【摘 要】
：
创伤弧菌(Vibrio vulnificus)是欧洲鳗鲡(Anguilla anguilla)出血性败血症的重要致病菌,严重危害了欧鳗的养殖.本实验室于2003年7月从福建省长乐市发病欧洲鳗鲡上分离到毒力较强的创伤弧菌菌株FJ03-X2.为减小创伤弧菌对欧鳗养殖业的危害,研究开发具有较好免疫保护力的核酸疫苗,本实验以此菌株为病原菌,选出毒力因子ompk为目的基因,构建DNA疫苗.首先,构建pET3
【机 构】
：
福建省农业科学院生物技术研究所 福州 350003;福建农林大学动物科学学院 福州 350002
【出 处】
：
2013海水养殖鱼类病原与病毒控制工程论坛
论文部分内容阅读
创伤弧菌(Vibrio vulnificus)是欧洲鳗鲡(Anguilla anguilla)出血性败血症的重要致病菌,严重危害了欧鳗的养殖.本实验室于2003年7月从福建省长乐市发病欧洲鳗鲡上分离到毒力较强的创伤弧菌菌株FJ03-X2.为减小创伤弧菌对欧鳗养殖业的危害,研究开发具有较好免疫保护力的核酸疫苗,本实验以此菌株为病原菌,选出毒力因子ompk为目的基因,构建DNA疫苗.首先,构建pET32a-OMPK原核表达质粒,在大肠杆菌BL21中诱导表达,纯化重组蛋白,免疫SD大鼠获得血清抗体,经ELISA检测血清效价达到1：100000.同时,构建真核表达载体pcDNA3-OMPK,转染293细胞,经RT-PCR检测,得到大小相符的条带,表明重组质粒在293细胞中有表达.将pcDNA3-OMPK质粒肌肉注射免疫模式生物斑马鱼,7天后,通过RT-PCR成功检测到质粒在斑马鱼肌肉组织中的表达,为后续检测DNA疫苗的免疫保护力等实验打下基础.
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