双分子荧光互补技术及其在蛋白互作研究中的应用

Biomolecular Fluorescence Complementation(BiFC) and Its Application in Researching on Protein-Protein Interactions

双分子荧光互补(bimolecular fluorescence complementation,BiFC)是近年新发展起来的在活细胞中研究蛋白质互作及其定位的新技术.该技术巧妙的将荧光蛋白切成不发光的两个片段,并与目的蛋白融合表达.如果目的蛋白相互作用,连接其上的荧光互补蛋白片段将会相互靠近并恢复荧光活性,从而实现了分子与细胞事件的可视化.BiFC既可以验证蛋白之间的互作、检测其互作强弱,还可以定位互作蛋白的位点.基于双分子荧光蛋白的应用已经越来越广泛,本文对双分子荧光互补技术的原理、应用现状、特点及其面临的问题进行了概述.

Biomolecular fluorescence complementation is a new technique that developed inrecent years to research on protein-protein interactions and their subcellular location.It delicately seprated fluorescence protein into two non-fluorescent fragments.If the interest proteins interact with each other, on which the fragments that fused respectively will be adjacent and recovery fluorescence property,and therefore come true visualization of molecular and cell events.BiFC can both verificate the protein-protein interation and their intensity and locate their interaction sites.Applications based on BiFC have became more and more extensive.The text descripted BiFC＇ s developmnent, principle ,characters ,current situation of its application and the problems it faced with.