Construction and Expression of Multi-epitopes Gene of Treponema pallidum and Immunoresponse of Recombinant Protein / 中华医院感染学杂志

ABSTRACT

OBJECTIVE To develop the strong and the specific multi-epitope antigen for the exploiting diagnosis of Treponema pallidum.METHODS The immuno-dominant epitopes of Tp0453 and Tp17 were amplified by PCR respectively,and subcloned into the expression vector pQE32 to generate multi-epitopes recombinant plasmid pQE32/Tp0453-17.The recombinant protein was expressed in Escherichia coli M15.The immunoresponse of recombinant fusion protein was analyzed by Western blot.RESULTS The multi-epitopes recombinant plasmid was successfully constructed,enzyme digestion analysis and sequencing showed that the inserted target genes were Tp0453 and Tp17 gene,compared with the gene reported by GenBank,it had 100% similarity;SDS-PAGE analysis showed the recombinant plasmid could be expressed in M15,its relatively molecular mass(Mr) of expressed product was about 52.0?103.The Western blot result showed the recombinant protein could be recognized by anti-T.pallidum positive serum.CONCLUSIONS The expressed multi-epitopes recombinant antigen showed excellent immunoresponse.The results lay the foundation for research on development of quick diagnostic kit applying to detection of T.pallidum infection.