绵羊BST-2B基因的原核表达及多克隆抗体的制备  

Prokaryotic expression and polyclonal antibody preparation of BST-2B gene in ovis aries

作　　者：刘腾[1] 张莉 谭永贵 郭慧敏[1] 朱杰[1] 缪秋红[1] 李传峰[1] 陈宗艳[1] 刘光清[1] 

机构地区：[1]中国农业科学院,上海兽医研究所,上海200241

出　　处：《中国兽医科学》2017年第11期1406-1410,共5页Chinese Veterinary Science

基　　金：国家重点研发计划项目(2016YFD0500108);国家自然科学基金项目(31502068);上海市科技兴农重点攻关项目(2016043);公益性农业科研专项(201303046);中央级公益性科研院所基本科研业务费专项(2016JB01)

摘　　要：为了表达含有绵羊BST-2B(o BST-2B)基因的重组蛋白,用RT-PCR方法扩增绵羊肺细胞的o BST-2B基因,并将其克隆至原核表达载体p GEX-4T-1中,构建重组表达质粒p GEX-4T-o BST-2B。将重组表达质粒转化至大肠杆菌BL21(DE3),用IPTG诱导表达重组o BST-2B蛋白。将表达产物纯化后免疫BALB/c小鼠,制备抗o BST-2B的多克隆抗体。蛋白免疫印迹分析和间接免疫荧光试验检测结果显示,该抗体具有较好的反应原性。本试验成功制备的o BST-2B蛋白及多克隆抗体,为研究o BST-2B蛋白的生物学功能及羊传染病的致病机制提供了良好的基础材料。In order to express recombinant protein of BST-2 B gene from Ovis aries,o BST-2 B gene was amplified from sheep lung cells by RT-PCR and cloned the fragment into p GEX-4 T-1 vector to construct p GEX-4 T-o BST/2 B recombinant expression plasmid.The recombinant plasmid was transformed into E.coli BL21（DE3）and induced with IPTG.The recombinant protein was purified and used to immunize BALB/c mice to obtain a polyclonal antibodies against o BST-2 B.Western-blot analysis and indirect immunofluorescence assay showed that the polyclonal anti-bodies had a specifical immunogenicity.The o BST-2 B protein and polyclonal antibodies may provide a basal bio-material for researches of sheep diseases and the biofunctions of o BST-2 B protein.

关 键 词：BST-2B基因 原核表达 多克隆抗体 

分 类 号：S852.4[农业科学—基础兽医学]