Construction,expression and identification of eukaryotic expression vector carrying Schistosoma japonicum gene coding lysophospholipase

Abstract

：Objective Schistasomajaponicum(S.japonicum)lysophospholipase gene(Sjl539)from cDNA of S japonicum adult wormswas amplified and subcloned into eukaryotic expression vector pcDNA3.1(+)for expression ofrecombinant antigen and immunogenicity analysis.Methods Total RNA of S.japonicum wasextracted to generato cDNA by RT-PCR.The Sj1539 gent was amplified.The DNA fragment wassubcloned into eukaryofic expression vector pcDNA3.1(+)following insertion andamplification in pGEM-T.The recombinant plasmid was transfected into human cervicalcarcinoma cell strain(Hela cells)and expression products were identified by Westernblotting.Results The size of PCR product was approximately 684 bp.It was confirmed thatSj1539 gene had been inserted successfully by the recombinant plasmid digested with twoenzymes and PCR.It was verified that the expression product could react withS.japonicum-infected rabbit serum by Western blotting and the molecular weight wasapproximately 25×103.Conclusions The eukaryotie expression vector carrying Sj1539 genehas been established and the expression product has been obtained.