Funkční analýza separovaných alel v kvasinkách: spolehlivá metoda k určení statutu nádorového supresoru p53
| Title in English | Functional analysis of separated alleles in yeast: a reliable method for assesment of the p53 status |
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| Authors | |
| Year of publication | 2005 |
| Type | Article in Proceedings |
| Conference | Dny diagnostické, prediktivní a experimentální onkologie |
| MU Faculty or unit | |
| Citation | |
| Field | Oncology and hematology |
| Keywords | FASAY; p53; p53 transactivation; gap repair |
| Description | FASAY is a sensitive semi-quantitative method of detection of p53 mutations in clinical material, such as tumor tissues, peripheral blood cells, etc. The p53 is derived from mRNA of analyzed material by RT-PCR and its transactivation ability is tested in yeast cells by using of ADE2 reporter gene. Transactivation by p53 is assessed simply according to the color of yeast colonies growing on agar plates with lovele of adenine. Yeast cells form either large white colonies if the reporter was activated by functional p53 or small red colonies if the reporter was not activated. FASAY possesses several advantages: (1) it analyzes both alleles of the p53 gene separately, (2) its high sensitivity allows detection of about 10% of p53 mutation harboring cells among mostely negative cells, (3) it can distinguish fully inactivating p53 mutations from partially inactivating ones, (4) it analyzes substantial part of the p53 gene, (5) it allows preparation of template useful for sequencing of the p53 gene. |
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