| 稳定表达绿色荧光蛋白的COS7细胞系的筛选和鉴定 |
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| 引用本文： | 龙鼎新,李小玲,温元武※.稳定表达绿色荧光蛋白的COS7细胞系的筛选和鉴定[J].南华大学学报(医学版),2012,40(1):11-13. |
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| 作者姓名： | 龙鼎新 李小玲 温元武※ |
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| 作者单位： | 南华大学公共卫生学院,湖南衡阳,421001 |
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| 基金项目： | 国家自然科学基金，湖南省自然科学基金，湖南省教育厅优秀青年基金，南华大学博士启动基金 |
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| 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500 μg/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.
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| 关 键 词： | COS7 GFP 转染 G418筛选 细胞克隆 |
| 收稿时间： | 2011/10/21 0:00:00 |
Establishment and Identification of Stable Expressing Green Fluorescence Protein COS-7 Cell Line |
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| LONG Ding-xin,LI Xiao-ling,WEN Yuan-wu.Establishment and Identification of Stable Expressing Green Fluorescence Protein COS-7 Cell Line[J].Journal of Nanhua University(Medical Edition),2012,40(1):11-13. |
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| Authors: | LONG Ding-xin LI Xiao-ling WEN Yuan-wu |
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| Institution: | School of Public Health,University of South China,Hengyang,Hunan 421001,China |
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| Abstract: | ObjectiveTo establish a stable expressing green fluorescence protein (GFP) COS-7 cell line and to provide a convenient control for some experiments.MethodCOS-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 |
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| Keywords: | COS-7 GFP transfection G418 screening cell cloning |
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