利用慢病毒载体快速建立表达外源基因的哺乳动物细胞系  

Rapid Generation of a Stable Mammalian Cell Line Expressing Exogenous Gene Using Lentiviral Vector

作　　者：陈平[1,2] 胡涛[2] 薛敬礼[2] 杜春燕[2] 章金涛[2] 朱奎成[2] 金树兴[2] 王纯耀[2] 

机构地区：[1]郑州大学基础医学院病理生理教研室,郑州450001 [2]郑州大学实验动物中心,郑州450001

出　　处：《生物技术通报》2011年第7期210-213,共4页Biotechnology Bulletin

基　　金：国家自然科学基金青年基金项目(81001102)

摘　　要：建立稳定表达外源基因的哺乳动物细胞系是一项重要的生物技术,介绍了联合使用慢病毒载体和流式细胞仪分选技术来建立稳定表达绿色荧光蛋白的293T细胞系。结果表明,在1个月左右的时间里即可获得表达绿色荧光蛋白的293T细胞系,阳性细胞率高达96.5%,而且建立的细胞系能够稳定传代。因此,联合慢病毒载体和流式细胞仪分选技术的策略对于建立稳定表达外源基因的哺乳动物细胞系快捷和可靠。The generation of stable mammalian cell lines expressing exogenous gene is an important technique in cell research,gene therapy and protein production.An integrated strategy based on lentiviral vector and fluorescence activated cell sorting（FACS）to facilitate this process was described.Using integrated strategy,we can establish a 293T cell line expressing enhanced green fluorescent protein（EGFP）in about one month.The number of EGFP-positive cells was more than 96.5% which were stable during passaging.Therefore,the strategy described here is fast and reliable for the generation of mammalian cell lines expressing exogenous gene.

关 键 词：慢病毒载体 流式细胞仪分选技术 绿色荧光蛋白 哺乳动物细胞系 

分 类 号：Q78[生物学—分子生物学]