Winter E, Yamamoto F, Almoguera C, Perucho M
Proc Natl Acad Sci U S A. 1985 Nov;82(22):7575-9. doi: 10.1073/pnas.82.22.7575.
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.
相当大比例的人类肿瘤含有激活的ras癌基因，这些癌基因由于编码的ras基因产物第12或61密码子处的体细胞点突变而获得了致癌潜力。我们在此报告一种检测和鉴定ras基因突变的方法，该方法基于胰腺核糖核酸酶（RNase A；EC 3.1.27.5）切割含有单碱基错配的RNA异源双链体的能力。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，并确定了这些突变的性质和位置。同时，我们描述了同一肿瘤细胞中正常和突变c-Ki-ras等位基因的存在情况并估计了其表达水平。该方法对于诊断检测和鉴定表达基因中的单点突变应该是有用的。
