Prokaryotic clone, expression and immunological analysis of recombinant Schistosoma japonicum cyclophilin B / 国际检验医学杂志

ABSTRACT

Objective To clone and express Schistosoma j aponicum cyclophilin B (SjCyPB) gene in E.coli,and to identify and analyze the immunity of recombinant proteins.Methods A pair of specific primers was designed according to GenBank of Schistosoma japonicum sequence.SjCyPB gene was amplify by PCR and then connected to pET28 vector.The recombinant plasmid pET28a (+)-SjCyPB was constructed and transformed into E.coli BL21 cell line,the recombinant plasmid was identified by double enzyme digestion and sequence analysis.After induced by isopropyl-B-D-thiogalaetoside (IPTG),the expressed recombinant proteinwas purified by affinity-chromatography,and then verified by Western blotting.Rats were immunized recombinant SjCyPB,andthe SjCyPB-specific IgG was detected by ELISA.Results SjCyPB gene was successfully inserted into pET28a(-) vecter which identified by double enzyme digestion and sequence analysis.Recombinant SjCyPB protein was highly expressed in E.coli.The Western Blotting analysis confirmed that the recombinant protein could specifically combine to S.japonicum-infected rabbit serum.Using recombinant protein to immunize rats,the SjCyPB-specific IgG antibody titer was 1 ∶ 51 200 detected by ELISA.Conclusion The recombinant SjCyPB is successfully constructed,and recombinant SjCyPB has immunogenicity and antigenicity.