Mapping protein-protein interactions using yeast two-hybrid assays.
Yeast two-hybrid (Y2H) screens are an efficient system for mapping protein-protein interactions and whole interactomes. The screens can be performed using random libraries or collections of defined open reading frames (ORFs) called ORFeomes. This protocol describes both library and array-based Y2H screening, with an emphasis on array-based assays. Array-based Y2H is commonly used to test a number of "prey" proteins for interactions with a single "bait" (target) protein or pool of proteins. The advantage of this approach is the direct identification of interacting protein pairs without further downstream experiments: The identity of the preys is known and does not require further confirmation. In contrast, constructing and screening a random prey library requires identification of individual prey clones and systematic retesting. Retesting is typically performed in an array format.
Center for the Study of Biological Complexity, Virginia Commonwealth University, Richmond, Virginia 23284.
Jitender
J
Mehla
Center for the Study of Biological Complexity, Virginia Commonwealth University, Richmond, Virginia 23284.
J Harry
JH
Caufield
Center for the Study of Biological Complexity, Virginia Commonwealth University, Richmond, Virginia 23284.
Peter
P
Uetz
D005820:Genetic Testing / Q000379:methods*; D060066:Protein Interaction Maps; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D020798:Two-Hybrid System Techniques
D011994:Recombinant Proteins