| 大鼠Pael -R基因shRNA干扰载体的构建及功能筛选鉴定 |
| |
| 引用本文： | 邹婷,肖波,汤建光,张贺.大鼠Pael -R基因shRNA干扰载体的构建及功能筛选鉴定[J].生物技术,2011,21(4). |
| |
| 作者姓名： | 邹婷 肖波 汤建光 张贺 |
| |
| 作者单位： | 1. 中南大学湘雅二医院神经内科,湖南长沙,410011
2. 中南大学湘雅医院神经内科,湖南长沙,410008 |
| |
| 摘 要： | 目的:构建并筛选针对大鼠Pael -R基因的有效shRNA干扰载体并鉴定其干扰效果.方法:构建三个针对大鼠Pael-R基因的shRNA表达载体,利用脂质体转染大鼠肾上腺嗜铬细胞瘤细胞PC12,以G418筛选抗性细胞克隆并利用RT-PCR和Western Blotting鉴定Pael -R基因的表达.结果:在构建的3个干扰载体中,转染pRNA-U6/PaelR -3的细胞克隆中Pael -R表达在mRNA水平为29％,与对照组相比下降了45.3％,在蛋白质水平为32％,下降了27.3％ (P＜0.05).结论:构建了Pael -R基因的有效干扰载体pRNA-U6/PaelR -3,并得到了Pael -R基因表达下调型PC12细胞克隆,为研究Pael -R基因表达下调在帕金森病的发病机制及其治疗中的作用奠定了基础.
|
| 关 键 词： | RNA干扰 Pael -R 帕金森病 |
Construction of shRNA Interference Vector for Rat Pael- R Gene and Functional Identification |
| |
| ZOU Ting,XIAO Bo,TANG Jian-guang,ZHANG He.Construction of shRNA Interference Vector for Rat Pael- R Gene and Functional Identification[J].Biotechnology,2011,21(4). |
| |
| Authors: | ZOU Ting XIAO Bo TANG Jian-guang ZHANG He |
| |
| Institution: | ZOU Ting1,XIAO Bo2,TANG Jian-guang1,ZHANG He1(1.Department of Neurology,Second Xiangya Hospital of Central South University,Changsha 410011,China,2.Department of Neurology,Xiangya Hospital of Central South University,Changsha 410008,China) |
| |
| Abstract: | Objective:To construct and select the effective shRNA interference vector for rat Pael-R gene,and to identify its effect of interference.Method: Three recombination shRNA vectors were constructed.The vectors were transfected into rat pheochromocytoma cells PC12 by lipofection 2000 and cell clones were selected by G418.The expression of Pael-R was detected by RT-PCR and Western Blotting.Result:In the cell clone transfected by pRNA-U6/PaelR-3 vector,the expression of Pael-R was 29%,decreased by 45.3% at trans... |
| |
| Keywords: | RNA interference Pael-R Parkinson's disease |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
|
