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

Winter, E.; Yamamoto, Fumiko; Almoguera, Concepción; Perucho, M.; National Cancer Institute; Asahi Kasei Corporation

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

1985

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.

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This work was supported by Grants CA33021 and CA38579 awarded by the National Cancer Institute and by a grant from Toyo Jozo Co. Ltd.

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Peer reviewed

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