期刊
JOURNAL OF FOOD SCIENCE
卷 75, 期 4, 页码 M198-M203出版社
WILEY
DOI: 10.1111/j.1750-3841.2010.01565.x
关键词
cloning; expression; genus-specificity; OmpK; polyclonal antiserum; Vibrio alginolyticus
资金
- National Natural Science Foundation of China [30371059]
- Xiamen Science and Technology Bureau, China [3502Z20055009]
Some Vibrio species are universal marine pathogens and Vibrio infections are often encountered due to consumption of raw or uncooked seafood. The outermembrane proteins, playing a key role in interaction between bacteria and hosts, are potential candidates for development of vaccine and markers of the genus Vibrio. In this study, the ompK (outer membrane protein K) genes of Vibrio alginolyticus, V. vulnificus, V. parahaemolyticus, V. fluvialis, and V. mimicu were cloned with 798 to 822 nucleotides encoding 266 to 274 amino acids. The ompK gene from V. alginolyticus was expressed in Escherichia coli using pET-22b expression vector. The recombinant fusion OmpK protein with 6xHis tag was purified with nickel chelate affinity chromatography. The polyclonal antibody (titer, 1:102400) against V. alginolyticus OmpK was developed in guinea pigs and it positively reacted with each of 5 Vibrio species but negative to other 18Gram-negative bacterial strains. The result suggests that Vibrio OmpK protein could be a genus-specific antigen, which can be used for developing vaccines and rapid detection of multiple Vibrio species.
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