首页 | 本学科首页   官方微博 | 高级检索  
     

聚乙烯亚胺介导水通道蛋白5基因表达载体体外转染的研究
引用本文:刘洋,王月兰,宋秀梅. 聚乙烯亚胺介导水通道蛋白5基因表达载体体外转染的研究[J]. 山东大学学报(医学版), 2011, 49(8): 57-61
作者姓名:刘洋  王月兰  宋秀梅
作者单位:山东大学附属千佛山医院麻醉科, 济南 250014
摘    要:目的 探讨新型阳离子载体聚乙烯亚胺(PEI)介导水通道蛋白5(AQP5)体外转染人肺腺癌细胞系A549的效果。方法 将含绿色荧光蛋白的真核表达载体pEGFP-N1,以lipofectamine2000为阳性对照,流式细胞仪检测转染效率,根据不同N/P比(5、10、15、20、25)与PEI组成转染复合物转染A549细胞,优化最适 N/P比;将AQP5全长cDNA插入pEGFP-N1载体,构建pEGFP-N-AQP5,转染A549细胞,采用PCR与Western blot法检测被转染细胞中AQP5的表达。结果 流式细胞仪检测GFP发光率,N/P比为15时PEI/pEGFP-N1复合物转染效率最高,可达到89.72%,与脂质体91.44%相近;应用PEI与lipofectamine2000将 pEGFP-N-AQP5成功转染A549,与空白组比较,AQP5的基因及蛋白表达均增高。结论 新型阳离子载体PEI 可成功介导pEGFP-N-AQP5转染A549细胞。

关 键 词:聚乙烯亚胺;转染;水通道蛋白  
收稿时间:2010-10-08

Transfection of the AQP5 gene mediated by a novel cational carrier PEI in vitro
LIU Yang,WANG Yue-lan,SONG Xiu-mei. Transfection of the AQP5 gene mediated by a novel cational carrier PEI in vitro[J]. Journal of Shandong University:Health Sciences, 2011, 49(8): 57-61
Authors:LIU Yang  WANG Yue-lan  SONG Xiu-mei
Affiliation:Department of Anesthesiology, Qianfoshan Hospital Affiliated to Shandong University, Jinan 250014, China
Abstract:Objective To investigate the effect of a new cationic carrier polyethylenimine (PEI) on transfection of the aquaporin 5 (AQP5) gene into A549 cells in vitro. Methods According to different N/P ratios (5, 10, 15, 20 and 25), the eukaryotic expression vector pEGFP-N1 was mixed with the green fluorescent protein(GFP) and PEI to make a transfection complex, using lipofectamine2000 as a positive control. Transfection efficiency was detected by flow cytometry for optimizing the optimal N/P ratio. Full length AQP5 cDNA were cloned into the pEGFP-N1 vector to construct pEGFP-N-AQP5, which was then transfected into A549 cells. Expression of AQP5 in transfected cells was detected using PCR and Western blot. Results Flow cytometry revealed the GFP emitting. When the N/P ratio was 15, the PEI/pEGFP-N1 complex achieved the highest transfection efficiency of 89.72%, close to 91.44% when delivered by liposomes. Compared with the control group, AQP5 gene and protein expressions were significantly increased after successful transfection of pEGFP-N-AQP5 into A549 by application of the PEI and lipofectamine2000 complex. Conclusion pEGFP-N-AQP5 can be successfully transfected into A549 cells mediated by the novel cationic carrier PEI.
Keywords:Polyethylenimine   Transfection   Aquaprins,
本文献已被 CNKI 万方数据 等数据库收录!
点击此处可从《山东大学学报(医学版)》浏览原始摘要信息
点击此处可从《山东大学学报(医学版)》下载全文
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号