人神经母细胞瘤NRDR新剪接亚型A2的亚细胞定位  

Expression and Localization of NRDRA2 a Novel Isoform in Human Neuroblastoma

作　　者：李蕊[1] 宋旭红[1] 刘戈飞[1] 梁斌[1] 谢健平[1] 杜昆[1] 张巧霞[1] 黄东阳[1] 

机构地区：[1]汕头大学医学院分子生物学中心,广东汕头515041

出　　处：《癌变．畸变．突变》2007年第3期215-218,共4页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家自然科学基金资助(No.30470396);广东省自然科学基金博士启动项目(No.05301075)

摘　　要：背景与目的:研究hNRDRA2在真核细胞中的定位情况,并验证其羧基端预测的核定位信号(NLS)是否能引导蛋白核输入。材料与方法:分别构建hNRDRA2和NLS的绿色荧光蛋白(GFP)融合表达载体,瞬时转染人神经母细胞瘤细胞SK-N-SH、KP-N-NS及猴胚肾细胞COS-7,观察融合蛋白在3种细胞中的分布情况。结果:测序表明重组质粒构建正确,分别转染3种不同的细胞后,融合蛋白GFP-NLS表达阳性的细胞主要分布于细胞核中,GFP-A2表达阳性的细胞弥散分布于细胞质。结论:预测的NLS具有核定位功能,但携带该NLS的hNRDRA2外源表达的蛋白并不定位于细胞核。BACKGROUND ＆ AIM： To analyze the exogenous expression and localization of hNRDRA2 in eukaryocyte and demonstrate the function of the predicted nuclear localization signal （NLS） . MATERIALS AND METHODS： hNRDRA2 cDNA and NLS sequence were cloned into pEGFP-C1 to construct mammalian expression vectors pEGFP-C1-A2 and pEGFP-C1-NLS fused with green fluorescent protein （GFP） . Then these constructed vectors were transiently transfected into SK-N-SH, KP-N-NS and COS-7 cells. The transfected cells were examined under fluorescent microscope. RESULTS： The recombinant plasmids were identified by sequencing. In the transfected cells, recombinant protein GFP-A2 was distributed throughout the cell, while GFP-NLS was detected mainly in the nucleus. CONCLUSION： The predicted NLS could import the fusion protein GFP into nucleus, but exogenous expressed hNRDRA2 was not localized in the nucleus.

关 键 词：hNRDRA2 核定位信号 亚细胞定位 绿色荧光蛋白 重组表达 

分 类 号：Q786[生物学—分子生物学]