Cloning and Expression of Ag85C Gene from Mycobacterium Tuberculosis Edman Strain

Objective To clone and express the Ag85C gene of mycobacterium tuberculosis in Escherichia coli,and to study the immunoprotect function to tuberculosis.Methods The mycobacterium tuberculosis of Edman strain of Ag85C gene was amplified through a couple of specific primer by PCR.The reading frame of Ag85C was isolated and cloned into the prokaryotic expression vector pQE30.The recombinant plasmid was transformed into E.coli BL21 after identifying by restriction enzyme digestion,PCR and DNA sequence analysis.The expression product was identified by SDS-PAGE and Western blot after inducing by 0.1mmol/L IPTG.Results The recombinant plasmid of pQE30-Ag85C was constructed successfully and highly expressed a specific 30kDa fusion protein.Conclusions The successfully constructed expression plasmid and the high expression in E.coli will be benefit to the gene's function study and the trituration of new generation vaccine.