Cloning and prokaryotic expression of WD40 gene of Taenia saginata asiatica and immunogenicity of recombinant protein

Objective To clone and express the novelgene named WD40 of Taenia saginata asiatica in order to analyze the immunogenicity of the recombinant protein.Methods By scr eening the full length cDNA plasmid library,the coding region of WD40 was amplified with PCR,and cloned into the prokar yotic ex pression vector pET-28a(+)and then expressed in E.coli BL21 with IPTGinduction.The recombinant protein was detected by SDS-PAGE and purified by NiIDA affinity chromatography,and then its immunogenicity was analyzed by Western Blotting.Results PCR,double enzyme digestio n and DNA sequencing confirmed that the recombinant expression plasmid was successfully constructed.The expression products were obtained and purified by Ni-IDA affinity chromato graphy.Western Blot analysis of WD40 recombinant protein test ified that the reco mbinant protein could be recognized by the serum of immunized swine and the patients.Conclusion A novel gene coding WD40 of Taenia saginata asiatica was cloned,expressed,purified successfully.The purified protein of WD40 will be of importance for further research on the biological function of the gene.