A two-hybrid assay fluorescent (F2H) for direct visualization of protein interactions in living cells

摘要

An in vitro method for detecting protein-protein interactions comprising: (a) expressing in a eukaryotic cell a first fusion protein comprising (i) a (poly) peptide that, when expressed in a cell, accumulates at sites different in the cell nucleus; and (ii) a (poly) peptide that specifically binds the green fluorescent protein (GFP) (b) expressing in the same cell a second fusion protein comprising (i) GFP; and (ii) a (poly) peptide lure (c) expressing in the same cell a third fusion protein comprising (i) a (poly) fluorescent peptide, whose wavelength excitation and / or emission differs from that of GFP ; and (ii) a (poly) peptide prey (d) detecting the fluorescence emission of the fluorescent parts of the second and the third fusion protein in the cell upon excitation, wherein a co-localization of the fluorescence emission of both fusion proteins in the cell nucleus is indicative of an interaction of said (poly) peptide bait and prey.