A Comparison of Two-Hybrid Approaches for Detecting Protein–Protein Interactions [chapter]

J. Mehla, J.H. Caufield, N. Sakhawalkar, P. Uetz
2017 Methods in Enzymology  
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,
more » ... n 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.
doi:10.1016/bs.mie.2016.10.020 pmid:28137570 pmcid:PMC5737774 fatcat:vrsskowwrva47lef4xlqtld7si