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弧菌是海水养殖环境中常见的条件性致病菌,弧菌病的暴发给水产养殖业造成了严重损失。鉴于水生动物尤其是鱼类弧菌病的发生常常是多种(血清型或亚种)弧菌的混合感染,筛选具有潜在的交叉保护性蛋白抗原,作为制备多价疫苗或联合疫苗的侯选成分具有重要意义。文中从患病大黄鱼中分离到8株弧菌,经生理生化和分子生物学鉴定分别为6株哈维氏弧菌Vibrio harveyi,1株溶藻弧菌Vibrio alginolyticus和1株副溶血弧菌Vibrio parahaemolyticus。选择典型的不同种的弧菌为代表,提取其外膜蛋白,经SDS-PAGE和Westernblotting分析,确定它们大约在45 kDa、35 kDa、22 kDa处出现了3条共同的免疫印迹条带,表明它们很有可能含有共同的能够彼此交叉保护的抗原。利用双向电泳和免疫印迹相结合的方法,借助于MALDI-TOF-MS质谱分析技术,发现溶藻弧菌V.alginolyticus的一种功能未知的孔蛋白(Porin,GenBank Accession No.ZP_01260407)和副溶血弧菌V.parahaemolyticus的一种麦芽糖孔蛋白的前体蛋白(Maltoporin precursor,GenBank AccessionNo.NP_801154)能够和哈维氏弧菌V.harveyi全菌多抗产生免疫反应,表明这两种蛋白可以作为3种弧菌的交叉保护性抗原,以此制备的疫苗可望对3种弧菌的感染产生交叉保护作用。
Vibrio is a common opportunistic pathogen in the mariculture environment, and the outbreak of vibriosis caused serious losses to the aquaculture industry. As aquatic animals, especially fish vibriosis, are often mixed infections of multiple (serotypes or subspecies) vibrios, screening for potential cross-protective protein antigens as candidates for the preparation of multivalent or combination vaccines Composition is of great importance. Eight strains of Vibrio were isolated from the diseased large yellow croaker. Six strains of Vibrio harveyi, one strain of Vibrio alginolyticus and one strain of Vibrio parahaemolyticus were identified by physiological, biochemical and molecular biological methods. parahaemolyticus. By selecting typical Vibrio species as representative, the outer membrane proteins were extracted and analyzed by SDS-PAGE and Western blotting to determine that there were three common immunoblot bands at about 45 kDa, 35 kDa and 22 kDa They are likely to contain common antigens that cross-protect each other. Using a combination of two-dimensional electrophoresis and immunoblotting, MALDI-TOF-MS mass spectrometry was used to detect a porin (Porin, GenBank Accession No.ZP_01260407) and Vibrio parahaemolyticus A Maltoporin precursor (GenBank Accession No. NP_801154) from Vibrio V. parahaemolyticus is capable of producing an immunological response to whole virulence of V. harveyi, indicating that these two proteins are available as 3 Vibrio cross-protective antigen, the vaccine prepared by this is expected to cross-protect three strains of Vibrio infection.