Analysis of nuclear localization signals using a green fluorescent protein-fusion protein library

作     者：Fujii, G Tsuchiya, R Ezoe, E Hirohashi, S 

作者机构：Natl Canc Ctr Res Inst Div Pathol Chuo Ku Tokyo 1040045 Japan JST ERATO Hirohashi Cell Configurat Project Ibaraki Osaka 3002635 Japan 

出 版 物：《EXPERIMENTAL CELL RESEARCH》 (实验细胞研究)

年 卷 期：1999年第251卷第2期

页      面：299-306页

核心收录：

学科分类：0710[理学-生物学] 1002[医学-临床医学] 07[理学] 09[农学] 

主　　题：氨基酸序列 细胞系 克隆 分子/方法 基因文库 遗传载体 绿色荧光蛋白质类 肝/细胞学 发光蛋白质类/遗传学 分子序列数据 核定位信号 核蛋白质类/遗传学 核蛋白质类/分离和提纯 肽类 重组融合蛋白质类/分离和提纯 选择(遗传学) β半乳糖苷酶类/遗传学 人类 

摘      要：We describe here an efficient method for identifying intracellular localization signals in proteins with stereospecific intracellular localizations in culture cells. The method involves rapid fluorescence screening of cells transfected with a cDNA library in which cDNAs are fused to the gene encoding the Aequorea victoria green fluorescent protein (GFP). We analyzed nuclear localization and nuclear localization signals (NLSs) in a model application of this method. As a result, we identified classical MLSs in 75% of nuclear localized proteins. We identified some novel NLS candidates among the classical NLS-negative sequences whose nuclear localization was also identified in another cell line and with other molecular tag sequences. This method will be useful for identifying intracellular localization signals and for more detailed analysis of intracellular architecture. (C) 1999 Academic Press.