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Two-Hybrid Systems Methods and Protocols
Edited by  Paul N. MacDonald


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Preface........................................................................................... v
Contributors .................................................................................. xi
PART I GENERAL METHODS
1 The Two-Hybrid System: A Personal View
Stanley Fields and Paul L. Bartel .............................................. 3
2 Growth and Maintenance of Yeast
Lawrence W. Bergman ................................................................ 9
3 Media Formulations for Various Two-Hybrid Systems
Michael Saghbini, Denise Hoekstra, and Jim Gautsch ........ 15
4 Yeast Two-Hybrid Vectors and Strains
Philip James ............................................................................... 41
5 High-Efficiency Transformation of Plasmid DNA into Yeast
Robin A. Woods and R. Daniel Gietz ...................................... 85
6 Qualitative and Quantitative Assessment of Interactions
Monica M. Montano ................................................................... 99
7 Strategies for Rescuing Plasmid DNA
from Yeast Two-Hybrid Colonies
Alyson Byrd and Ren¨¦ St-Arnaud ......................................... 107
PART II FALSE POSITIVES
8 Two-Hybrid System and False Positives:
Approaches to Detection and Elimination
Ilya G. Serebriiskii and Erica A. Golemis ............................. 123
9 Confirming Yeast Two-Hybrid Protein Interactions
Using In Vitro Glutathione-S-Transferase Pulldowns
Dennis M. Kraichely and Paul N. MacDonald ...................... 135
10 Two-Hybrid Interactions Confirmed
by Coimmunoprecipitation of Epitope-Tagged Clones
Louie Naumovski ..................................................................... 151
Contents
PART III APPLICATIONS
11 Smad Interactors in Bone Morphogenetic Protein Signaling
Xiangli Yang and Xu Cao ........................................................ 163
12 Protein Interactions Important in Eukaryotic
Translation Initiation
Katsura Asano and Alan G. Hinnebusch.............................. 179
13 Steroid Receptor and Ligand-Dependent Interaction
with Coactivator Proteins
Sergio A. Oñate........................................................................ 199
14 Interaction of Cellular Apoptosis Regulating Proteins
with Adenovirus Anti-apoptosis Protein E1B-19K
Thirugnana Subramanian and G. Chinnadurai.................... 211
PART IV ALTERNATIVE STRATEGIES
15 Mammalian Two-Hybrid Assays: Analyzing Protein-Protein
Interactions in the Transforming Growth Factor-¦Â
Signaling Pathway
Xin-Hua Feng and Rik Derynck.............................................. 221
16 One-Hybrid Systems for Detecting Protein-DNA Interactions
Mary Kate Alexander, Brenda D. Bourns,
and Virginia A. Zakian ......................................................... 241
17 The Split-Hybrid System: Uncoding Multiprotein Networks
and Defining Mutations That Affect Protein Interactions
Phyllis S. Goldman, Anthony J. DeMaggio,
Richard H. Goodman, and Merl F. Hoekstra .................... 261
18 Three-Hybrid Screens: Inducible Third-Party Systems
Björn Sandrock, Franck Tirode, and Jean-Marc Egly......... 271
19 Three-Hybrid Screens for RNA-Binding Proteins:
Proteins Binding 3' End of Histone mRNA
Zbigniew Dominski and William F. Marzluff......................... 291
20 Membrane Recruitment Systems for Analysis
of Protein¨CProtein Interactions
Ami Aronheim .......................................................................... 319
Index ..........................................................................................329


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