Yeast two-hybrid system for studying protein-protein interactions--stage 2: Transforming and characterizing the library.
An important element in the characterization of the function of a protein is the identification of other proteins with which it interacts. A powerful genetic strategy for this purpose, termed the "yeast two-hybrid system," uses transcriptional reporters in yeast to indirectly reflect the interaction between two proteins. The term "two-hybrid" derives from the two classes of chimeric, or "hybrid," proteins used in each screen. The first, commonly referred to as the "bait," is a fusion of a protein of interest "x" with a DNA-binding domain (DBD-x). The second, sometimes called the "prey," is a fusion of a cDNA library "y" to a transcriptional activation domain (AD-y). Together, DBD-x and AD-y provide the basis of the detection system. The two-hybrid approach has gained wide popularity because of the relative ease and speed with which it can be used to identify novel protein-protein interactions and to analyze known interactions. The second stage of the method, described in this protocol, includes the transformation of yeast with a cDNA library, followed by library characterization. It can be performed in parallel with construction of a bait protein (stage 1).
Serebriiskii
Ilya
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D015723:Gene Library; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D010957:Plasmids; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic; D020798:Two-Hybrid System Techniques
D011506:Proteins