MacDonald | Two-Hybrid Systems | Buch | 978-1-61737-210-0 | sack.de

Buch, Englisch, Band 177, 336 Seiten, Previously published in hardcover, Format (B × H): 152 mm x 229 mm, Gewicht: 516 g

Reihe: Methods in Molecular Biology

MacDonald

Two-Hybrid Systems

Methods and Protocols
1. Auflage. Softcover version of original hardcover Auflage 2001
ISBN: 978-1-61737-210-0
Verlag: Humana Press

Methods and Protocols

Buch, Englisch, Band 177, 336 Seiten, Previously published in hardcover, Format (B × H): 152 mm x 229 mm, Gewicht: 516 g

Reihe: Methods in Molecular Biology

ISBN: 978-1-61737-210-0
Verlag: Humana Press


Many, if not all, essential biological processes require selective interactions between proteins. Complex signaling systems require sequential, ordered protein–protein interactions at essentially all levels of the signaling cascade. For example, peptide hormones interact with selective membrane receptor proteins, and autophosphorylation of the receptor then recruits other key regulatory proteins that initiate kinase cascades in which each phosphorylation event requires selective recognition of the protein substrate. The ultimate signaling effect, in many cases, is the regulation of RNA polymerase II-directed transcr- tion in the nucleus, a process that involves numerous, multiprotein complexes important for transcription initiation, elongation, termination, and reinitiation. Defining, characterizing, and understanding the relevance of these protein– protein interactions is an arduous task, but substantial inroads have been made over the past 20 years. The development of more recent methodologies, such as mammalian expression systems, immunopurification schemes, expression cloning strategies, surface plasmon resonance (BiaCore), and nanosequencing technologies, has contributed a wealth of new insights into these complex multiprotein mechanisms and clearly accelerated the discovery process. Arguably, the yeast two-hybrid system has been one of the predominant and most powerful tools in this discovery process.

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Part I. General Methods

The Two-Hybrid System: A Personal View
Stanley Fields and Paul L. Bartel

Growth and Maintenance of Yeast
Lawrence W. Bergman

Media Formulations for Various Two-Hybrid Systems
Michael Saghbini, Denise Hoekstra, and Jim Gautsch

Yeast Two-Hybrid Vectors and Strains
Philip James

High-Efficiency Transformation of Plasmid DNA into Yeast
Robin A. Woods and R. Daniel Gietz

Qualitative and Quantitative Assessment of Interactions
Monica M. Montano

Strategies for Rescuing Plasmid DNA from Yeast Two-Hybrid Colonies,
Alyson Byrd and René St-Arnaud

Part II. False Positives

Two-Hybrid System and False Positives: Approaches to Detection and Elimination
Ilya G. Serebriiskii and Erica A. Golemis

Confirming Yeast Two-Hybrid Protein Interactions Using In Vitro Glutathione-S-Transferase Pulldowns
Dennis M. Kraichely and Paul N. MacDonald

Two-Hybrid Interactions Confirmed by Coimmunoprecipitation of Epitope-Tagged Clones
Louie Naumovski

Part III. Applications

Smad Interactors in Bone Morphogenetic Protein Signaling
Xiangli Yang and Xu Cao

Protein Interactions Important in Eukaryotic Translation Initiation
Katsura Asano and Alan G. Hinnebusch

Steroid Receptor and Ligand-Dependent Interaction with Coactivator Proteins
Sergio A. Oñate

Interaction of Cellular Apoptosis Regulating Proteins with Adenovirus Anti-Apoptosis Protein E1B-19K
Thirugnana Subramanian and G. Chinnadurai

Part IV. Alternative Strategies

Mammalian Two-Hybrid Assays: Analyzing Protein-Protein Interactions in the Transforming Growth Factor-_ Signaling Pathway
Xin-Hua Feng and Rik Derynck

One-Hybrid Systems for Detecting Protein-DNA Interactions
Mary Kate Alexander, Brenda D. Bourns, and Virginia A.Zakian

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

Three-Hybrid Screens: Inducible Third-Party Systems
Björn Sandrock, Franck Tirode, and Jean-Marc Egly

Three-Hybrid Screens for RNA-Binding Proteins: Proteins Binding 3' End of Histone mRNA
Zbigniew Dominski and William F. Marzluff

Membrane Recruitment Systems for Analysis of Protein-Protein Interactions
Ami Aronheim



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