Cloning and prokaryotic expression of 26kDa glutathione S-transferase gene and recombinat protein immunogenicity analysis of Taenia saginata asiatica

ObjectiveTo clone and express the novel gene named glutathione S-transferase gene(GST)of Taenia saginata asiatica in order to analyze the immunogenicity of the recombinant protein and further research on the biological function of the gene.MethodsBy screening the full length cDNA plasmid library,the coding region of GST was amplified with PCR, and cloned into the prokaryotic expression vector pET-30a(+)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.Analyze its immunogenicity by Western Blotting.ResultsPCR,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 GST recombinant protein testified that the recombinant protein could be recognized by the serum of swine and patient immunized.GST recombinant protein also could elicit efficiently specific antibodies in SD rats,therefore indicated its immunogenicity.ConclusionA novel gene coding GST of Taenia saginata asiatica was cloned, expressed,purified successfully.The purified protein of GST will be of importance for further research on the biological function of the gene.