Human Molecular Genetics, Vol 7, 1029-1032, Copyright © 1998 by Oxford University Press
H Schwartz, CP Alvares, MB White and S Fields
Yeast-based assays have been developed to detect inactivating mutations in
human genes, but these assays generally rely on the human protein having a
biological function in yeast. We describe a simple method to detect
mutations by virtue of their ability to abolish a protein- protein
interaction in the yeast two-hybrid assay. By the use of direct
recombinational cloning in yeast of a reverse transcription-PCR product
followed by a simple growth selection this method distinguished both
homozygous and heterozygous mutations in the p53 tumor suppressor gene.
This approach should be applicable to many human genes whose encoded
proteins have suitable partners in the two-hybrid assay.
ARTICLES
Mutation detection by a two-hybrid assay
Department of Genetics, University of Washington, Box 357360, Seattle, WA 98195, USA.
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