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  

Establishment of a Stable CHO Cell Line Expressing EGFP Labeled GLUT4

作　　者：范俊梅[1] 

机构地区：[1]华中科技大学生命科学与技术学院,湖北武汉430074

出　　处：《现代生物医学进展》2007年第10期1467-1469,共3页Progress in Modern Biomedicine

基　　金：国家重点基础研究发展计划资助项目(2004CB720000;2006CBOD1704)

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。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 microscopy 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.

关 键 词：葡萄糖转运蛋白4 构建 CHO细胞系 

分 类 号：Q503[生物学—生物化学]