Chapter 14. Analysis of RNA-protein interactions using a yeast three-hybrid system.
RNA-protein interactions play an essential role in the maturation and regulation of RNAs within eukaryotic organisms. The three-hybrid system provides a simple, yet powerful means to study RNA-protein interactions within the eukaryote Saccharomyces cerevisiae. This chapter describes the basis of the system and applications in both examining specific RNA-protein interactions and screening libraries for novel interactions. We provide a detailed discussion on affinity versus reporter output, variations on library screening (e.g., randomization studies), some adaptations of the system, and updated reagents and protocols.
Stumpf
Craig R
CR
Program in Cellular and Molecular Biology, University of Wisconsin-Madison, Madison, Wisconsin, USA.
Opperman
Laura
L
Wickens
Marvin
M
D011485:Protein Binding / Q000235:genetics; D012313:RNA / Q000235:genetics / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
D016601:RNA-Binding Proteins; D012313:RNA