Identification and evaluation of an outer membrane protein OmpU from a pathogenic Vibrio harveyi isolate as vaccine candidate in turbot (Scophthalmus maximus)

多宝鱼 生物 哈维氏弧菌 微生物学 菱鲆属 质粒 dna疫苗 重组DNA 病毒学 弧菌科 基因 弧菌 遗传学 细菌 渔业
作者
Q. Wang,J. Chen,R. Liu,Juntao Jia
出处
期刊:Letters in Applied Microbiology [Oxford University Press]
卷期号:53 (1): 22-29 被引量:45
标识
DOI:10.1111/j.1472-765x.2011.03062.x
摘要

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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