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  A stringent yeast two-hybrid matrix screening approach for protein-protein interaction discovery

Worseck, J., Grossmann, A., Weimann, M., Hegele, A. K., & Stelzl, U. (2012). A stringent yeast two-hybrid matrix screening approach for protein-protein interaction discovery. Methods in Molecular Biology, 812, 63-87. doi:10.1007/978-1-61779-455-1_4.

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Item Permalink: https://hdl.handle.net/11858/00-001M-0000-000E-E7B4-4 Version Permalink: https://hdl.handle.net/11858/00-001M-0000-000E-E7B5-2
Genre: Journal Article

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 Creators:
Worseck, Josephine1, Author           
Grossmann, Arndt2, Author           
Weimann, Mareike1, Author           
Hegele, Anna K.1, Author           
Stelzl, Ulrich2, Author           
Affiliations:
1Independent Junior Research Groups (OWL), Max Planck Institute for Molecular Genetics, Max Planck Society, Berlin, Germany, ou_1433554              
2Molecular Interaction Networks (Ulrich Stelzl), Independent Junior Research Groups (OWL), Max Planck Institute for Molecular Genetics, Max Planck Society, Berlin, Germany, ou_1479660              

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Free keywords: Escherichia coli/genetics Proteins/*analysis/*metabolism Transformation, Bacterial *Two-Hybrid System Techniques Yeasts/genetics/growth & development beta-Galactosidase/metabolism
 Abstract: The yeast two-hybrid (Y2H) system is currently one of the most important techniques for protein-protein interaction (PPI) discovery. Here, we describe a stringent three-step Y2H matrix interaction approach that is suitable for systematic PPI screening on a proteome scale. We start with the identification and elimination of autoactivating strains that would lead to false-positive signals and prevent the identification of interactions. Nonautoactivating strains are used for the primary PPI screen that is carried out in quadruplicate with arrayed preys. Interacting pairs of baits and preys are identified in a pairwise retest step. Only PPI pairs that pass the retest step are regarded as potentially biologically relevant interactions and are considered for further analysis.

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Language(s): eng - English
 Dates: 2012
 Publication Status: Issued
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1007/978-1-61779-455-1_4
 Degree: -

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Title: Methods in Molecular Biology
  Other : Methods Mol. Biol.
Source Genre: Journal
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Publ. Info: Clifton, N.J. : Humana Press
Pages: - Volume / Issue: 812 Sequence Number: - Start / End Page: 63 - 87 Identifier: ISSN: 1064-3745
CoNE: https://pure.mpg.de/cone/journals/resource/954927725544