Immunogenicity and Immunoprotection of Translocation Protein B(TolB)in Vibrio alginolyticus

doi:10.13560/j.cnki.biotech.bull.1985.2015.07.021

Abstract

Abstract:

To investigate the possibility of translocation protein B(TolB)from Vibrio alginolyticus HY9901 as a candidate antigen for vaccine production, the full length tolB gene was amplified by PCR and designed specific primers according to the whole genome sequence of V. alginolyticus published in GenBank. Sequence analysis revealed that tolB gene was 1 353 bp and encoded a putative protein of 450 amino acids(GenBank accession number：JQ846501). BLAST analysis showed that the tolB of V. alginolyticus had high genetic relationship with other known Vibrio, its sequence was conservative, and might be a common antigen candidate protein. The TolB protein of prokaryotic expression was injected into SPF mice to prepare polyclonal antibody, and its titer of ELISA reached 1∶40 000. Western blot indicated that TolB serum reacted specifically with the induced recombinant protein. Further the TolB protein was used as an antigen to immunize grouper(Epinephelus awoara)twice, and ELISA detection found that the antibody titer of the immunized grouper increased to the highest(up to 1∶2 048)in the fourth week. Challenge assay test showed that the immunoprotection rate of TolB to E. awoara was 76%. Therefore the TolB of V. alginolyticus has solid immunogenicity and immunoprotection, and it can be used as a candidate antigen for vaccine of Vibrio subunits.

Key words: Vibrio alginolyticus, tolB gene, immunogenicity, immunoprotection

Pang Huanying, Zhou Zejun, Zhang Yanfei, Li Jing, Qiu Mingsheng, Ding Yu, Jian Jichang, Wu Zaohe. Immunogenicity and Immunoprotection of Translocation Protein B(TolB)in Vibrio alginolyticus[J]. Biotechnology Bulletin, 2015, 31(7): 143-149.

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