Factors affecting mutational specificity in mammalian cells: (Informal technical progress report), February 1, 1986-January 31, 1987
Technical Report
·
OSTI ID:7024799
We analyzed the different c-H-ras mutations produced in cells after treatment with chemical carcinogens. The overall goal of this work is an understanding of the changes produced by environmental mutagens and carcinogens to learn how closely the results obtained with bacterial and cellular assay systems apply to the in vivo situation. Our work continues to utilize the technique for the study of DNA synthesis termination in vitro developed in this laboraory and utilized for the past several years. This technology can be characterized as a Sanger dideoxy sequencing technique with mutation induced lesions in the template strand serving as chain terminators instead of the dideoxynucleotides used in the Sanger technique to terminate the growing strand. We have added to this technique a modification for the study of termination on double stranded templates as possibly modeling more closely the natural situation. For this modification, double stranded M13 DNA is split once with a restriction enzyme which makes a unique cut and the linearized DNA is hybridized with single stranded circular DNA to make a uniquely nicked double stranded circle with a single 3'OH to serve as a primer for DNA synthesis which can occur by either strand displacement or nick translation.
- Research Organization:
- Chicago Univ., IL (USA)
- DOE Contract Number:
- AC02-76EV02040
- OSTI ID:
- 7024799
- Report Number(s):
- DOE/EV/02040-T2; ON: DE88012387
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
550200 -- Biochemistry
560300* -- Chemicals Metabolism & Toxicology
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ACETYLAMINOFLUORENES
AMINES
BIOASSAY
CARCINOGENS
DNA BASE TRANSITIONS
DNA SEQUENCING
DOCUMENT TYPES
ENVIRONMENTAL EFFECTS
MUTAGENESIS
MUTAGENS
ORGANIC COMPOUNDS
POLYCYCLIC AROMATIC AMINES
PROGRESS REPORT
STRUCTURAL CHEMICAL ANALYSIS
560300* -- Chemicals Metabolism & Toxicology
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ACETYLAMINOFLUORENES
AMINES
BIOASSAY
CARCINOGENS
DNA BASE TRANSITIONS
DNA SEQUENCING
DOCUMENT TYPES
ENVIRONMENTAL EFFECTS
MUTAGENESIS
MUTAGENS
ORGANIC COMPOUNDS
POLYCYCLIC AROMATIC AMINES
PROGRESS REPORT
STRUCTURAL CHEMICAL ANALYSIS