猪生长激素基因的原核表达及其抗体制备  

Prokaryotic expression of porcine GH gene and its polyclonal antibody preparation

作　　者：李凤[1] 孟亚鹏[1] 刘燕飞[1] 谭俊杰[1] 付若彬[1] 刘世贵[1] 龙章富[1] 

机构地区：[1]四川大学生命科学学院

出　　处：《中国兽医科技》2005年第1期52-55,共4页Chinese Journal of Veterinary Science and Technology

基　　金：四川省应用基础研究项目(02NY029 079)

摘　　要：通过构建pGH基因的原核表达质粒,转化大肠埃希氏菌TOP10,经IPTG诱导,成功表达了重组猪生长激素的融合蛋白。经SDS PAGE分析,在分子质量50.4ku处有1条新的特异蛋白质条带。对以包涵体形式表达的融合蛋白进行SDS PAGE分离,并经透析得到了重组融合蛋白,以此融合蛋白免疫家兔,制备并纯化了抗pGH多克隆抗体,经酶联免疫吸附测定、蛋白质印迹分析和免疫组织化学方法检测,此抗体具有较高效价和特异性。成功地表达了pGH基因并获得了多克隆抗体。Expression vector of pGH gene was constructed by restrictive enzyme digestion and gene recombination techniques and was transformed into E.coli TOP10 for IPTG induced expression .The result showed that pGH fusion protein was expressed in the form of inclusion body. A (protein) band with Mw (50.4) ku appeared on SDS-PAGE. Fusion protein was isolated by SDS-PAGE and dialysis.New Zealand (rabbits) were immunized with pGH fusion protein to (prepare) polyclonal antibodies against pGH. The pGH (antiserum) was obtained and purified. It was characterized by ELISA,Western blotting, immnunohistochemistry and immuncyttochemistry. The (results) showed that the antibodies had high titer,affinity and specificity.

关 键 词：猪 生长激素 原核表达 多克隆抗体 

分 类 号：S859.793[农业科学—临床兽医学] Q786[农业科学—兽医学]