Selective detection of inactivating mutations of the p53 tumour suppressor gene: Development of a new functional assay in yeast
- CHU de Rouen (France)
- ISREC, Epalinges (Switzerland)
Alterations of the p53 tumour suppressor gene in human cancer are mainly missense mutations. We have previously described a functional assay in Saccharomyces cerevisiae based on a HIS3 reporter system, which allows the selective detection of mutations which have inactivated the transcriptional activity of p53. We have now developed a simpler reporter system in which yeast change color according to their p53 status. We used an ade2-1 strain which contains an intergrated copy of the ADE2 open reading frame controlled by a p53-sensitive promoter. In the absence of wild-type p53, the strain is red due to the accumulation of an intermediate in adenine metabolism. As in the previous assay, we transform PCR-amplified p53 cDNA directly into the strain and clone them into a p53 expression vector by homologous recombination in vivo. Colonies that contain wild-type p53 overcome the block in adenine synthesis and become white. After two days, the white/red system allows to distinguish very easily wild-type and mutant p53 alleles. This new assay is a simple and rapid method to detect inactivating p53 mutations in clinical samples.
- OSTI ID:
- 133397
- Report Number(s):
- CONF-941009-; ISSN 0002-9297; TRN: 95:005313-0125
- Journal Information:
- American Journal of Human Genetics, Vol. 55, Issue Suppl.3; Conference: 44. annual meeting of the American Society of Human Genetics, Montreal (Canada), 18-22 Oct 1994; Other Information: PBD: Sep 1994
- Country of Publication:
- United States
- Language:
- English
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