摘要
将扩增的鸡γ-干扰素基因(CHIFN-γ)克隆至原核表达载体pET30a上,构建了重组表达质粒pET-30a-CHIFN-γ。将其转入大肠杆菌BL21(DE3)中,于37℃不同时间诱导表达,经SDS-PAGE分析表明该蛋白在大肠杆菌中获得了高效表达,且以可溶蛋白的形式存在。蛋白通过镍离子亲和树脂进行纯化,并作为免疫原制备兔抗CHIFN-γ多抗血清。Western-blotting结果表明目的蛋白与多抗血清有免疫反应性。
The cDNA of chicken IFN-γ gene was cloned into pET30a vector to construct recombinant plasmid pET- 30a-CHIFN-γ. The recombinant plasmid was transformed into E.coli BL21 （DE3）and the bacterium was induced with IPTG. It was demonstrated by SDS-PAGE that the soluble protein was expressed in E.coli BL21 （DE3）. This protein was purified by Ni-NTA and used to prepare the antibody against chicken IFN-γ protein. The Western blotting result showed that the antibody could react with purified recombinant chicken IFN-γ protein.
出处
《中国家禽》
北大核心
2010年第1期20-22,26,共4页
China Poultry
基金
黑龙江省自然科学基金重点项目资助(ZNJ0702-01)
关键词
鸡γ-干扰素
表达
多抗血清
chicken interferon-gamma
expression
antiserum
