Cloning and prokaryotic expression of phosphatidylinositol transfer protein alpha gene in Taenia saginata asiatica and the immunologic analysis of the recombinant protein
Abstract:To clone and express the novel phosphatidylinositol transfer protein alpha gene in Taenia saginata asiatica (T.a.PITPα) in order to analyze the immunogenicity of the recombinant protein. By screening the full length cDNA plasmid library,the coding region of T.a.PITP α was amplified with PCR, and cloned into the prokaryotic expression vector pET-28a ( + ) and then expressed in E. col i BL21 with IPTG induction. The recombinant protein was detected by SDS-PAGE and purified by Ni-IDA affinity chromatography. And its immunogenicity was analysed by Western blotting. It was confirmed by PCR, double enzyme digestiom and DNA sequencing that the recombinant expression plasmid was successfully constructed. The expression products were obtained and purified by Ni-IDA affinity chromatography. Western blot analysis of T.a.PITP α recombinant protein testified that the recombinant protein could be recognized by immunized swine serum and could efficiently elicit specific antibodies in SD rats, indicating its immunogenicity. A novel gene coding PITP α of Taenia saginata asiatica was cloned, expressed,purified identification successfully. The purified protein of Ta PITP α will be of importance for further research on the biological function of this gene.
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