稳定表达绿色荧光蛋白的COS7细胞系的筛选和鉴定  

Establishment and Identification of Stable Expressing Green Fluorescence Protein COS-7 Cell Line

作　　者：龙鼎新[1] 李小玲[1] 温元武[1] 

机构地区：[1]南华大学公共卫生学院,湖南衡阳421001

出　　处：《中南医学科学杂志》2012年第1期11-13,50,共4页Medical Science Journal of Central South China

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

摘　　要：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 COS7-GFP cells maintain stable biology character and display bright green fluorescence under fluorescent microscope,and GFP protein produced in these cells can be detected by western blotting.Conclusion Stable GFP-expression COS-7 cell line（COS7-GFP cell line） was established successfully.This cell line can be handily used as a control for some experiments.

关 键 词：COS7 GFP 转染 G418筛选 细胞克隆 

分 类 号：Q813[生物学—生物工程]