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Construction of Efficient Expression System of Clostridium perfringens Epsilon Toxin Protein and Analysis of its Immune Protection

LU Tian-fen1, CHAI Tong-jie1, SUN Ling-yu1, LI Xin-xian1, HAO Hai-yu1, LV Chang-hui1, YANG Yan-wei2   

  1. 1. College of Animal Science and Technology, Shandong Agricultural University, Tai’an 271000, China;
    2. Animal Husbandry Bureau of Zouping, Binzhou 256600, China
  • Received:2012-03-12 Online:2012-09-20 Published:2012-09-18

Abstract: The epsilon toxin produced by type B and D Clostridium perfringens, could lead to fatal illness in a variety of livestock animals, causing financial loss. In this study, pET-28a(+) plasmid harboring epsilon toxin gene was transferred into E.coli strain BL21(DE3)to efficiently produce epsilon toxin.Results of SDS-PAGE and Western blotting showed that recombinant E.coli BL21(DE3) could highly expressed ε toxin (about 32.83% of total cellular protein). On this basis, rabbits were immunized with 0.2 mg of the insoluble recombinant protein to evaluate immune protection of the expressed epsilon protein. Results of indirect ELISA demonstrated that anti-ε antibody titer reached the maximum value, 8.7log2 in the fourth week. In conclusion, the recombinant strain BL21(DE3) could highly express ε toxin, and the toxin had excellent immune protection.

Key words: Clostridium perfringens; epsilon toxin; expression; immune protection

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