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.