尼罗罗非鱼TGF-β1蛋白原核表达及多克隆抗体的制备  

Prokaryotic expression and polyclonal antibodies preparation of TGF-β1 protein in Nile tilapia

作　　者：余艳玲 冯鹏霏 潘传燕 陈晓汉 林勇 张永德 罗洪林 YU Yan-ling;FENG Peng-fei;PAN Chuan-yan;CHEN Xiao-han;LIN Yong;ZHANG Yong-de;LUO Hong-lin(Guangxi Key Laboratory for Aquatic Genetic Breeding and Healthy Aquaculture,Guangxi Academy of Fishery Sciences,Nanning 530021,China)

机构地区：[1]广西水产科学研究院广西水产遗传育种与健康养殖重点实验室

出　　处：《江苏农业学报》2019年第6期1407-1412,共6页Jiangsu Journal of Agricultural Sciences

基　　金：国家自然科学基金面上项目(31760765、31372553);广西自然科学基金项目(2015GXNSFAA139068)

摘　　要：为制备尼罗罗非鱼TGF-β1多克隆抗体,采用同源重组技术构建尼罗罗非鱼pET-B2m-TGF-β1原核表达载体,转入大肠杆菌B21中诱导表达,将重组表达蛋白质纯化后免疫大耳兔制备多克隆抗体,采用Western Blot和ELISA检测抗体的特异性和效价。结果表明,构建的pET-B2m-TGF-β1原核表达载体经诱导表达获得了分子量为5.2×104的重组蛋白质,免疫大耳兔获得效价为1∶2048000的抗尼罗罗非鱼TGF-β1多克隆抗血清,该抗体能够特异性地识别原核表达的TGF-β1蛋白。To prepare the polyclonal antibody against TGF-β1 of Nile tilapia,the TGF-β1 gene of Nile tilapia was transferred into the prokaryotic expression vector pET-B2m using homologous recombination technology,and transferred into Escherichia coli B21 for expression.The expressed TGF-β1 protein was purified and used to immunize rabbits for the preparation of polyclonal antibodies.The specificity and potency of the antibody were detected by Western blot and ELISA.The results showed that the pET-B2m-TGF-β1 prokaryotic expression vector was successfully constructed,and the recombinant protein with the molecular weight of 5.2×10^4 was obtained by inducing expression.The anti-Nile tilapia TGF-β1 polyclonal antiserum with a potency of 1∶2048000 was obtained from the immunized rabbit.The antibody could specifically recognize the prokaryotically expressed TGF-β1 protein.

关 键 词：罗非鱼 TGF-Β1蛋白 原核表达 多克隆抗体 

分 类 号：S965.125[农业科学—水产养殖]