摘要
研究阳离子脂质体在不同细胞中的转染效率及毒性。首先采用DNA延滞实验研究Lipofectamine2000、DOTAP转染试剂与DNA的结合能力。然后选用绿色荧光蛋白的质粒pGFP-N2作为报告基因模型和转染试剂制成复合物后转染Hela7、721、HT29细胞,比较多种因素对转染的影响。最后使用MTT比色法分析Lipofectamine2000、DOTAP转染试剂对细胞的毒性。随着复合物中转染试剂比例的增加,Lipofectamine2000、DOTAP与DNA结合能力逐渐增强。Lipofectamine2000在Hela细胞转染最高效率约72%;在7721、HT29细胞转染效率较低,DOTAP在3种细胞的转染效率都很低;在转染效率最高时2种转染试剂的细胞存活率均在90%以上。对商品化的转染试剂研究和讨论以期为阳离子类脂非病毒载体的研究提供参考。
The transfection efficiency and eytotoxicity of cationic liposome in different cells were studied. The combining ability of Lipofectamine2000, DOTAP and DNA was studied through DNA delay test. With the increasing of the proportion of transfection reagent in the complex, DNA delay was strengthened. When the mass ratio of Lipofectamine2000 to DNA and DOTAP to DNA was increased to 3 ： 1 and 6 ： 1 respectively, only a little DNA moved out from the origin site. Furthermore, pGFP-N2 that can express the green fluorescent protein was chosen as a reporter to observe the transfection efficiency directly. Then, Lipofectamine2000/DNA complexes were transfected into several cell lines, such as Hela, 7721, HT29, and the effects of the transfection conditions were evaluated. The transfection efficiency of Lipofectamine2000 to Hela cell was about 72% and higher than other 2 kinds of cell types obviously, but the transfectian efficiency of DOTAP was low. Cytotoxicity studies suggested that transfection reagents had considerably lower toxicity; the percentage of viable cells was higher than 90% in 7721 cells.
出处
《安徽农业科学》
CAS
北大核心
2009年第17期7862-7864,共3页
Journal of Anhui Agricultural Sciences
基金
国家自然科学基金资助项目(20876027)
辽宁省科技厅博士启动基金资助项目(20031082)
关键词
阳离子脂质体
转染效率
MTT
Cationic liposome
Transfection efficiency
MTT
