重组人干细胞因子在大肠杆菌中的表达及活性鉴定  

Expression of Recombinant Human Stem Cell Factor in E.coli and its Biological Activity

作　　者：谭文斌[1,2] 成光杰[1,2] 李卫民 谭运年[1,2] 朱定尔 

机构地区：[1]湖南医科大学分子生物学研究中心 [2]湖南医科大学血液生理研究室

出　　处：《生命科学研究》1999年第1期48-52,共5页Life Science Research

基　　金：国家自然科学基金

摘　　要：采用ＰＣＲ方法从重组载体ｐＫＫＳＣＦ扩增出人干细胞因子（ＳＣＦ）基因ｃＤＮＡ片段，经亚克隆构建成表达载体ｐＥＴ１１ｄＳＣＦ，经限制性酶谱分析鉴定证实．继用ＩＰＴＧ诱导重组ｐＥＴ１１ｄＳＣＦ在大肠杆菌ＨＭＳ１７４（ＤＥ３）中表达，经初步纯化获得重组人ＳＣＦ蛋白质，经检测具有生理活性，对脐带血ＣＦＵ－ＣＭ和ＨＰＰ－ＣＦＣ增殖有明显刺激作用，为进一步的生物工程研究奠定了基础．Abstract A cDNA fragment of human stem cell factor (SCF) was amplified from the recombinant plasmid of pKKSCF by PCR and subcloned into an expression vector of pET11dSCF which was examined by restriction enzyme mapping. And then it was transfected into E. coli strain HMS174(DE 3). The bacteria containing the recombinant plasmid of pET 11 dSCF, could express the recombinant SCF (rSCF) protein by induction with IPTG in culture. A larger yield of rSCF in HMS174 (DE 3), particularly in the bacteria inclusion bodies was observed. The rSCF isolated from cultured E.coli waspartlypurified

关 键 词：干细胞因子 基因表达 生物工程 重组 

分 类 号：Q78[生物学—分子生物学] Q516