Construction and expression of eukaryotic co-expression plasmid containing Mycobacterium tuberculosis Hsp65 and Ag85A genes

Objective To construct a eukaryotic co-expression plasmid containing Mycobacterium tuberculosis (MTB) Hsp65 and Ag85A genes.Methods Hsp65 and Ag85A genes were obtained by PCR and cloned into eukaryotic expression plasmid pIRES to construct recombinant plasmid pIRES-Hsp65-Ag85A. The recombinant plasmid was transfected into Hela cells and the expression of target genes was detected by RT-PCR.Results Restriction enzyme analysis, PCR and sequencing results showed that the recombinant plasmid pIRES-Hsp65-Ag85A was constructed successfully and the expression of Hsp65 and Ag85A genes could be detected by RT-PCR.Conclusion The eukaryotic expression plasmid pIRES-Hsp65-Ag85A containing Mycobacterium tuberculosis Hsp65 and Ag85A genes was constructed successfully and can co-express Hsp65 and Ag85A in vitro, which lays foundation for further development of DNA vaccine against tuberculosis.