植物细胞质介导GFP-NLS蛋白入核转运的HeLa细胞系统的建立  

Establishment of a HeLa Cell System for Nuclear Translocation of Plant Cytoplasm-mediated GFP-NLS Protein

作　　者：李瑞沙 洪强[1] 周树敏[1] 张红莉[1] 李杨[1] 张卫[1] 

机构地区：[1]上海大学生命科学学院上海市能源作物重点实验室,上海200444

出　　处：《中国生物化学与分子生物学报》2017年第1期98-102,共5页Chinese Journal of Biochemistry and Molecular Biology

基　　金：上海市自然科学基金(No.15ZR1416700);国家自然科学基金(No.30870225)~~

摘　　要：细胞核作为细胞中重要的遗传物质存储、复制和转录的结构,涉及大量信息和物质的传输活动,尤其是蛋白质的入核转运一直以来都是研究的热点问题之一。本研究证明,植物细胞质可以有效应用于动物细胞体系研究蛋白质入核转运。本文利用病毒SV40抗原蛋白中的核定位信号(nuclear localization signal,NLS)标记绿色荧光蛋白(green fluorescent protein,GFP),通过拟南芥细胞质的介导,利用He La细胞核建立研究蛋白质入核转运的半细胞体系。结果显示,植物细胞质结合NLS片段能改变GFP在He La细胞核内外的分布,实现对目标蛋白入核过程的介导,使GFP-NLS最后定位于细胞核内。这也意味着通过He La细胞建立起的半细胞体系能为蛋白质入核转运研究提供一个有效的研究体系。As an important genetic material storage and the structure of replication and transcription,nucleus possesses a large amount of information and material transfer activity. Especially,the nuclear translocation of proteins has been one of the hot issues in the research. Here,we demonstrate that the plant cytoplasm can be effectively used in animal cell system to study the nuclear transfer of protein. In this paper,the nuclear localization signal（ NLS） of SV40 has been used to label GFP protein,and then by combining Arabidopsis cytoplasmic and HeLa cell nucleus,we established a semi-cell-system to study nuclear transport of the target protein. The results showed that the plant cytoplasmic binding of NLS fragments could change the distribution of GFP in the nucleus of HeLa cell,which mediate the in-nuclear transfer process of target protein, so that GFP-NLS was located in the nucleus. That means the establishment of a semi-cell-system through HeLa cells can provide an effective research system for the nuclear transport study of proteins.

关 键 词：入核转运 核定位信号 绿色荧光蛋白 细胞核 蛋白质 

分 类 号：Q71[生物学—分子生物学] Q75