【摘要】 目的探究采用电穿孔法将质粒pGenesil-1(简称p G)转染老年人成纤维细胞,从中寻找最佳的参数。方法采用电穿孔法将能表达G418抗性蛋白和增强型绿色荧光蛋白(EGFP)的质粒p G转染老年人成纤维细胞。荧光显微镜下观察EGFP表达并测定转染效率、细胞存活率;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老年人成纤维细胞转染的最佳电转条件:电压:240 V;电容:750μF;DNA剂量:8μg。48 h后电穿孔法的转染率接近30%。结论用电穿孔法将质粒pG稳定转染老年人成纤维细胞耗时短,有利于在体外对老年人成纤维细胞进行后续实验。更多还原

【Abstract】 Objective To transfect plasmid p Genesil-1( p G) into elderly so as to fibroblasts find the best parameters. Methods Electroporation was used for transfect plasmid p G,a plasmid that expresses G418 resistance protein and enhanced green fuluorescent protein( EGFP),into elderly fibroblasts. The expression of EGFP was observed and the 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 elderly fibroblasts transfection were: power transfer voltage of240 V,capacitance of 750 μF,DNA dose of 8 μg. Transfection efficiency of electroporation after 48 h was about 30%. Conclusion Stably transfecting plasmid p G into elderly fibroblasts with electroporation need short time,and this will help to take advantage of the elderly fibroblasts for subsequent experiments.更多还原