Production of ligand-specific mutants using a yeast two-hybrid mating assay

DOI	Resolve DOI: https://doi.org/10.1385/1-59259-356-9:269
Author	Search for: Nantel, André¹
Editor	Search for: M. Terrian, David
Name affiliation	National Research Council of Canada. NRC Biotechnology Research Institute
Format	Text, Book chapter
Book title	Cancer Cell Signaling: Methods and Protocols
Series title	Methods in Molecular Biology; Volume 218
ISBN	978-1-58829-075-5
Pages	269–276
Subject	pharmaceutical; cancer research; protein; protein-protein interactions; mutagenesis; screening; two-hybrid; interaction trap
Abstract	Acquiring functional knowledge from protein-protein interaction studies often necessitates the production of binding-impaired mutants and the study of their effects in biological systems. In many cases, multiple ligands compete for binding to the same protein domain and it becomes useful to produce specific mutations that prevent binding to one ligand but not to the others. We have combined PCR mutagenesis and a two hybrid mating assay to produce a screening strategy that has already proven useful in isolating ligand-specific mutants of the Grb10 and Raf-1 signaling proteins.
Publication date	2003
Publisher	Humana Press Inc. Humana Press Inc.
Language	English
NRC number	NRC-BRI-44804
NPARC number	3539165
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Record identifier	bc13845f-8e84-4666-8bdc-a42d7fede545
Record created	2009-03-01
Record modified	2020-06-12