牛抑制素pET32a-matpeptide重组蛋白质的诱导表达及鉴定  

Induced Expression and Identification of Recombinant Protein of Bovine Inhibin pET32a-matpeptide

作　　者：吴迪[1] 穆秋玲 齐晓龙[1] 常迪[1] 李贺娟[1] 郭勇[1] 倪和民[1] 

机构地区：[1]北京农学院动物科学技术学院兽医学(中医药)北京市重点实验室,北京102206

出　　处：《中国农学通报》2017年第14期121-124,共4页Chinese Agricultural Science Bulletin

基　　金：2013年度北京市教委北京市属高等学校创新团队建设与教师职业发展计划项目"体细胞转基因克隆肉牛新品系培育与利用"(IDHT20130515);奶牛产业体系北京市创新团队

摘　　要：为表达纯化出免疫原性良好的牛Inhibin pET32a-matpeptide重组蛋白质,利用IPTG对重组蛋白进行诱导表达,并利用重组蛋白大量表达纯化出目的蛋白,通过Western blot鉴定目的蛋白的免疫原性。结果表明:使用1 moL/L IPTG在18℃诱导表达18 h可得到重组抑制素蛋白,经重组蛋白小量表达确定目的蛋白以可溶性形式存在,Western blog试验表明重组蛋白有较高的免疫原性。本试验诱导表达出可溶性、免疫原性较高的抑制素pET32a-matpeptide重组蛋白质,为后续制备抗抑制素α亚基抗体奠定了基础。To express and purify recombinant protein of bovine inhibin pET32a-matpeptide with good immunogenicity, we used IPTG for inducible expression of recombinant protein. Then large amount of target protein were expressed and purified using the recombinant protein and Western blot was conducted to identify the immunogenicity of target protein. The results showed that recombinant inhibin protein could be induced and expressed using 1 moL/L IPTG at 18℃ for 18 h. Expression of a small amount of recombinant proteins proved the soluble form of the target protein. Western blot test showed that recombinant protein had higher immunogenicity. The soluble recombinant protein of pET32a-matpeptide with high immunogenicity induced in this experiment laid a foundation for the subsequent preparation of inhibin pET32a-matpeptide antibody.

关 键 词：牛抑制素 重组蛋白 蛋白免疫印迹 免疫原性 

分 类 号：Q812[生物学—生物工程]