核心链霉亲和素基因的克隆及表达  

Cloning and Expression of the Core-Streptavidin Gene in E.coli

作　　者：李彪[1] 杨卫东[1] 朱承谟[1] 吴祥甫[2] 

机构地区：[1]上海第二医科大学附属瑞金医院核医学科,上海200025 [2]中国科学院上海生化研究所,上海200031

出　　处：《上海免疫学杂志》2003年第3期194-196,200,共4页Shanghai Journal of Immunology

基　　金：国家自然科学基金资助项目 (No 3 9770 2 3 1);上海市启明星计划资助项目 (编号 :98QB14 0 11)

摘　　要：文章采用PCR方法 ,以链霉亲和素菌 (Streptmycesavidinii)基因组DNA为模板 ,克隆出 384bp的DNA片段并以BamHI及XhoI位点装入PSK载体 ,经测序确证为核心链霉亲和素基因 (core strepavidin )。将该基因重组到质粒pET 2 4a ( + ) ,构建重组表达质粒pET 2 4a CS。转化大肠杆菌BL2 1,经IPTG诱导后得到高效表达。SDS PAGE图谱显示表达产物相对分子质量为15 0 0 0 ,与其基因编码蛋白的理论值相符。HRP标记的生物素作为抗体进行Westernblot,结果在 15 0 0 0处可见特异性条带 。Core streptavidin gene has been cloned from Streptmyces avidinii using PCR amplication The amplified 384 bp DNA fragment was confirmed by DNA sequencing to be known corestreptavidin gene The fragment was insert into the vector pET 24a(+) to construct an expression plasmid pET 24a(+) CS The core streptavidin was highly expressed in E coli as inclusion bodies It was evidenced by SDS PAGE and Western blot Our result showed that the protein of core streptavidin can bind biotin specifically

关 键 词：克隆 表达 核心链霉亲和素基因 

分 类 号：R730.3[医药卫生—肿瘤]