高原鼠兔HIF-1α基因真核表达载体的构建及细胞表达和定位  

Construction of Eukaryotic Expression Vector of HIF-1α Gene in Plateau Pika and Its Expression and Localization in Cell

作　　者：李红阁[1,2] 赵新全[1] 常智杰[3] 任永明[3] 王德鹏[1,2] 

机构地区：[1]中国科学院西北高原生物研究所高原生物适应与进货重点实验室,青海西宁810001 [2]中国科学院研究生院,北京100049 [3]清华大学医学院生物科学与技术系生物膜与膜生物工程国家重点实验室,北京100084

出　　处：《安徽农业科学》2009年第10期4386-4388,4393,共4页Journal of Anhui Agricultural Sciences

基　　金：国家基础研究规划项目(2005CB422005)

摘　　要：利用PCR从克隆质粒PMD18-T-pHIF-1α中扩增鼠兔HIF-1α;构建重组质粒pcDNA3.0/6Xmyc-p HIF-1α,用重组质粒转染HEK-293T,HeLa和COS-7细胞,通过免疫印迹分析,检测HIF-1α表达蛋白在细胞中的分布。EcoRI和XhoⅠ酶切鉴定和序列分析表明,构建的pcDNA3.0/6Xmyc-pHIF-1α真核表达载体序列正确;HIF-1α在真核细胞中能够正确表达,且其表达产物主要存在于细胞核中。HIF-1α in cloning plasmid PMD18-T-pHIF-1α was amplified by PCR,and the recombinant plasmid pcDNA3.0/6Xmyc-pHIF-1α was constructed and was transfected into HEK-293T,Hela and COS-7 cells.The distribution of the protein expressed by HIF-1α in cells was detected by western blot analysis.The results of enzyme digestion and sequence analysis showed that the sequence of the constructed eukaryotic expression vector was correct.The HIF-1α gene could correctly expressed in eukaryotic cells and its expression products mainly exist in cell nuclear.

关 键 词：高原鼠兔 HIF-1Α基因 真核表达载体 

分 类 号：S821.2[农业科学—畜牧学]