copine Ⅴ蛋白的亚细胞定位  

Subcellular Localization of copine Ⅴ

作　　者：王晓文[1] 靳雁斌[1] 吴燕[1] 吴彦瑞[1] 马鑫[1] 刘淑红[1] 范文红[1] 范明[1] 

机构地区：[1]军事医学科学院基础医学研究所,北京100850

出　　处：《生物技术通讯》2006年第2期149-151,共3页Letters in Biotechnology

基　　金：国家高技术研究发展计划课题(2002BA711A01-03)

摘　　要：目的:确定copineⅤ蛋白的亚细胞定位,初步研究该蛋白的生物学功能。方法:将copineⅤ编码区基因分别构建真核表达载体pEGFP-copineⅤ(或pRED-copineⅤ),转染HEK293、HeLa细胞,在激光共聚焦荧光显微镜下与转染空载体pEGFP-N1(或pRED-N1)的细胞比较观察。结果:经限制性内切酶分析鉴定,构建的重组表达载体正确。通过激光共聚焦荧光显微镜观察,转染了重组载体pEGFP-copineⅤ的细胞荧光信号集中分布于胞膜和内膜系统;进一步研究表明copineⅤ定位于内质网而非线粒体,而空载体则在整个细胞中均匀分布。结论:copineⅤ蛋白定位于细胞膜和内质网上,而不定位于线粒体。Objective： To investigate the subcellular localization of copine V gene. Methods： The expressive vector of pEGFP-copineV （or pRED-copineV ） was constructed by routine molecular biological, HEK293 and HeLa cells were transfected with pEGFP-N1 and pEGFP-copineV, respectively by using LipofectAMINE2000. The expression was dectected by fluorescence microscope and confocal fluorescence microscope. Results： The expression vectors were confirmed by re- strietion enzyme disgestion. The green and red fluorescence could be seen in cell membrane and endoplasmic reticulum rather than mitochondrial in transfected copine V gene cells and were distributed uniformly in whole cell in transfected pEGFP-N1 （or pRED-N1 ） cells respectively. Conclusion： The expression of copineV gene was distributed in cell membrane and endoplasmic reticulum rather than mitochondrial.

关 键 词：copine V蛋白 亚细胞定位 内质网 线粒体 细胞膜 

分 类 号：Q25[生物学—细胞生物学]