Cloning and Expression of Schistosoma Japonicum Truncated SJCHGC02377 Protein

Objective To clone and express Schistosoma japonicum SJCHGC02377 protein.Methods The gene encoding Schistosoma japonicum SJCHGC02377 protein was amplified by RT-PCR from Schistosoma japonicum worm total RNA,and was cloned into pMD18-T vector.Positive clones were identified by colony PCR and sequencing with ABI PRISMTM 377XL DNA Sequencer;The insert digested with BamHⅠand SalⅠwas subcloned into prokaryotic expression vector pET-23a(+) which were also digested with BamHⅠand SalⅠ;The recombinant plasmids were transformed into E.coli BL21 and were identified by colony PCR and double enzyme digestion;The recombinant clone was induced with IPTG to express target protein and characterized by SDS-PAGE;Expressed recombinant protein was purified by affinity chromatography.Results The gene fragment encoding Schistosoma japonicum truncated SJCHGC02377 protein was amplified by RT-PCR from Schistosoma japonicum worm total RNA;The insert of positive clone was subcloned into pET-23a(+) correctly;Recombinant protein was expressed in the positive recombinant clone when induced with IPTG and was purified by affinity chromatography from E.coil BL21 lysate.Conclusion The recombinant plasmid expressing Schistosoma japonicum truncated SJCHGC02377 protein was successfully constructed and the recombinant protein was expressed in E.coil BL21.