大口黑鲈蛙虹彩病毒MCP蛋白多克隆抗体的制备与应用  

Preparation and application of polyclonal antibody against MCP protein of largemouth bass ranavirus

作　　者：王若瑄 罗霞[2] 李宁求[2] 林强[2] 牛银杰 梁红茹[2] 付小哲[2] 吕爱军 WANG Ruoxuan;LUO Xia;LI Ningqiu;LIN Qiang;NIU Yinjie;LIANG Hongru;FU Xiaozhe;LÜAijun(College of Fisheries,Tianjin Agricultural University,Tianjin 300384,China;Pearl River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Key Laboratory of Fishery Drug Development,Ministry of Agriculture and Rural Affairs,Key Laboratory of Aquatic Animal Immune Technology,Guangdong Provinces,Guangzhou,Guangdong 510380,China)

机构地区：[1]天津农学院水产学院,天津300384 [2]中国水产科学研究院珠江水产研究所,农业农村部渔用药物创制重点实验室,广东省水产动物免疫与绿色养殖重点实验室,广东广州510380

出　　处：《西北农林科技大学学报（自然科学版）》2024年第4期21-27,共7页Journal of Northwest A&F University(Natural Science Edition)

基　　金：广东省海洋经济发展(海洋六大产业)专项资金项目(GDOE[2022]A13);中国水产科学研究院中央级公益性科研院所基本科研业务费专项(2020TD45);广东省现代农业产业技术创新团队专项基金项目(2019KJ140,2019KJ141)。

摘　　要：【目的】制备大口黑鲈蛙虹彩病毒主衣壳(MCP)蛋白兔多克隆抗体,以期为该病毒蛋白功能研究奠定基础。【方法】对pET32a(+)MCP/BL21重组大肠杆菌进行诱导表达,将重组蛋白进行纯化、复性,以此为抗原免疫大耳兔制备多克隆抗体,ELISA检测抗体效价,间接免疫荧光试验(IFA)和Western Blot法分析MCP多克隆抗体的特异性。【结果】纯化的MCP重组蛋白条带特异;间接ELISA结果显示,制备的兔多克隆抗体血清效价为1∶1024000;IFA和Western Blot检测结果表明该多抗特异性良好,能够与大口黑鲈蛙虹彩病毒MCP蛋白发生特异性反应,IFA试验表明血清最适稀释度为1∶500。【结论】成功制备了大口黑鲈蛙虹彩病毒MCP兔多克隆抗体,该抗体可特异性识别大口黑鲈蛙虹彩病毒MCP蛋白。【Objective】The rabbit polyclonal antibody against recombinant MCP protein of largemouth bass ranavirus(LMBV)were prepared to provide basis for investigating LMBV protein functions.【Method】The pET32a(+)MCP/BL21 recombinant E.coli was induced to express,and the recombinant protein was purified and renatured.The polyclonal antibody was prepared by immunizing rabbits with the recombinant protein.The antibody titer was detected by ELISA and the specificity of MCP polyclonal antibody was analyzed by indirect immunofluorescence assay(IFA)and Western Blot.【Result】The purified MCP recombinant protein was specific.Indirect ELISA results showed that the serum titer of rabbit polyclonal antibody was 1∶1024000.The results of IFA and Western Blot illustrated polyclonal antibody had good specificity to LMBV.The optimum dilution of serum in IFA test was 1∶500.【Conclusion】The poly-clonal antibody against MCP of LMBV was successfully prepared with specific recognition of major capsid proteins of largemouth bass ranavirus.

关 键 词：大口黑鲈 蛙虹彩病毒 主衣壳蛋白 多克隆抗体 

分 类 号：S943.211.417[农业科学—水产养殖]