| 稳定表达人Eps8基因的胶质瘤U251细胞系的建立 |
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| 引用本文： | 周芳亮,胡翔,杨子剑,韩梅,丁小凤.稳定表达人Eps8基因的胶质瘤U251细胞系的建立[J].激光生物学报,2010,19(4):449-452. |
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| 作者姓名： | 周芳亮 胡翔 杨子剑 韩梅 丁小凤 |
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| 作者单位： | 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 |
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| 基金项目： | 国家自然科学基金项目 |
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| 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和荧光定位证明印娼在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。
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| 关 键 词： | 印娼基因 真核表达载体 U251细胞 稳定表达 |
Establishment of U251 Cell Line for Stable Expression of Human Eps8 Gene |
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| ZHOU Fang-liang,HU Xiang,YANG Zi-jian,HAN Mei,DING Xiao-feng.Establishment of U251 Cell Line for Stable Expression of Human Eps8 Gene[J].ACTA Laser Biology Sinica,2010,19(4):449-452. |
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| Authors: | ZHOU Fang-liang HU Xiang YANG Zi-jian HAN Mei DING Xiao-feng |
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| Institution: | (Key Laboratory of Protein Chemistry and Development Biology of State, Education Ministry of China, College of Life Science, Hunan Normal University, Changsha 410081, Hunan, China) |
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| Abstract: | 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. |
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| Keywords: | Eps8 gene mammalian expression plasmid U251 cell stable expression |
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