Open AccessNew Fast BiFC Plasmid Assay System for <i>in Vivo</i> Protein-Protein Interactions
Author(s) -
MyungHwa Kim,
Hyo Jin Roh,
Min-Nyung Lee,
ManWook Hur
Publication year - 2007
Publication title -
cellular physiology and biochemistry
Language(s) - Uncategorized
Resource type - Journals
SCImago Journal Rank - 1.486
H-Index - 87
eISSN - 1421-9778
pISSN - 1015-8987
DOI - 10.1159/000110431
Subject(s) - bimolecular fluorescence complementation , in vivo , protein–protein interaction , protein fragment complementation assay , yellow fluorescent protein , microbiology and biotechnology , computational biology , green fluorescent protein , plasmid , biology , protein domain , complementation , cloning (programming) , fluorescent protein , chemistry , genetics , gene , computer science , programming language , phenotype
In this age of massive genetic and protein information, a fast and reliable method of studying in vivo protein-protein interactions is necessary. We have developed a novel system that can overcome limitations of existing assay methods. This new method adopts two existing systems for fast analysis of diverse protein-protein interactions. For rapid, large-scale cloning, we adopted the Gateway system and developed novel destination vectors containing YFP N-terminus (YN) or YFP C-terminus (YC) to visualize protein-protein interactions in vivo using bimolecular fluorescence complementation (BiFC). Using this system, we investigated molecular interactions among the three POZ-domain regulatory proteins mAPM-1, LRF, KLHL10 that belong to a subgroup of human POZ-domain proteins, and showed that the POZ-domains of mAPM-1, LRF and KLHL10 could form both homodimers and heterodimers. This new method is a highly efficient, sensitive and specific assay method for protein-protein interaction in vivo.