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

DOIResolve DOI: http://doi.org/10.1385/1-59259-356-9:269
AuthorSearch for:
EditorSearch for: M. Terrian, David
TypeBook Chapter
Book titleCancer Cell Signaling: Methods and Protocols
Series titleMethods in Molecular Biology; Volume 218
ISBN978-1-58829-075-5
Pages269276; # of pages: 8
Subjectpha; cancer research; protein; protein-protein interactions; mutagenesis; screening; two-hybrid; interaction trap
AbstractAcquiring 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
PublisherHumana Press Inc.
LanguageEnglish
AffiliationNational Research Council Canada; NRC Biotechnology Research Institute
Peer reviewedNo
NRC number44804
NPARC number3539165
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Record identifierbc13845f-8e84-4666-8bdc-a42d7fede545
Record created2009-03-01
Record modified2016-05-09
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