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Characterization of c-Ki-ras oncogene alleles by direct sequencing of enzymatically amplified DNA from carcinogen-induced tumors

Journal Article · · Proc. Natl. Acad. Sci. U.S.A.; (United States)
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Activated c-Ki-ras genes in liver tumors from rats exposed to the potent hepatocarcinogen aflatoxin B/sub 1/ were analyzed to determine the nature of their activation by characterization of two c-Ki-ras alleles present in tumor-derived NIH 3T3 mouse transformants. Using selective hybridization of synthetic oligonucleotides to transformant DNA, the authors have determined that a single G x C to A x T base transition in either the first or second position of the 12th codon is associated with activation of the gene. Such mutations would lead to amino acid substitutions of aspartate or serine for glycine in the mutant proteins. To confirm these findings, they applied a technique for direct sequence analysis of a 90-base-pair region of the rat c-Ki-ras gene produced by primer-directed enzymatic amplification. Findings produced by this approach, which provides a convenient method to characterize mutations in multiple alleles without the necessity to clone individual genes, confirmed the presence and identify of the 12th codon mutations in the activated oncogene, as initially determined by the oligonucleotide hybridization technique.

Research Organization:
Massachusetts Institute of Technology, Cambridge (USA)
OSTI ID:
5284020
Journal Information:
Proc. Natl. Acad. Sci. U.S.A.; (United States), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Vol. 84:14; ISSN PNASA
Country of Publication:
United States
Language:
English

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Related Subjects

550201 -- Biochemistry-- Tracer Techniques
560300* -- Chemicals Metabolism & Toxicology
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
AFLATOXIN
ANIMALS
ANTIGENS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL EFFECTS
BODY
CODONS
DAYS LIVING RADIOISOTOPES
DIGESTIVE SYSTEM
DISEASES
DNA SEQUENCING
GENE AMPLIFICATION
GENE MUTATIONS
GENES
GLANDS
HYBRIDIZATION
ISOTOPES
LIGHT NUCLEI
LIVER
MAMMALS
MATERIALS
MUTATIONS
NEOPLASMS
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
OLIGONUCLEOTIDES
ONCOGENES
ORGANIC COMPOUNDS
ORGANS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
RADIOISOTOPES
RATS
RODENTS
STRUCTURAL CHEMICAL ANALYSIS
TOXIC MATERIALS
TOXINS
VERTEBRATES