Elsevier

Methods in Enzymology

Volume 254, 1995, Pages 241-263
Methods in Enzymology

[16] Analyzing protein-protein interactions using two-hybrid system

https://doi.org/10.1016/0076-6879(95)54018-0Get rights and content

Publisher Summary

This chapter presents the analysis of protein–protein interactions using two-hybrid system. Protein-protein interactions play a critical role in most biological processes. The studies defining domains of proteins that are responsible for specific interactions have contributed significantly to unraveling the mechanisms of tumorigenesis. The two-hybrid system is a yeast-based genetic assay for detecting protein– protein interactions in vivo. It can be used to establish interactions between two known proteins or to search genomic or cDNA libraries for proteins that interact with a target protein. For this latter application, the gene encoding the protein that interacts with a target protein is immediately available on a plasmid, which is not the case for many biochemical methods to detect interacting proteins. The two-hybrid system has also been used to define the protein domains that mediate an interaction and to identify specific residues that are involved in a protein–protein interaction. The chapter also discusses the basis for this method and presents the protocols that are necessary to use this system.

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