Pearl River Fisheries Research Institute, CAFS, Guangdong Key Laboratory of Aquatic Animal ImmunoTechnology, Guangzhou 510380, PR China.
Fish Shellfish Immunol. 2010 May-Jun;28(5-6):952-6. doi: 10.1016/j.fsi.2010.02.010. Epub 2010 Feb 17.
The outer membrane protein-OmpK has been considered as a vaccine candidate for the prevention of infections due to Vibrio harveyi, Vibrio alginolyticus and Vibrio parahaemolyticus in fish. Interestingly, the polyclonal antibody raised against the recombinant OmpK from V. harveyi strain EcGs020802 recognized the OmpK homologues from other strains of Vibrio species by immunoblotting. The ompK genes from 19 Vibrio strains including V. harveyi (11), V. alginolyticus (6) and V. parahaemolyticus (2) were then cloned and sequenced. Alignment analysis based on the amino acid sequences indicated that the OmpK from V. harveyi strain EcGs020802 had 71.7-99.2% of identities with those from V. harveyi, V. alginolyticus and V. parahaemolyticus. Western blot analysis revealed that the corresponding native proteins ranged between 28 and 31 kDa, consistent with predicated molecular weight of OmpK in Vibrio strains. Furthermore, the cross-protective property of recombinant OmpK was evaluated through challenge with heterogeneous virulent Vibrio strains in Orange-spotted groupers (Epinephelus coioides). Orange-spotted groupers vaccinated with recombinant OmpK were more tolerant of the infection by virulent Vibrio strains and their relative percentage survival (RPS) was correlative with the degree of the identity of deduced amino acid sequences of their OmpK. Taken together, the OmpK is a conserved protective antigen among tested Vibrio species and might be a potentially versatile vaccine candidate for the prevention of infections due to V. harveyi, V. alginolyticus and V. parahaemolyticus.
外膜蛋白-OmpK 已被认为是预防鱼类感染哈维弧菌、溶藻弧菌和副溶血弧菌的疫苗候选物。有趣的是,针对哈维弧菌 EcGs020802 株重组 OmpK 产生的多克隆抗体通过免疫印迹识别了其他弧菌属菌株的 OmpK 同源物。然后克隆和测序了来自 19 株弧菌的 ompK 基因,包括哈维弧菌(11 株)、溶藻弧菌(6 株)和副溶血弧菌(2 株)。基于氨基酸序列的比对分析表明,哈维弧菌 EcGs020802 株的 OmpK 与哈维弧菌、溶藻弧菌和副溶血弧菌的同源物具有 71.7-99.2%的同一性。Western blot 分析显示,相应的天然蛋白大小在 28 至 31 kDa 之间,与预测的弧菌菌株 OmpK 的分子量一致。此外,通过用异源毒性弧菌菌株挑战橙斑石斑鱼(Epinephelus coioides)评估了重组 OmpK 的交叉保护特性。用重组 OmpK 接种的橙斑石斑鱼对毒性弧菌菌株的感染更耐受,其相对存活率(RPS)与推导的 OmpK 氨基酸序列的同一性程度相关。总之,OmpK 是测试的弧菌属中保守的保护性抗原,可能是预防哈维弧菌、溶藻弧菌和副溶血弧菌感染的潜在多功能疫苗候选物。