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迟缓爱德华氏菌OppA蛋白的免疫原性及免疫保护分析
王妍妍1,2, 李贵阳1,2, 李 杰1,2, 肖 鹏1, 莫照兰1
1.中国科学院海洋研究所实验海洋生物学重点实验室;2.中国科学院研究生院
摘要:
迟缓爱德华氏菌(Edwardsiella tarda)是水产养殖动物重要的病原菌, 引起鱼类爱德华氏菌病, 免疫防治是针对该病的一个有效途径。OppA 是迟缓爱德华氏菌的一个寡肽透过酶组分, 作者开展OppA的免疫原性及免疫保护性的研究, 旨在评价其作为鱼类爱德华氏菌病疫苗候选抗原的可行性。将致病性的迟缓爱德华氏菌LSE40 的oppA克隆于表达载体pPROEXHTa, 转化入大肠杆菌BL21(DE3), 表达重组蛋白6His-OppA, 并利用Ni-NTA SefinoseTM kit 进行纯化。以纯化的6His-OppA 两次注射免疫大菱鲆(Scophthalmus maximus), 第34 天后检测血清抗体效价为1: 1024; 以LSE40 对免疫的大菱鲆进行人工感染, 测得免疫保护率是25.9%。结果表明, OppA 可刺激大菱鲆产生免疫应答, 并具有免疫保护效果。
关键词:  迟缓爱德华氏菌(Edwardsiella tarda)  OppA  原核表达  免疫应答  免疫保护
DOI:
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基金项目:国家自然科学基金项目(31072245); 国家鲆鲽类产业技术体系项目(NYCYTX-50); 青岛市科技发展计划项目(08-1-7-2-HY)
Immunogenicity and immunoprotection of oligopeptide permease A (OppA) of Edwardsiella tarda
Abstract:
Edwardsiella tarda is an important pathogen to aquaculture animals, causing fish edwardsiellosis diseases. Vaccination is an efficient protection against this disease in fish. OppA is a component of the oligopeptide permease system of E. tarda. We evaluated the potential of OppA as an antigenic candidate vaccine against edwardsiellosis. The oppA gene was cloned into expression vector pPROEXHTa and expressed in E. coli BL21 (DE3). The expressed 6His-OppA was purified with Ni-NTA SefinoseTM Kit and used as an antigen to immunize turbot (Scophthalmus maximus) twice. At day 34 post-vaccination, the serum antibody titer in the immunized fish was 1:1024, and the relative percentage survival (RPS) was 25.9% when the immunized turbot was challenged with pathogenic E. tardaa. The present studies indicate that OppA of E. tarda can induce immune response in fish and provide protection against edwardsiellosis.
Key words:  Edwardsiella tarda  OppA  Prokaryotic expression  Immune response  Immunoprotection
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