| 人ob基因克隆及其亚细胞定位的研究 |
| |
| 引用本文： | 陈香凝,阴彦辉,施青青,孙敏,傅德智,高波,李碧春. 人ob基因克隆及其亚细胞定位的研究[J]. 生物技术, 2011, 21(2). DOI: 10.3969/j.issn.1004-311X.2011.02.035 |
| |
| 作者姓名： | 陈香凝 阴彦辉 施青青 孙敏 傅德智 高波 李碧春 |
| |
| 作者单位： | 扬州大学动物科学与技术学院,江苏扬州,225009 |
| |
| 基金项目： | 国家863高技术研究发展计划项目,江苏省高校自然科学重大基础研究项目 |
| |
| 摘 要： | 目的:旨在克隆人肥胖(obese,ob)基因的全长cDNA序列,与EGFP重组构建融合蛋白表达载体,并分析其亚细胞水平的定位.方法:提取人脂肪细胞总RNA,采用RT-PCR方法扩增出人ob基因cDNA,并克隆至真核表达载体pEGFP-CI,重组质粒转染NIH-3T3细胞,荧光显微镜分析EGFP-ob融合蛋白的亚细胞定位.结果:克隆的ob基因cDNA为501bp,共编码167个氨基酸,与GenBank公布的人ob基因序列一致,荧光显微镜分析表明,重组的EGFP-ob融合蛋白主要分布于NIT-3T3的细胞质中.结论:成功克隆了人OB基因的cDNA序列,构建人OB基因的真核表达载体pEGFP-CI-ob,融合蛋白EGFP-ob定位于NIH-3T3细胞质中.
|
| 关 键 词： | 人 肥胖基因 EGFP蛋白 亚细胞定位 |
Cloning and Sub-cellular Localization of Human Obese Gene |
| |
| CHEN Xiang-ning,YIN Yan-hui,SHI Qing-qing,SUN Min,FU De-zhi,GAO Bo,LI Bi-chun. Cloning and Sub-cellular Localization of Human Obese Gene[J]. Biotechnology, 2011, 21(2). DOI: 10.3969/j.issn.1004-311X.2011.02.035 |
| |
| Authors: | CHEN Xiang-ning YIN Yan-hui SHI Qing-qing SUN Min FU De-zhi GAO Bo LI Bi-chun |
| |
| Affiliation: | CHEN Xiang-ning,YIN Yan-hui,SHI Qing-qing,SUN Min,FU De-zhi,GAO Bo,LI Bi-chun(College of Animal Science and Technology,Yangzhou University,Yangzhou 225009,China) |
| |
| Abstract: | Objective:The aim of this study is to clone full-length cDNA of obese(ob) gene in humans and analysis its sub-cellular localization through EGFP fusion protein.Method:cDNA of ob gene was cloned by RT-PCR from total RNA extracted from human fat tissue.The RT-PCR product was sub cloned into pMD19-T vector.The sequence analysis was then performed.The cDNA of ob gene was inserted into upstream of an expression vector pEGFP-C1,which contains the enhanced green fluorescent protein(EGFP),to construct fusion expres... |
| |
| Keywords: | human Obese gene EGFP protein sub-cellular localization |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
|
