【摘要】 目的:采用电穿孔法将质粒pGenesil-1(简称pG)转染32DP210细胞,从中寻找最佳的参数。方法:分别采用LipofectamineTM2000、transfast、脂质体DMRIC-E和电穿孔法,将能表达G418抗性蛋白和绿色荧光蛋白(GFP)的质粒pG转染32DP210细胞;荧光显微镜下观察GFP表达并测定瞬时转染率;通过G418筛选得到稳定转染的细胞。结果:本实验室电穿孔法介导的32DP210细胞转染其最佳电转条件为电压270 V,电容950μF,DNA剂量14μg。48 h后电穿孔法的转染效率达50.81%,其它三种方法转染效率都小于3%。结论:用电穿孔法将质粒pG转染32DP210细胞是最佳的转染方法。更多还原

【Abstract】 Objective:To find the best parameters to transfect plasmid pG into 32DP210 cells.Methods:LipofectamineTM2000,transfast,liposome DMRIC-E transfection and electroporation were compared for transfecting plasmid pG,a plasmid that expressed G418 resistance protein and green fluorescent protein(GFP),into 32DP210 cells.The expression of GFP was observed and the transient transfection efficiency was measured after the transfection by fluorescence microscope.The stable transfected cells were screened by G418.Results:The optimum conditions for laboratory electroporation-mediated 32DP210 cell transfection were:power transfer voltage 270 V,capacitance of 950 μF,DNA dose of 14 μg.Transfection efficiency of electroporation after 48 h was 50.81%,while the other three methods were less than 3%.Conclusion:Electroporation is the best way to stably transfect plasmid pG into 32DP210 cells.更多还原