Prokaryotic Expression of an Antigen Gene of Trichinella spiralis and Identification of the Recombinant Protein / 中国寄生虫学与寄生虫病杂志

ABSTRACT

Objective To obtain the recombinant protein of an antigen gene Ts88 of Trichinella spiralis and identify the characteristics of the recombinant protein. Methods Ts88 cDNA obtained by immunoscreening the cDNA library of adult T. spiralis was subcloned into the pET 28c(+) expression vector and expressed in E.coli . Mice were immunized with the fusion protein incorporated into Freund’s adjuvant and the immune sera were collected. The titers of the Ts88 immune sera and the antigenicity of the recombinant protein were detected by ELISA and Western blotting. Immuno fluorescence test was performed in order to confirm the distribution of Ts88 protein in the worm. Results The fragment of Ts88 gene was expressed successfully in E.coli and a highly purified fusion protein was obtained. Immunization with the recombinant protein in mice produced high titers of antibodies, which recognized some components of native antigens of soluble proteins from adult worm of T. spiralis. Western blotting analysis showed that Ts88 recombinant antigen was recognized by all the positive sera, such as the sera from infected or immunized rabbits, from infected swine and from patients of trichinosis. Immuno fluorescence test confirmed that Ts88 protein mainly distributed in the cuticle surface of the worm. Conclusion The Ts88 antigen gene from T. spiralis was successfully expressed. The recombinant protein presented antigenicity.