A method to detect and characterize point mutations in transcribed genes: amplification and overexpression of the mutant c-Ki-ras allele in human tumor cells.
A method to detect and characterize point mutations in transcribed genes: amplification and overexpression of the mutant c-Ki-ras allele in human tumor cells.
复制标题
一种检测和表征转录基因点突变的方法：人类肿瘤细胞中突变 c-Ki-ras 等位基因的扩增和过度表达。
DOI：
10.1073/pnas.82.22.7575
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发表时间：
1985
影响因子：
11.1
通讯作者：
Perucho,M
中科院分区：
文献类型：
--
作者：
Winter,E;Yamamoto,F;Almoguera,C;Perucho,M
相当大比例的人类肿瘤含有激活的ras癌基因，其由于编码的ras基因产物的密码子12或61处的体细胞点突变而具有致癌潜力。我们在这里报告一种方法来检测和表征ras基因突变的基础上，胰腺核糖核酸酶（RNase A; EC 3.1.27.5）的能力，切割RNA异源双链含有单碱基错配。使用这种方法，我们表明，某些人类肿瘤细胞含有突变的c-Ki-ras基因，我们定义这些突变的性质和位置。在同一时间，我们描述的存在和估计的正常和突变的c-Ki-ras等位基因在相同的肿瘤细胞的表达。这种方法应该是有用的诊断检测和表达基因中的单点突变的特性。
A significant percentage of human tumors contain activated ras oncogenes that have acquired oncogenic potential as a result of somatic point mutations at codon 12 or 61 of the encoded ras gene product. We report here a method to detect and characterize mutations in ras genes that is based on the ability of pancreatic ribonuclease (RNase A; EC 3.1.27.5) to cleave RNA heteroduplexes containing single-base mismatches. Using this method, we show that certain human tumor cells contain mutant c-Ki-ras genes, and we define the nature and position of these mutations. At the same time, we describe the presence and estimate the expression of both normal and mutant c-Ki-ras alleles in the same tumor cells. This method should be useful for the diagnostic detection and characterization of single point mutations in expressed genes.