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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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 Urheber:
Worseck, Josephine1, Autor           
Grossmann, Arndt2, Autor           
Weimann, Mareike1, Autor           
Hegele, Anna K.1, Autor           
Stelzl, Ulrich2, Autor           
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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Schlagwörter: Escherichia coli/genetics Proteins/*analysis/*metabolism Transformation, Bacterial *Two-Hybrid System Techniques Yeasts/genetics/growth & development beta-Galactosidase/metabolism
 Zusammenfassung: 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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Sprache(n): eng - English
 Datum: 2012
 Publikationsstatus: Erschienen
 Seiten: -
 Ort, Verlag, Ausgabe: -
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 Art der Begutachtung: Expertenbegutachtung
 Identifikatoren: DOI: 10.1007/978-1-61779-455-1_4
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Titel: Methods in Molecular Biology
  Andere : Methods Mol. Biol.
Genre der Quelle: Zeitschrift
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Ort, Verlag, Ausgabe: Clifton, N.J. : Humana Press
Seiten: - Band / Heft: 812 Artikelnummer: - Start- / Endseite: 63 - 87 Identifikator: ISSN: 1064-3745
CoNE: https://pure.mpg.de/cone/journals/resource/954927725544