Prokaryotic expression and antigenicity analysis of TsP53 gene of Trichinella spiralis

A TsP53 gene fragment was amplified by RT-PCR from the total RNA of Trichinella spiralis muscle larvae and cloned into the expression vector pET30a.It was identified by PCR,restriction enzyme digestion and sequencing,after transformation into Escherichia coli DH5α.The pET30a-TsP53 plasmid was transformed into E.coli BL21(DE3) and induced by IPTG for expression.The expressed product was analyzed by SDS-PAGE and the antigenicity of the purified recombinant protein was identified by indirect ELISA with T.spiralis-infected swine serum.It was demonstrated that after restriction enzyme digestion,PCR amplification and sequencing analysis,the constructed pET30a-TsP53 contained an insert of 1176bp encoding 391 amino acids,which was 99% homologous to T.spiralis P53 gene in GenBank.The recombinant protein was highly expressed in the form of inclusion bodies with the molecular weight of about 50ku.