| grp78真核表达载体构建及其稳定转染HeLa细胞系建立 |
| |
| 引用本文： | 王恬,陶涛,杨润峰,吴章颖,李伟,廖书杰,王世宣.grp78真核表达载体构建及其稳定转染HeLa细胞系建立[J].国外医学：分子生物学分册,2011(4):309-312. |
| |
| 作者姓名： | 王恬 陶涛 杨润峰 吴章颖 李伟 廖书杰 王世宣 |
| |
| 作者单位： | 华中科技大学同济医学院附属同济医院妇产科妇科肿瘤实验室,武汉市430030 |
| |
| 基金项目： | 国家自然科学基金（No.81001152,81071663,30901586） |
| |
| 摘 要： | 目的构建人grp78基因真核表达载体，并建立稳定高表达grp78的人宫颈癌HeLa细胞系。方法用RT—PCR方法从人宫颈癌HeLa细胞中扩增grp78基因编码区，将PCR产物克隆到pcDNA3．1（＋）真核表达载体，构建重组质粒pcDNA3．1（＋）／grp78并测序鉴定。用构建成功的pcDNA3．1（＋）／grp78真核表达载体转染入人宫颈癌HeLa细胞，经G418筛选获得grp78稳定高表达的HeLa细胞系，并用RT—PCR及Western印迹方法鉴定。结果成功构建pcDNA3．1（＋）／grp78真核表达载体，筛选获得稳定高表达人grp78的HeLa细胞系。结论grp78真核表达载体的成功构建和稳定高表达grpTS的HeLa细胞系的建立为进一步研究grp78的功能奠定了基础。
|
| 关 键 词： | 人grp78基因 载体构建 转染 基因表达 |
Establishment of Eukaryotic Expression Vector of Human grp78 and Its Stable Expression in HeLa Cell Line |
| |
| Authors: | WANG Tian TAO Tao YANG Runfeng WU Zhangying LI Wei LIAO Shujie WANG Shixuan |
| |
| Institution: | Cancer Biology Research Center, Tongfi Hospital, Tongii Medical College, Huazhong University of Science ＆ Technology, Wuhan, 430030, China |
| |
| Abstract: | Objective To construct eukaryotic expression vector of human grp78 and establish its stable transfected HeLa cell line. Methods The CDS of grp78 was amplified from HeLa cell line by RT-PCR and cloned into pcDNA3.1 （ ＋ ） . The recombinant plasmid pcDNA3.1 （ ＋ ） /grp78 was sequenced. HeLa cells were transfeeted with pcDNA3.1 （ ＋ ） /grp78 and selected with G418. The expression of grp78 was identified by RT-PCR and Western Blot. Results The eukaryot- ic expression plasmid of pcDNA3.1 （ ＋ ） /grp78 was successfully constructed and grp78 stably ex- pressing HeLa cell line was established. Conclusion The recombinant eukaryotic expression vector pcDNA3.1 （ ＋ ） /grp78 and the established Hela cell line stably expressing grp78 may be used for further study of grp78 function in vitro. |
| |
| Keywords: | grp78 gene construction of vector transfection gene expression |
| 本文献已被 维普 等数据库收录！ |
