一种检测蛋白质相互作用的双荧光共定位系统

周 军;周翠红;邓小燕;陶凤杰

doi:10.3981/j.issn.1000-7857.2011.08.001

科技导报 >

2011 , Vol. 29 >Issue 11-08: 22 - 26

DOI: https://doi.org/10.3981/j.issn.1000-7857.2011.08.001

研究论文

一种检测蛋白质相互作用的双荧光共定位系统

周军;周翠红;邓小燕;陶凤杰

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1. 北京航空航天大学生物与医学工程学院；生物力学与力生物学教育部重点实验室，北京 1001912. 北京大学化学与分子工程学院，北京核磁共振中心，北京 1008713. 北京市第五十六中学生物教研组，北京 100044

收稿日期: 2010-12-28

修回日期: 2011-02-23

网络出版日期: 2011-03-18

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A Translocation Fluorescence Cross-correlation Tool for the Analysis of Protein-protein Interactions

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Received date: 2010-12-28

Revised date: 2011-02-23

Online published: 2011-03-18

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摘要

自发荧光蛋白以快捷灵敏、即时检测和无需破坏活细胞的特点在蛋白质相互作用的研究中得到广泛应用。从发光水母Aquoria victoria中分离的绿色荧光蛋白（GFP）和从珊瑚Discosoma sp.中分离的DsRed红色荧光蛋白是自发荧光蛋白的典型代表。本研究开发一种基于GFP和红色荧光蛋白（RFP），并可通过待检测蛋白在细胞内表达的空间位移变化产生共定位的双荧光共定位检测系统。该系统为克服两种荧光蛋白光谱渗漏带来的非特异性结果的干扰，将核仁定位信号（NoLS）和出核信号（ABL）分别连接到RFP和GFP。具有定位信号的GFP和RFP的待检测蛋白对可分别在细胞核内和（或）细胞核外表达，伴随它们的相互作用，荧光共定位的现象会产生在细胞内特定的区域。利用抗凋亡蛋白Bcl-2和Bak-BH3短肽作为蛋白对测试该系统，荧光共定位现象明显，系统灵敏可靠。

关键词： GFP; DsRed; 蛋白质相互作用

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周军;周翠红;邓小燕;陶凤杰 . 一种检测蛋白质相互作用的双荧光共定位系统[J]. 科技导报, 2011 , 29(11-08) : 22 -26 . DOI: 10.3981/j.issn.1000-7857.2011.08.001

Abstract

Fluorescence proteins have become widely used molecular probes in the analysis of protein-protein interactions. There are many notable advantages, such as celerity, direct visualization, and noninvasive. Green Fluorescence Protein (GFP) from jelly fish and Red Fluorescence Protein (RFP) from coral are most important auto-fluorescent proteins in this field. A translocation fluorescence cross-correlation tool is developed base on the GFP and RFP for testing protein-protein interaction. To overcome the distract caused by non-specific spectrum permeation of two fluorescent proteins, the nucleolus localization signal sequence (NoLS) and nuclear export signal sequence (ABL signal sequences) are attached to both pairs of the system accordingly. Therefore, the proteins can express in and/or out of the nuclear which endues locomotion of protein pairs and their fluorescence tags. The dynamic protein-protein interaction of anti-apoptotic protein Bcl-2 and Bak-BH3 peptide is straightly and rapidly laid out in the translocation fluorescence cross-correlation tool.

Key words： GFP; DsRed; protein-protein interaction

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