Cloning , expression and immunogenicity analysis of Ta 8 gene of Taenia asiatica oncospheres

The novel gene named oncosphere 8kDa gene of Taenia asiatica(Ta8)was cloned and expressed in order to evaluate the value of the recombinant protein in the diagnosis.The full-length cDNA of Ta8gene was cloned fromTaenia asiatica by using RT-PCR and cloned into the prokaryotic expression vector pET-28a(+),and then expressed in E.coli BL21 with IPTG induction.The recombinant protein was detected by SDS-PAGE and purified by Ni-IDA affinity chromatography.Its immunogenicity was examined by Western blot.PCR,double enzyme digestion and DNA sequencing confirmed that the recombinant expression plasmid was successfully constructed.The expression products were obtained and purified by Ni-IDA affinity chromatography.Western blot analysis of Ta8recombinant protein testified that it could be recognized by immunized SD rat serum while could not be recognized by normal SD rat,therefore indicated its immunogenicity.A novel gene coding Ta8of Taenia asiatica was cloned,expressed,purified and identified successfully,which proofing that Ta8protein had immunogenicity.Additionally,the Ta8gene was also detected in scolex and mature and gravid proglottids of adult Taenia asiatica.