Cloning,expression and identification of the 14-3-3 gene of Trichomonas vaginalis

Objective To clone and express 14-3-3 gene of Trichomonas vaginalis.Methods Special primers were designed on the basis of the reported T.vaginalis 14-3-3 gene.The 14-3-3 gene was amplified by PCR from the total DNA of T.vaginalis and was cloned into pMD-18-T to construct pMD-14-3-3.The plasmid pMD-14-3-3 was then digested with restriction ribozymes and subcloned into the prokaryotic expression plasmid pGEX-T to construct pGEX-T-Tv-14-3-3.It was then expressed in E.coli BL21(DE3) induced with IPTG.The fusion product was identified by SDS-PAGE and Western blot.Results A prokaryotic expression vector of the 14-3-3 gene was constructed and expressed in Escherichia.coli.Induced with IPTG,the expressed recombinant protein was detected as a band of 27 ku by SDS-PAGE.A special reaction band to anti-14-3-3 sera was observed in Western blot.Conclusion The fusion protein of the 14-3-3 gene was successfully expressed in prokaryotic cells.