Construction of Prokaryotic Expression Plasmid and Expression in E. coli of AK Gene of Trichomonas Vaginalis

Objective To construct and identify prokaryotic expression plasmid of the hydrogenosomal adenylate kinase of trichomonas vaginalis.Methods The cloning vectors(containing full length of AK DNA and two restriction sites:BamHI and XbaI)were first cut by two restriction enzyme:BamHI and XbaI,and the same as the prokaryotic expression plasmid; then AK DNA and the digested vector were ligated by T4 DNA ligase,and recombinant prokaryotic expression vector was formed.It was certified by PCR and restriction analysis.The compound gene PUC18-AK was expressed in E.coli by IPTG.Results The size of amplified AK gene was 690 bp.The correct recombinant plasmid PUC18-AK was isolated and confirmed by PCR and restriction analysis.The expressed protein AK in E.coli was identified by SDS-PAGE and western blotting.Conclusion The prokaryotic expression plasmid of AK gene was successfully constructed,which was then expressed in E.coli.