocol we developed.Conclusion:The ESAT-6 fusion protein was successfully expressed and purified,identified.