Using the yeast two-hybrid system to identify interacting proteins.
The yeast two-hybrid system is a powerful technique for studying protein-protein interactions. Two proteins are separately fused to the independent DNA-binding and transcriptional activation domains of the Gal4p transcription factor. If the proteins interact, they reconstitute a functional Gal4p that activates expression of reporter gene(s). In this way, two individual proteins may be tested for their ability to interact, and a transcriptional readout can be measured to detect this interaction. Furthermore, novel interacting partners can be found by screening a single protein or domain against a library of other proteins using this system. It is this latter feature-the ability to search for interacting proteins without any prior knowledge of the identity of such proteins-that is the most powerful application of the two-hybrid technique.
Miller
John
J
Department of Genome Sciences, University of Washington, Seattle, WA, USA.
Stagljar
Igor
I
D006801:Humans; D025941:Protein Interaction Mapping / Q000379:methods*; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000235:genetics*
D011506:Proteins; D014157:Transcription Factors