| 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].中南医学科学杂志.,2012,40(1):11-13. |
| 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 |
| 投稿时间：2011-10-21 |
| DOI： |
| 中文关键词: COS7 GFP 转染 G418筛选 细胞克隆 |
| 英文关键词:COS-7 GFP transfection G418 screening cell cloning |
| 基金项目: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） |
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| 中文摘要: |
| 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。 |
| 英文摘要: |
| 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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