Fan Jin-Yu, Cui Zong-Qiang, Wei Hong-Ping, Zhang Zhi-Ping, Zhou Ya-Feng, Wang Yun-Peng, Zhang Xian-En
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, 44#, Xiao Hong Shan, Wuhan 430071, China.
Biochem Biophys Res Commun. 2008 Feb 29;367(1):47-53. doi: 10.1016/j.bbrc.2007.12.101. Epub 2007 Dec 26.
Bimolecular fluorescence complementation (BiFC) is a recently developed technique for detection of protein-protein interactions in living cells. In this study, a new red BiFC system was developed by splitting mCherry, a mutant monomeric red fluorescent protein, into two fragments between amino acids 159-160 and was verified using a pair of interacting proteins, SV40 large T antigen (LTag), and human p53 protein. By combined use of the mCherry-based red BiFC system with a Venus-based yellow BiFC system, the interaction between LTag and p53 as well as the interaction between sp100 and promyelocytic leukemia protein (PML), were detected simultaneously in Vero cells. The brilliant redness, short maturation time, and the long excitation and emission wavelengths (587/610 nm) of mCherry make the new BiFC system an excellent candidate for analyzing protein-protein interactions in living cells and for studying multiple protein-protein interactions when coupled with other BiFC systems.
双分子荧光互补（BiFC）是一种最近开发的用于检测活细胞中蛋白质-蛋白质相互作用的技术。在本研究中，通过将突变型单体红色荧光蛋白mCherry在氨基酸159 - 160之间拆分成两个片段，开发了一种新的红色BiFC系统，并使用一对相互作用的蛋白质，即SV40大T抗原（LTag）和人p53蛋白进行了验证。通过将基于mCherry的红色BiFC系统与基于Venus的黄色BiFC系统联合使用，在Vero细胞中同时检测到了LTag与p53之间的相互作用以及sp100与早幼粒细胞白血病蛋白（PML）之间的相互作用。mCherry的鲜艳红色、较短的成熟时间以及较长的激发和发射波长（587/610 nm）使得新的BiFC系统成为分析活细胞中蛋白质-蛋白质相互作用以及与其他BiFC系统结合研究多种蛋白质-蛋白质相互作用的极佳候选者。
