NDLI: Coupled yeast 2-hybrid-mammalian 2-hybrid reading-frame-independent and site-specific recombinational cloning vector system.

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Coupled yeast 2-hybrid-mammalian 2-hybrid reading-frame-independent and site-specific recombinational cloning vector system.

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Coupled yeast 2-hybrid-mammalian 2-hybrid reading-frame-independent and site-specific recombinational cloning vector system.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Maier, Christina J. Maier, Richard H. Hintner, Helmut Bauer, Johann W. Onder, Kamil
Description	Country affiliation: Austria Author Affiliation: Maier CJ ( Department of Cell Biology, University of Salzburg/Naturwissenschaftliche Fakultät, Salzburg, Austria. maier.christina@rocketmail.com)
Abstract	The yeast 2-hybrid (Y2H) system is a powerful method for identifying protein-protein interactions (PPIs), requiring minimal prior information of the putative interactors. Currently available automated versions of the Y2H system are sufficiently developed to allow facile genome-wide PPI screening to compile extensive inter-actome data. A limitation of the Y2H approach, however, is that all primary hits have to be technically verified and biologically evaluated by complementary methods, which is time-consuming, costly, and laborious. Furthermore, the yeast intracellular environment can lead to spurious results for proteins of other organisms, for example because of differences in post-translational modifications or the presence/absence of bridging proteins. Many researchers now confirm PPIs found in the Y2H system by retesting the candidates in the mammalian 2-hybrid system (M2H). However, although such combined Y2H-M2H testing is desirable and perhaps necessary, recloning of Y2H candidates into M2H vectors, especially on a large scale, is time-consuming and costly. To address this shortcoming, we introduce here a new site-specific recombination-capable M2H vector system that is fully compatible with the site-specific Y2H system that we recently described in Biotechniques [2008;45(3):235-244]. The results show that the new vectors are: (a) Gateway®, compatible and suitable for fast cloning; (b) fully functional in the M2H system without influencing the capacity of the selection system or creating autoactivators; and (c) directly compatible with the existing site-specific Y2H vector system, as demonstrated by confirmation of Y2H PPI candidates in the M2H system.
File Format	HTM / HTML
ISSN	1540658X
Issue Number	5
Volume Number	8
e-ISSN	15578127
Journal	ASSAY and Drug Development Technologies
Language	English
Publisher	Mary Ann Liebert, Inc.
Publisher Date	2010-10-01
Publisher Place	United States
Access Restriction	Subscribed
Subject Keyword	Discipline Analytical Discipline Pharmacology Discipline Chemistry Techniques Cloning, Molecular Genetic Vectors Two-hybrid System Techniques Animals Antigens, Polyomavirus Transforming Metabolism Cell Line Gene Library Genetic Techniques Hek293 Cells Humans Protein Binding Protein Interaction Domains And Motifs Reading Frames Yeasts Genetics Journal Article
Content Type	Text
Resource Type	Article
Subject	Drug Discovery Molecular Medicine

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