Journal Article

Mutation Detection by a Two-Hybrid Assay

Hillel Schwartz, Christopher P. Alvares, Marga B. White and Stanley Fields

in Human Molecular Genetics

Volume 7, issue 6, pages 1029-1032
Published in print June 1998 | ISSN: 0964-6906
Published online June 1998 | e-ISSN: 1460-2083 | DOI: http://dx.doi.org/10.1093/hmg/7.6.1029
Mutation Detection by a Two-Hybrid Assay

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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 homo-zygous 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.

Journal Article.  2831 words.  Illustrated.

Subjects: Genetics and Genomics

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