The Yeast Two-hybrid System

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Paul L. Bartel, Stanley Fields
Oxford University Press, 1997 - Science - 344 pages
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Biologists have already identified thousands of new gene sequences, and genome sequencing efforts are speeding up the discovery process even further. With this explosion of sequence information comes the need to understand how genes work in concert in order to fulfill the cells functions. The yeast two-hybrid system--used to identify protein-protein interactions--is one of the most powerful and versatile methods for characterizing a protein's function. It has become an essential tool for both academic researchers and those in biotechnology and pharmaceutical companies. This volume presents work by pioneers in the field and is the first publication devoted solely to the yeast two-hybrid system. It includes detailed protocols, practical advice on troubleshooting, and suggestions for future development. In addition, it explains how to construct an activation domain hybrid library, how to identify mutations that disrupt an interaction, and how to use the system in mammalian cells. Many of the contributors have developed new applications and variations of the technique. Chapter topics include characterizing hormone/receptor complexes, identifying peptide ligands, analyzing interactions mediated by protein modifications, and dissecting the cell cycle and other complex genetic networks. The Yeast Two-Hybrid System is the single complete resource for scientists interested in this powerful research method.
 

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Contents

A Personal View
3
Analyzing Interactions with the TwoHyhrid System
6
The TwoHyhrid Screen
11
Searching for Interacting Proteins with
43
Constructing an Activation DomainFusion Lihrary
73
The Reverse TwoHyhrid System
109
Dissecting the Biological Relevance of Multiple Interactions
149
Investigation of LigandReceptor Interactions and
158
l2 TwoHyhrid Analysis of Genetic Regulatory Networks
197
Part HI Variations on the TwoHybrid Theme
217
l4 Detection of ProteinProtein Interactions Dependent
233
l5 Protein Interactions Mediated hy Small Molecule Ligands
259
l6 Use of a Comhinatorial Peptide Lihrary in
273
Related Methodologies
287
l8 A ThreeHyhrid System to Detect and Analyze RNAProtein
298
a Sensor of Protein Interactions In Vivo
316

l0 TwoHyhrid Analysis of ProteinProtein Interactions in
173
TwoHyhrid Screening and the Cell Cycle
183

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About the author (1997)

Paul L. Bartel is at Myriad Genetics, Salt Lake City. Stanley Fields is at University of Washington.

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