FLUORESCENT TWO-HYBRID (F2H) ASSAY FOR DIRECT VISUALIZATION OF PROTEIN INTERACTIONS IN LIVING CELLS
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Nov 30, 2016
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Jan 9, 2008
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Abstract
The present invention relates to a method for detecting protein-protein interactions by assessing interaction in a eukaryotic cell of a first fusion protein that specifically binds to GFP and accumulates at distinct sites in the nucleus of the cell or interacts with structures accumulated at distinct sites in the nucleus of the cell; a second fusion protein comprising GFP and a bait (poly)peptide; and a third fusion protein comprising a fluorescent (poly)peptide having an excitation and/or emission wavelength that differs from that of GFP and a prey (poly)peptide. The emissions from the fluorescent parts of the fusion proteins are observed. Co-localization of the emissions from both fluorescent fusion proteins indicates interaction of the bait and the prey (poly)peptide. Methods for identifying a compound modulating the interaction of two (poly)peptides and methods of determining the relative strength of the interaction of two proteins with a third protein.
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