| 人HCN2 真核表达载体的构建及在HEK293 细胞中的表达 |
| |
| 引用本文： | 左广锋 陈绍良 徐艳 肖杭. 人HCN2 真核表达载体的构建及在HEK293 细胞中的表达[J]. 现代生物医学进展, 2011, 11(6): 1068-1071 |
| |
| 作者姓名： | 左广锋 陈绍良 徐艳 肖杭 |
| |
| 作者单位： | 1. 南京医科大学附属南京市第一医院心内科,江苏,南京,210006
2. 南京医科大学江苏省医药动物实验基地,江苏,南京,210029 |
| |
| 摘 要： | 目的:构建含有人HCN2基因的真核表达载体,并观察在人胚胎肾细胞(HEK293)中的表达情况。方法:对人HCN2基因全序列进行分析,进行oligo设计,通过PCR,扩增HCN2全长cDNA,通过双酶切(XhoI和BamHI)装入真核表达载体pIRES2-EGFP中,脂质体法转染入HEK293细胞中,利用真核表达载体中带有绿色荧光蛋白GFP报告基因,对转染效率进行监测,采用反转录-聚合酶链反应检测HCN2 mRNA表达,全细胞膜片钳技术检测HCN2通道电流。结果:测序及酶切结果表明HCN2基因正确,荧光显微镜下,转染细胞观察到绿色荧光,反转录-聚合酶链反应检测到HCN2 mRNA表达,膜片钳检测到hHCN2基因编码的通道电流。结论:成功地构建了HCN2真核表达载体并进行了起搏通道HCN2基因的异源性表达。
|
| 关 键 词： | HCN4基因 真核表达载体 转染 异源性表达 |
Construction of eukaryotic expressing vector of human HCN2 geneand its expression in HEK293 |
| |
| ZUO Guang-feng,CHEN Shao-liang,XU Yan,XIAO Hang. Construction of eukaryotic expressing vector of human HCN2 geneand its expression in HEK293[J]. Progress in Modern Biomedicine, 2011, 11(6): 1068-1071 |
| |
| Authors: | ZUO Guang-feng CHEN Shao-liang XU Yan XIAO Hang |
| |
| Affiliation: | ZUO Guang-feng1,CHEN Shao-liang1,XU Yan2,XIAO Hang2(1 Nanjing First Hospital of Nanjing Medical University,Nanjing,Jiangsu 210006,China,2 Jiangsu Animal Experimental Center for Medical and Phamaceutical Research,Nanjing Medical University,Jiangsu 210029,China) |
| |
| Abstract: | Objective:To construct the eukaryon expression vector of human HCN2 gene and to investigate its expression in human embryonic kidney cells(HEK293).Methods:HCN2 gene entire sequence was analysed and designed by oligo.cDNA encoding human HCN2 gene was amplified by polymerase chain reaction,and digested by the restriction endonucleases Xho1 and BamHI,then inserted into eukaryotic expressing vector pIRES2-EGFP.pIRES2-HCN2-EGFP was transfected into HEK293 cells by Lipofacta mine2000.The transfection rate of targ... |
| |
| Keywords: | HCN2 gene Eukaryon expression vector Transfection Heterologous expression |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
| 点击此处可从《现代生物医学进展》浏览原始摘要信息 |
|
点击此处可从《现代生物医学进展》下载免费的PDF全文 |
|
