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A method to detect and characterize point mutations in transcribed genes: amplification and overexpression of the mutant cKi-ras allele in human tumor cells

AuthorsWinter, E.; Yamamoto, Fumiko; Almoguera, Concepción CSIC ORCID ; Perucho, M.
KeywordsHuman tumors
Activated ras oncogenes
C-ki-ras genes
Tumor cells
Issue Date1985
PublisherNational Academy of Sciences (U.S.)
CitationProceedings of the National Academy of Sciences of the USA Vol. 82, Issue 22, 7575-7579, (1985)
AbstractA 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 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.
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