J-GLOBAL ID:200904055234900119  Research Project code:0250005283 Update date:Dec. 20, 2002

Development of yeast-based method for detecting gene mutations

Study period:1998 - 2003
Research overview:
1. APC yeast color assay: We have developed a sensitive yeast-based stop codon assay for screening almost the entire coding region of the APC gene. By this method we have been able to detect somatic mutations of the APC gene in 57% of colorectal cancers, 18% of breast cancers, and none in non-small cell lung cancers. Mutant APC generated red yeast colonies. 2. p53 dominant-negative assay: We examined the dominant-negative potential of various p53 mutants using a yeast-based assay in which both wild-type and mutant p53 were efficiently expressed. We have tested a total of 106 p53 mutants. Dominant-negative p53 generated pink yeast colonies. 3. PTEN stop codon assay: We have developed a yeast-based assay for the detection of PTEN nonsense mutation (shown as red yeast colonies), and applied it to a total of 42 astrocytic tumors. Interestingly pink colonies were found in 10 of 18 glioblastomas. Sequence analysis of the pink colonies revealed a sequence of psi PTEN cDNA.
Keywords (6):
yeast assay ,  tumor suppressor gene ,  oncogene ,  p53 ,  APC ,  PTEN
Research program: Grant-in-Aid for Scientific Research
Ministry with control over the research :
Ministry of Education, Culture, Sports, Science and Technology
Research budget: 2001: \15,000,000

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