摘要
采用大肠杆菌表达体系来获得陆地棉锌指蛋白(GZFP)的融合蛋白.以pMD18-GZFP质粒为模板,用PCR方法扩增获得GZFP基因的编码区序列,将其克隆到表达载体pET28b(+)中,转化宿主菌BL21(DE3),成功地构建了陆地棉GZFP基因原核表达载体pET-GZFP、经IPTG诱导、SDS-PAGE电泳分析和蛋白质杂交检测表明,陆地棉GZFP以融合蛋白的形式在大肠杆菌中得到大量表达.
In this study, a novel zinc finger protein （GZFP） from upland cotton （Gossypiurn hirsutum L. ） was expressed by the E. coli expression system. The GZFP gene was amplified by PCR with pMD18-GZFP plasmid as the template, and was then cloned into the pET-28b（＋） to construct pET-GZFP recombinant plasmid, which was transformed into E. coli BL21（DE3）. The recombinant GZFP was induced and expressed by IPTG. SDS-PAGE analysis and Western blotting showed that the target protein was expressed at a high level in E. coli BL21 （DE3）.
出处
《西南大学学报（自然科学版）》
CAS
CSCD
北大核心
2007年第3期130-133,共4页
Journal of Southwest University(Natural Science Edition)
基金
国家自然科学基金资助项目(30440032)
重庆邮电大学博士启动基金资助项目(A2005-13)
关键词
锌指蛋白
原核表达
陆地棉
zinc finger protein
prokaryotic expression
Gossypiurn hirsutum L.
