文献类型：期刊文献

英文题名：Secretory expression of recombinant proteins in an attenuated Vibrio anguillarum strain for potential use in vaccines

作者：Shao, M; Ma, Y; Liu, Q; Zhang, Y

机构：[1]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2005

卷号：28

期号：12

起止页码：723

外文期刊名：JOURNAL OF FISH DISEASES

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000234345800005)】；

语种：英文

外文关键词：AngE-HlyAs; Escherichia coli alpha-haemolysin; GFP-HlyAs; protein secretion; vaccine; Vibrio anguillarum

摘要：Vibrio anguillarum is an important bacterial fish pathogen responsible for epizootics in both marine and freshwater fish worldwide. Studies on pathogenic V anguillarum has shown that its virulence is mediated by a 65 kb endogenous pJM1-like plasmid, which encodes an efficient iron uptake system. The plasmid-free derivative of wild V. anguillarum was found to be greatly attenuated and elicited a good protection against wild V anguillarum in fish. In this study, a plasmid-free derivative MVAV6201, an effective live vaccine candidate, was used as a carrier strain to achieve the secretory delivery of recombinant proteins in V anguillarum. The secretion mechanism was based on the Escherichia coli alpha-hacinolysin (HlyA) transport system. The recombinant proteins were fused with the alpha-haemolysin secretion signal (HlyAs) and expressed from the commonly used HlyA secretion vector pMOh-ly1. Two HlyAs-tagged recombinant proteins, GFP-HlyAs and AngE-HlyAs, were constructed and their secretion characters in V anguillarum investigated. In the case of GFP-HlyAs, nearly 70% of the total fusion protein was efficiently secreted into culture supernatant, and in the case of AngE-HlyAs, the secretion efficiency was determined to be about 300 mu g L-1 by Western blotting.