NDLI: Yeast two-hybrid system for studying protein-protein interactions--stage 4: Isolation of library plasmid insert and second confirmation of positive interactions.

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Yeast two-hybrid system for studying protein-protein interactions--stage 1: Construction and characterization of a bait protein.

Yeast two-hybrid system for studying protein-protein interactions--stage 2: Transforming and characterizing the library.

Yeast two-hybrid system for studying protein-protein interactions--stage 3: Screen for interacting proteins.

Yeast two-hybrid system for studying protein-protein interactions--stage 4: Isolation of library plasmid insert and second confirmation of positive interactions.

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Yeast two-hybrid system for studying protein-protein interactions--stage 4: Isolation of library plasmid insert and second confirmation of positive interactions.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Serebriiskii, Ilya
Abstract	An important element in the characterization of the function of a protein is the identification of other proteins with which it interacts. A powerful genetic strategy for this purpose, termed the 'yeast two-hybrid system,' uses transcriptional reporters in yeast to indirectly reflect the interaction between two proteins. The term 'two-hybrid' derives from the two classes of chimeric, or 'hybrid,' proteins used in each screen. The first, commonly referred to as the 'bait,' is a fusion of a protein of interest 'x' with a DNA-binding domain (DBD-x). The second, sometimes called the 'prey,' is a fusion of a cDNA library 'y' to a transcriptional activation domain (AD-y). Together, DBD-x and AD-y provide the basis of the detection system. The two-hybrid approach has gained wide popularity because of the relative ease and speed with which it can be used to identify novel protein-protein interactions and to analyze known interactions. Stage 4, described here, outlines a series of first-order and subsequent control experiments designed to establish whether an interacting protein is likely to be biologically significant. The number of positive interactions obtained will vary drastically from bait to bait. Subsequent processing methods will depend on the number initially obtained and on the preference of the individual investigator.
File Format	HTM / HTML
ISSN	19403402
Issue Number	5
Volume Number	2010
e-ISSN	15596095
Journal	Cold Spring Harbor Protocols
Language	English
Publisher	Cold Spring Harbor Laboratory Press
Publisher Date	2010-05-01
Publisher Place	United States
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Discipline Clinical Laboratory Techniques Protein Interaction Mapping Methods Two-hybrid System Techniques Cloning, Molecular Dna, Fungal Genetics Isolation & Purification Gene Library Gene Transfer Techniques Genetic Vectors Plasmids Protein Binding Proteins Metabolism Recombination, Genetic Transformation, Genetic Journal Article
Content Type	Text
Resource Type	Article
Subject	Biochemistry, Genetics and Molecular Biology

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