稳定表达人Eps8基因的胶质瘤U251细胞系的建立  

Establishment of U251 Cell Line for Stable Expression of Human Eps8 Gene

作　　者：周芳亮[1] 胡翔[1] 杨子剑[1] 韩梅[1] 丁小凤[1] 

机构地区：[1]湖南师范大学生命科学学院蛋白质化学与发育生物学教育部重点实验室,湖南长沙410081

出　　处：《激光生物学报》2010年第4期449-452,共4页Acta Laser Biology Sinica

基　　金：国家自然科学基金项目(30900827)

摘　　要：应用亚克隆方法构建pEGFP-C3/Eps8真核表达载体,经测序鉴定后,用脂质体进行胶质瘤U251细胞的转染,应用G418筛选出稳定表达pEGFP-C3/Eps8和pEGFP-C3的细胞系,最后通过Western blot和荧光定位证明Eps8在U251细胞中过量表达。本实验成功建立了稳定转染Eps8的U251细胞系,为进一步研究Eps8基因在胶质瘤中的功能奠定了良好的实验基础。The eukaryotic expression vector pEGFP-C3/Eps8 was constructed by the sub-cloning. After verified by sequencing,we transfected the plasmids into U251 cells by LipofectamineTM 2000. After screening by G418, stable transfected cell lines were established to express pEGFP-C3/Eps8 and pEGFP-C3. Finally, the overexpression of Eps8 was demonstrated by western blotting and fluorescence localization assays. Taken together, U251 cell line stably expressing Eps8 is established, which will lay a solid experimental foundation for further studies on the function of Eps8 gene in glioma cells.

关 键 词：印娼基因 真核表达载体 U251细胞 稳定表达 

分 类 号：Q78[生物学—分子生物学]