期刊
LETTERS IN APPLIED MICROBIOLOGY
卷 53, 期 1, 页码 22-29出版社
WILEY
DOI: 10.1111/j.1472-765X.2011.03062.x
关键词
DNA vaccine; immunity; outer membrane protein; turbot; Vibrio harveyi
资金
- National High-Tech. RD Programme [2007AA09Z416]
- Natural Science Foundation of China [30972275]
- Ministry of Agriculture of the People's Republic of China [nyhyzx 07-046]
Aims: The main aims of this study were to clone and express a new outer membrane protein U (OmpU) from a pathogenic Vibrio harveyi SF-1 and investigate its immune efficiency as a vaccine candidate against V. harveyi infection in turbot (Scophthalmus maximus). Methods and Results: In this study, a new gene, ompU was cloned from the genomic DNA of pathogenic V. harveyi SF-1. The ompU gene encoded a 35 kDa protein, which was purified by Ni-NTA His-Bind Resin column. A DNA vaccine was constructed by inserting ompU gene into pEGFP-N1 plasmid. Turbot were injected intramuscularly with the purified OmpU protein and the recombinant pEGFP-N1/ompU plasmid, respectively. The fish vaccinated with the purified OmpU protein were completely protected with a relative per cent of survival (RPS) of 100% against pathogenic V. harveyi infection. Efficient protection was also found in the pEGFP-N1/ompU vaccinated group, with a RPS of 51.4%. Significant specific antibody responses were detected in the vaccinated turbot by indirect enzyme-linked immunosorbent assay. Conclusions: A new OmpU was cloned and expressed. Both OmpU protein vaccine and DNA vaccine showed good immune protections in turbot. Significance and Impact of the Study: The OmpU was identified to be a new effective vaccine candidate and could be used as subunit vaccine and DNA vaccine for disease control caused by pathogenic V. harveyi.
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