期刊论文

中文

中南医学科学杂志

2095-1116

2012

40

1

11-13,50

10.15972/j.cnki.43-1509/r.2012.01.002

81172712:国家自然科学基金 11JJ6078:湖南省自然科学基金 09B087:湖南省教育厅优秀青年项目 2010XQD19:南华大学博士启动基金

本校为第一机构

目的构建稳定表达绿色荧光蛋白（green fluorescence protein,GFP）的COS-7细胞系,为相关实验对照提供方便。方法常规方法培养COS-7细胞,将含pEGFP-N3基因的质粒转染COS-7细胞,G418抗性筛选,并用荧光显微镜进行跟踪检测,挑选耐药单细胞克隆并扩大培养至稳定细胞株。结果 G418筛选的最适浓度为500μg/mL。筛选到的COS7-GFP克隆荧光显微镜下发出明亮的绿色荧光,细胞的生物学性状保持稳定,通过Western blot能检测到GFP蛋白。结论成功筛选并建立了稳定表达的COS7-GFP细胞系。

Objective To establish a stable expressing green fluorescence protein（GFP） COS-7 cell line and to provide a convenient control for some experiments.Method COS-7 cells were cultured by the conventional method.The pEGFP-N3 plasmid was transfected into COS-7 cells,and cells were screened by G418 until the stable G418-resistant monoclonal cells（COS7-GFP cells） were acquired.The GFP-expression cells were tracked by fluorescent microscope.Results The optimum G418 concentration for screening drug-resistant cell line is 500 μg/mL.All the selected ...