脂质体介导转染培养细胞的基因表达及转染程序的简化

A Simplified Protocol for Liposome-mediated Transfection and Gene Expression in Culture Cells

目的:以绿色荧光蛋白(green fluorescent protein,GFP)作为报道基因,观察脂质体介导转染培养细胞后基因的表达时间和表达效率,并探讨简化转染程序的影响.方法:用MCF-7细胞分别以无血清和低血清转染PEGFP-C2,从转染后4 h开始,每隔1 h在荧光显微镜下观察.结果:用无血清和低血清转染的转染效率无明显区别,5～6 h开始表达,20～24 h达到最强,48 h荧光未见减弱,72 h仍可见荧光.结论:血清不影响转染效率,能简化转染程序.基因在转染5 h后开始表达,暗示转染10 h后,基因表达可能属于其相关基因表达产物对细胞生理活动的刺激影响.

Objective：Using GFP as reporter,to determine the gene expression time, expression efficiency of the fiposome-mediatedly transfected cells in a simplified protocol, Method： The MCF-7 cells was transfected with PEGFP- C2 by serum-free or serum medium, The cell emitting fluorescence and the intensity were observed under microscope at every one-hour interval beginning from 4 hrs post-transtection, Result： There is no obvious transfection efficiency difference between serum-free and low serum medium groups. The fluorescence occurred at 5 - 6 hours after transfection, reached maximum at 20 - 24 hours, maintained relative intensity at 48 hours, and declined at 72 hours, Conclusion： The serum does not affect efficiency. This may be used to design simplifed procedure of experiment. Transfection program can be simplified with the presence of serum. The gene introduced into the cells can express itself as early as 5 hour after transfection, suggesting that the index relevant to the gene production after 10 hours of transfection may be the results of indirect impact.