Expression and site directed mutagenesis of serine protease gene of Trichinella

• 1、Institute of Zoonosis, Jilin University

Abstract：Gene T668 of Trichinella spiralis was found which has a high reaction of specific antigen gene by our laboratory, the gene code the Trichinella spiralis serine protease. This experiment used the overlap PCR method of antigen gene of Trichinella spiralis T668 successfully enzyme active center of important sites of site directed mutagenesis transformation, the binding site of the enzyme active center was 259 hydrophilic serine and 240 of the three serine were mutated to hydrophobic alanine., and constructed and purified the soluble expression of the recombinant protein. Western blot analysis showed that the expression of the recombinant pProkaryotic expression and site directedExpression and site directed mutagenesis of T668 gene of Trichinella spiralis mutagenesis of T668 antigen gene of Trichinella spiralisrotein to 26 days positive serum reacted specifically with Trichinella spiralis infection and indicated good reactogenicity. This experiment laid a solid foundation for the study and application of the serine protease in the later stage.

Keywords： Trichinella spiralis Gene T668 site-directed mutagenesis prokaryotic expression