Immunoscreening and identification of a novel full-length cDNA encoding membrane protein of Trichinella spiralis

doi:10.3969/j.issn.1006-7795.2015.02.013

Abstract

Abstract: Objective In order to obtain vaccine or diagnostic candidate antigens for trichinellosis, the adult cDNA library of T. spiralis was immunoscreened. Methods The adult cDNA library of T. spiralis was screened using the swine sera infected with T. spiralis. The cDNA sequence of the positive clone was analyzed and the recombinant protein was expressed and purified. The antigenicity and immunogenicity of the recombinant protein was analyzed with ELISA and Western blotting. Results Forty-two positive clones were identified by immunscreened. A novel gene Tmp10 of T. spiralis was sequenced and recombinant plasmid pET-28a(+)/Tmp10 was successfully constructed. The recombinant protein(rTmp10) was expressed and the molecular weight was about 40 000. The results of Western blotting showed that the rTmp10 could be recognized by sera from the mice and swine infected with T. spiralis respectively, which showed that the rTmp10 had the specific antigenicity. The rTmp10 could induce a high level of specific anti-Tmp10 IgG antibodies, which showed that the rTmp10 had the immunogenicity. Conclusion The novel gene Tmp10 of T. spiralis was obtained by immunoscreening and predicted to be a membrane protein. The recombinant protein Tmp10 had the high specific antigenicity and immunogenicity.

Key words: Trichinella spiralis, cDNA library, membrane protein, expression

CLC Number:

R383.1

Bi Kuo, Qin Jiajia, Yang Jing, Zhu Xinping. Immunoscreening and identification of a novel full-length cDNA encoding membrane protein of Trichinella spiralis[J]. Journal of Capital Medical University, 2015, 36(2): 232-238.

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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2015.02.013

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2015/V36/I2/232

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