title: A Comparison of Two-Hybrid Approaches for Detecting Protein-Protein Interactions.
abstract: Two-hybrid systems are one of the most popular, preferred, cost effective, and scalable in vivo genetic approaches for screening protein-protein interactions. A number of variants of yeast and bacterial two-hybrid systems exist, rendering them ideal for modern, flexible proteomics-driven studies. For mapping protein interactions at genome scales (that is, constructing an interactome), the yeast two-hybrid system has been extensively tested and is preferred over bacterial two-hybrid systems, given that users have created more resources such as a variety of vectors and other modifications. Each system has its own advantages and limitations and thus needs to be compared directly. For instance, the bacterial two-hybrid method seems a better fit than the yeast two-hybrid system to screen membrane-associated proteins. In this chapter, we provide detailed protocols for yeast and bacterial two-hybrid systems as well as a comparison of outcomes for each approach using our own and published data.
authors.affiliation: Center for the Study of Biological Complexity, Virginia Commonwealth University, Richmond, VA, United States. Electronic address: jitender@vcu.edu.
authors.forename: J
authors.initials: J
authors.lastname: Mehla
authors.affiliation: Center for the Study of Biological Complexity, Virginia Commonwealth University, Richmond, VA, United States.
authors.forename: J H
authors.initials: JH
authors.lastname: Caufield
authors.affiliation: Center for the Study of Biological Complexity, Virginia Commonwealth University, Richmond, VA, United States.
authors.forename: N
authors.initials: N
authors.lastname: Sakhawalkar
authors.affiliation: Center for the Study of Biological Complexity, Virginia Commonwealth University, Richmond, VA, United States. Electronic address: peter@uetz.us.
authors.forename: P
authors.initials: P
authors.lastname: Uetz
mesh_terms: D004926:Escherichia coli; D011485:Protein Binding; D060066:Protein Interaction Maps; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae; D014169:Transformation, Bacterial; D020798:Two-Hybrid System Techniques
chemical_list: D011993:Recombinant Fusion Proteins
keywords: Bacterial two-hybrid protocol; In vivo genetic methods; Interactome; Protein–protein interactions; Yeast two-hybrid protocol