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Screening patients for heterozygous p53 mutations using a functional assay in yeast

Nature Genetics volume 5, pages 124–129 (1993)Cite this article

Abstract

Inherited mutations of the p53 gene significantly increase the risk of developing diverse malignancies, and germline p53 mutations can be detected by assaying the transcriptional activity of the p53 protein in mammalian cells. Here we describe a method starting with lymphocytes that allows detection of germline p53 mutations by ‘functional’ analysis of p53 protein expressed in Saccharomyces cerevisiae. The p53 PCR products are directly cloned into yeast expression vectors in vivo and subsequently tested for transcriptional activity in a simple growth assay. This technique, functional analysis of separated alleles in yeast (FASAY), requires only a few steps, can be automated readily and should permit screening for germline or somatic heterozygous mutations in any gene whose function can be monitored in yeast.

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Author notes

  1. Thierry Frebourg

    Present address: Unité de Génétique Moléculaire, CHU de Rouen, 76000, Rouen, France

Authors and Affiliations

  1. Massachusetts General Hospital Cancer Center, Charlestown, Massachusetts, 02129, USA

    Chikashi Ishioka, Thierry Frebourg, Yu-Xin Yan, Marc Vidal & Stephen H. Friend

  2. Department of Clinical Oncology, Institute of Development, Aging and Cancer (IDAC), Tohoku University, Sendai, 980, Japan

    Chikashi Ishioka

  3. Department of Pediatrics, Division of Hematology-Oncology, The Children's Hospital, Dana-Farber Institute, Harvard Medical School, Boston, Massachusetts, 02114, USA

    Stephen H. Friend

  4. Swiss Institute for Experimental Cancer Research (ISREC), 1066, Epalinges, Switzerland

    Susanne Schmidt & Richard Iggo

Authors
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Ishioka, C., Frebourg, T., Yan, YX. et al. Screening patients for heterozygous p53 mutations using a functional assay in yeast. Nat Genet 5, 124–129 (1993). https://doi.org/10.1038/ng1093-124

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  • DOI: https://doi.org/10.1038/ng1093-124

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