Cloning and prokaryotic expression of TvRab11C gene of Trichomonas vaginalis

Objective To clone the TvRab11C gene of Trichomonas vaginalis and express in prokaryotic cells.Methods The TvRab11C gene was amplified by PCR from T.vaginalis and inserted into prokaryotic expression vector pET-28a.The constructed recombinant plasmid pET-28a-TvRab11C was transformed to E.coli BL21(DE3) and induced by IPTG.The expressed product was analyzed for solubility by SDS-PAGE and for reactogenicity by Western blot.Results Both restriction analysis and sequencing proved that recombinant plasmid pET-28a-TvRab11C was constructed correctly.The expressed recombinant protein,with a relative molecular mass of about 30 000,mainly existed in a form of inclusion body,and was recognized by polyclonal antibody against T.vaginalis.Conclusion The TvRas gene of T.vaginalis was successfully cloned and expressed in E.coli BL21(DE3),which laid a foundation of further study on relationship of TvRas gene and protein to the parasitic ability and pathogenicity of T.vaginalis.