小鼠Oct4基因真核表达载体的构建及表达鉴定  

Construction and identification of eukaryotic expression vector of mouse Oct4 gene

作　　者：陈天姬[1] 杜娟[1] 卢光琇[1] 

机构地区：[1]中南大学生殖与干细胞工程研究所,湖南长沙410078

出　　处：《现代生物医学进展》2010年第9期1610-1612,共3页Progress in Modern Biomedicine

基　　金：国家自然科学基金资助项目(30570937);中南大学研究生创新基金资助项目(2340-74334000003)

摘　　要：目的:构建小鼠Oct4基因真核表达载体并对其表达进行鉴定。方法:以小鼠胚胎干细胞(mouse embryonic stem cells,mESCs)cDNA为模板克隆Oct4基因编码区序列,将其通过定向克隆插入pcDNA3.1(+)载体多克隆酶切位点(multiple cloning sites,MCS)中,形成pcDNA3.1(+)/Oct4重组载体。将pcDNA3.1(+)空载体和pcDNA3.1(+)/Oct4重组载体分别转染293T细胞,通过Western Blotting检测Oct4蛋白的表达。结果:成功检测到Oct4蛋白的表达。结论:成功构建了小鼠Oct4基因真核表达载体。Objective：To construct a eukaryotic expression vector of mouse Oct4 gene and identify its expression.Methods：Taking the mouse embryonic stem cells（mESCs） cDNA as template,the encoding sequence of mouse Oct4 gene was cloned and then inserted into the multiple cloning sites（MCS） of pcDNA3.1（＋） vector by directional cloning.The pcDNA3.1（＋） vector and pcDNA3.1（＋） /Oct4 recombinant vector were transfected into 293T cells separately and the expression of Oct4 protein was detected by Western Blotting.Result：The Oct4 protein was detected successfully.Conclusion：A eukaryotic expression vector of mouse Oct4 gene was constructed successfully.

关 键 词：小鼠胚胎干细胞 Oct4基因 pcDNA3.1(+)载体 

分 类 号：Q953[生物学—动物学] Q291