人颗粒裂解肽突变子在大肠埃希菌中的构建与表达研究  

Construction and expression of a fusion gene expression vector containing human NKG5-Ser

作　　者：赵铭 刘冰 王臻[1,2] 李邦印 张灵霞 

机构地区：[1]解放军第三〇九医院,北京100091 [2]江西农业大学,江西南昌330045

出　　处：《中国医药导报》2013年第14期26-28,35,共4页China Medical Herald

基　　金：首都医学发展科研基金(编号2002-3030)

摘　　要：目的构建NKG5突变基因的原核表达载体并进行表达研究。方法采用寡核苷酸合成的方法,拼接成为NKG5-Ser的编码序列,克隆到pGEMT载体上,测序正确后,再切下编码序列连接到质粒pGEX-4T-1中,重组表达载体转化大肠埃希菌BL21(DE3)pLysS,用IPTG诱导使融合蛋白高效表达。结果成功获得重组表达载体pGEX-4T-1-NKG5-Ser,转染大肠埃希菌BL21(DE3)pLysS,诱导表达后,SDS-PAGE和免疫印迹反应均显示显示出相对分子量(Mr)为35 kD的蛋白条带,证明融合蛋白表达成功。结论获得了突变NKG5与GST的融合蛋白,为进一步的工作打下了基础。Objective To construct prokaryotic expression vector carrying NKG5-Ser eDNA and to express it in E.coli. Methods The mutated NKGS, NKGS-Ser gene was obtained and cloned into clone vector pGEMT, it was sequenced and then cloned into expression vector pGEX-4T-1. Thus the prokaryotie expression vector pGEX-4T-1-NKG5-Ser was constructed, and E.coli BL21（DE3） pLysS was transformed, induced by IPTG to obtain a fusion protein. Results NKG5- Ser recombinant expression vector, pGEX-4T-1-NKG5-Ser was obtained. While E. coli BL21（DE3） pLysS carrying the expression vector was induced by IPTG, a fusion protein with relative molecular mass （Mr） 35 kD was detected by Western blot analysis. Conclusion A GST-NKG5-Ser fusion protein is obtained, so as to provides experiment basis for further research.

关 键 词：颗粒裂解肽 突变基因 融合蛋白 原核表达 

分 类 号：R978.3[医药卫生—药品]