Cloning, expression and sequence analysis of cysteine-rich secretion protein 2 gene from Taenia saginata asiatica

ObjectiveTo search and identify novel gene by screen ing the cDNA library of adult Taenia saginata asiatica, and to clone and express the gene so as to lay a foundation for the further study of Ta CRISP gene's function.MethodsBy the bioinformatics analyzing tools in bioinformatics webs site,a Ta CRISP 2 full-length gene from the Ta eniasa ginata asiatica full-length cDNA plasmid libratory was identified and the coding region sequence and the characteristics of the deduced prote in were analyzed.The coding region of Ta CRISP was amplified with PCR,endonuc lease digestion and cloned in to the prokaryotic expression vector pET-30a(+)and then expressed in E.coli BL 21 with IPTG induction.The recombinant protein was detected by SDS-PAGE.ResultsA novel cDNA sequence encoding Ta CRISP with a molecular mass of 269738,pI 5.96,which was 1090 bp and encoding 239 aa,and with 1aa to 16aa signal peptide,was found from the cDNA library of Taenia saginata asiatica.The recom binant plasmid pET-30a(+)-TaCRISP was constructed and expressed in fusion prote in.ConclusionA novel gene encoding TaCRISP of Taenia saginata asiatica was identified and its prokaryotic expression vectors was expressed in fusion protein successfully.