大鼠βB2-晶体蛋白的表达纯化及多克隆抗体的制备  

Expression and purification of rat βB2-crystallin and preparation of its polyclonal antibody

作　　者：胡书群[1] 裴冬生[1] 陆梁[1] 赵惠仁[1] 

机构地区：[1]徐州医学院生物化学与分子生物学研究中心,221002

出　　处：《免疫学杂志》2011年第5期433-436,共4页Immunological Journal

基　　金：江苏省自然科学基金面上项目(BK2010171);江苏省高校自然科学基金重点项目(10KJA310053);徐州医学院院长基金(2010KJZ12)

摘　　要：目的表达并纯化βB2-晶体蛋白,制备其多克隆抗体,用以研究βB2-晶体蛋白聚合的机制。方法用RT-PCR的方法扩增了βB2-晶体蛋白的cDNA并克隆到原核表达载体pMON5473中。用萘啶酮酸诱导recA启动子后,蛋白质在大肠杆菌中得到了高效表达。表达的蛋白质用DEAE纤维素层析和凝胶过滤纯化,SDS-PAGE显示一条带。与佐剂混合后免疫新西兰大白兔。结果纯化了大鼠βB2-晶体蛋白,成功制备了其多克隆抗体。结论制备的多克隆抗体为下阶段的研究提供了重要的实验材料。We aimed to express and purify the rat βB2-crystallin and to prepare the polyclonal antibody against the rabbit anti βB2-crystallin for studying the mechanism of their oligomerization.First,cDNA encoding rat βB2-crystallin was amplified using RT-PCR,and then cloned into the procaryotic expression vector pMON5473.After the induction of recA promoter with nalidixic acid,chromatography on DEAE cellulose,and gel filtration on Sephadex,the protein was obtained,which was confirmed by SDS-PAGE.Then,the purified protein was used to immune the rabbit and successfully prepared the rabbit anti-βB2-crystallin polyclonal antibody which can be used in further studies.

关 键 词：βB2-晶体蛋白 表达 纯化 抗体 

分 类 号：R392.1[医药卫生—免疫学]