Development of yeast color assay for screening of genetic mutation

1. APC酵母显色试验：我们开发了一种灵敏的基于酵母的终止密码子试验，用于筛选APC基因的几乎整个编码区。通过这种方法，我们已经能够在57%的结直肠癌、18%的乳腺癌中检测到APC基因的体细胞突变，而在非小细胞肺癌中没有检测到。突变APC产生红酵母菌落. p53显性负性测定：我们使用基于酵母的测定法检测了各种p53突变体的显性负性潜力，其中野生型和突变型p53均有效表达。我们总共检测了106个p53突变体。显性阴性p53产生粉红色酵母菌集落. PTEN终止密码子测定：我们已经开发了一种基于酵母的检测PTEN无义突变的方法（显示为红酵母菌落），并将其应用于总共42个星形细胞肿瘤。有趣的是，在18个胶质母细胞瘤中的10个中发现了粉红色菌落。粉红色菌落的序列分析揭示了psi PTEN cDNA的序列。

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 nonsmall 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.