小鼠胎儿成纤维细胞及胚胎干细胞转绿色荧光蛋白基因的研究  

Study of transfection of enhanced green fluorescent protein gene into mouse embryonic fibroblast and embryonic stem cells

作　　者：阎晓飞[1] 徐燕宁[1] 申景岭[1] 单智焱[1] 关娜[1] 钟淑琦[1] 金连弘[1] 雷蕾[1] 

机构地区：[1]哈尔滨医科大学组织学与胚胎学教研室,黑龙江哈尔滨150081

出　　处：《哈尔滨医科大学学报》2008年第6期549-552,共4页Journal of Harbin Medical University

基　　金：国家自然科学基金资助项目(30671025);黑龙江省留学归国人员科学技术专项资金(LC07C17);哈尔滨医科大学科研启动基金

摘　　要：目的检测用增强型绿色荧光蛋白基因(EGFP)转染的小鼠胎儿成纤维细胞及小鼠胚胎干细胞的转染效率及EGFP的表达情况。方法电穿孔法将EGFP转入胎儿成纤维细胞及胚胎干细胞,G418筛选,荧光显微镜观察转染效率及绿色荧光蛋白表达情况。结果胚胎干细胞的转染率高于成纤维细胞,转染后的两种细胞经药物筛选均有绿色荧光蛋白表达。结论电穿孔转染细胞的方法简单、效率高,是一种较为理想的基因转染方法,经药物筛选后的两种细胞均可用于核移植,为转基因克隆动物的研究奠定了基础。Objective To evaluate transfection efficiency of mouse embryonic fibroblast and embryonic stem cells transfected with enhanced green fluorescent protein gene（EGFP） ,and the expression of EGFP in these cells. Methods EGFP was introduced into mouse embryonic fibroblast and embryonic stem ceils by electroporation. Transfected cells were selected by G418 resistance and screened by EGFP expression under the fluorescent microscope. Results Fluorescent proteins were expressed both in mouse embryonic fibroblast and embryonic stein cells. However, the transfection efficiency of embryonic stem cells was significantly higher than that of fibroblast. Conclusion For mouse embryonic fibroblast and embryonic stem cells, eleetroporation is a simple and efficient gene transfection method. Transfected cells which screened by drug resistance can be used as donor cells for nuclear transfer.

关 键 词：电穿孔 转基因 成纤维细胞 胚胎干细胞 

分 类 号：R321[医药卫生—人体解剖和组织胚胎学]