| 绿色荧光蛋白标记的葡萄糖转运蛋白4稳定表达细胞系的建立 |
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| 引用本文： | 范俊梅. 绿色荧光蛋白标记的葡萄糖转运蛋白4稳定表达细胞系的建立[J]. 现代生物医学进展, 2007, 7(10): 1467-1469 |
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| 作者姓名： | 范俊梅 |
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| 作者单位： | 华中科技大学生命科学与技术学院,湖北,武汉,430074 |
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| 基金项目： | 国家重点基础研究发展计划(973计划) |
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| 摘 要： | 目的:建立稳定表达EGFP标记的葡萄糖转运蛋白4的CHO细胞系,为研究GLUT4在CHO细胞中的转运调节机制奠定基础。方法:采用分子克隆方法构建GLUT4-EGFP的融合蛋白,在FLP-in的CHO细胞系中表达,潮霉素筛选后得到稳定的细胞系。结果:通过共聚焦显微镜的检测,证明了此稳定细胞系的阳性率达到了99%。定位研究表明大部分GLUT4以囊泡形式分布在CHO细胞胞浆内,但是质膜上也有少量的GLUT4。结论:建立了一个稳定表达GLUT4-EGFP的CHO细胞系,为进一步研究GLUT4的转运提供了一个很好的细胞模型。
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| 关 键 词： | 葡萄糖转运蛋白4 构建 CHO细胞系 |
| 文章编号： | 1673-6273（2007）10-1467-03 |
| 修稿时间： | 2007-06-28 |
Establishment of a Stable CHO Cell Line Expressing EGFP Labeled GLUT4 |
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| FAN Jun-mei. Establishment of a Stable CHO Cell Line Expressing EGFP Labeled GLUT4[J]. Progress in Modern Biomedicine, 2007, 7(10): 1467-1469 |
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| Authors: | FAN Jun-mei |
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| Affiliation: | College of Life Science and Technology, Huazhong University of Science and Technology. Wuhan, 430074, Hube, China |
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| Abstract: | Objective:To establish a stable CHO cell line expressing GLUT4-EGFP,and to lay a foundation for investigating the translocation mechanisms of GLUT4 in CHO cells.Methods:Recombinant plasmid pcDNA5/FRT-GLUT4-EGFP was constructed, co-transfected with pOG44 into FLP-in-CHO cell by lipofectamine,positive clones were screened by Hygromycin.Results:Confocal mi- croscopy image showed that over 99% CHO cells expressed GLUT4-EGFP.Further distribution results showed that most of GLUT4 was stored within vesicles in cytosol in CHO cells,but a little GLUT4 was located on the plasma membrane.Conclusion:A stable CHO cell line expressing GLUT4-EGFP was established,which provided a good cell model for studying GLUT4 function and the mechanisms of GLUT4 translocation. |
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| Keywords: | Glucose transporter4(GLUT4) Construction CHO cell line |
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