Development of yeast color assay for screening of genetic mutation

开发用于筛选基因突变的酵母颜色测定法

• 批准号: 11557018
• 负责人: MORIUCHI Tetsuya
• 金额: $ 4.42万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11557018/
• 关键词: yeast; genetic diagnosis; p53; APC; PTEN; AIE-75; ADE2

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.

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。

项目成果

守内哲也: "Distinct prognostic values of p53 mutation and loss of estrogen receptor and their cumulative effect in primary breast cancers"Int.J.Cancer. 89. 92-99 (2000)

Tetsuya Moriuchi：“p53 突变和雌激素受体缺失的独特预后价值及其在原发性乳腺癌中的累积效应”Int.J.Cancer 89. 92-99 (2000)

Furuuchi, K.: "Somatic mutations of the APC gene in primary breast cancers"Am.J.Pathol.. 156. 1997-2005 (2000)

Furuuchi, K.：“原发性乳腺癌中 APC 基因的体细胞突变”Am.J.Pathol.. 156. 1997-2005 (2000)

Kobayashi,I.: "Identification of an autoimmune enteropathy-related 75-kilodalton antigen"Gastroenterology. 117. 823-830 (1997)

Kobayashi,I.：“自身免疫性肠病相关 75 千道尔顿抗原的鉴定”胃肠病学。

Zhang CL, Tada M, Kobayashi H, Nozaki M, Moriuchi T, Abe H.: "Detection of PTEN nonsense mutation and psiPTEN expression in central nervous system high-grade astrocytic tumors by a yeast-based stop codon assay."Oncogene.. 19. 4346-53 (2000)

张 CL、Tada M、Kobayashi H、Nozaki M、Moriuchi T、Abe H.：“通过基于酵母的终止密码子测定检测中枢神经系统高级星形细胞肿瘤中的 PTEN 无义突变和 psiPTEN 表达。”Oncogene..

Furuuchi K: "Somatic mutations of the APC gene in primary breast cancers"Am. J. Pathol. 156. 1997-2005 (2000)

Furuuchi K：“原发性乳腺癌中 APC 基因的体细胞突变”Am。