荧光蛋白mNeonGreen、mKOk作为FRET传能对在研究植物蛋白质互作中的应用分析  

Fluorescent proteins mNeonGreen and mKOk as a novel donor/acceptor pair for fluorescence resonance energy transfer in analyzing plant protein-protein interactions

作　　者：王亚琴 李宗迪 李晨羊 李云琴 胡涛 谢礼 周雪平 WANG Yaqin;LI Zongdi;LI Chenyang;LI Yunqin;HU Tao;XIE Li;ZHOU Xueping(State Key Laboratory of Rice Biology,Institute of Biotechnology,Zhejiang University,Hangzhou 310058,China;Analysis Center of Agrobiology and Environmental Sciences,Zhejiang University,Hangzhou 310058,China;Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing 100193,China)

机构地区：[1]水稻生物学国家重点实验室,浙江大学生物技术研究所,杭州310058 [2]浙江大学农生环测试中心,杭州310058 [3]中国农业科学院植物保护研究所,北京100193

出　　处：《植物保护》2021年第2期20-27,55,共9页Plant Protection

基　　金：浙江大学实验技术研究项目(SJS201809);国家自然科学基金(31972235)。

摘　　要：蛋白质相互作用是蛋白质组学研究的重要方向之一,对解析植物病原微生物与寄主植物之间的博弈具有十分重要的意义。荧光共振能量转移(fluorescence resonance energy transfer,FRET)作为一种新技术,能够在活细胞的正常生理条件下进行检测,并具有灵敏度高,假阳性较低,实验周期短的优势,在研究蛋白质相互作用过程中得到了广泛的应用。本研究选取了2种新型的荧光蛋白mNeonGreen/mKOk作为FRET传能对,并与传统的传能对CFP/YFP相比较,证实mNeonGreen/mKOk背景干扰较低。通过构建并表达荧光蛋白与目标蛋白的融合蛋白,利用FRET技术成功地在植物细胞中验证了蛋白质间的相互作用。本研究为FRET技术在植物细胞中的应用提供了更多的技术支持和借鉴。The protein-protein interaction is one of the most important fields in proteomics research,which plays an important role in understanding the molecular arms race between plant pathogens and host plants.As a new technology,fluorescence resonance energy transfer(FRET)technique has the advantages of high sensitivity,low false positive and short experiment period,and has been widely employed in detecting protein-protein interactions in living cells.In this work,two novel fluorescent proteins mNeonGreen and mKOk were used as a donor/acceptor pair for FRET assay.Compared with traditional fluorescent proteins pair CFP/YFP,mNeonGreen/mKOk pair had lower background interference.By constructing and expressing fusion proteins of fluorescent proteins and target proteins,protein-protein interactions were successfully demonstrated in plant cells by FRET.This study provides further technical support and reference for the application of FRET technique in plant cells.

关 键 词：蛋白质互作 荧光共振能量转移 mNeonGreen mKOk 

分 类 号：S432.1[农业科学—植物病理学]