Sensitive SV40 viral probe assay for DNA strand breaks and their biological repair in higher cells: techniques and preliminary results
An alternative experimental approach to the investigation of intracellular repair of DNA strand breaks by mammalian higher cells has been developed using simian virus 40 (SV40), which has no known intrinsic DNA repair capacity and possesses a minichromosome structure, as an intracellular probe. In this approach unirradiated simian or human cells are infected with irradiated virus and incubated for varying periods. This assay involves separation of the viral DNA by agarose gel electrophoresis into three sharply demarcated bands corresponding to DNA molecules containing a double-strand break (DSB), single-strand breaks (SSB), or no breaks. Quantitative data are obtained by a combination of DNA hybridization with /sup 32/P-labeled SV40 DNA, autoradiography, and densitometry. These experiments indicate that there is rapid and efficient cellular uptake of SV40, independent of prior radiation dose to the virus, and that this multistep experimental procedure gives excellent recovery and quantitation of the three DNA forms when compared with more direct methods of measurement. Initial time-course incubation experiments with SV40 infected CV-1 monkey kidney cells indicate that this approach can demonstrate slow but extensive intracellular repair of SSB; and limited presumptive early repair of DSB, followed by later and more extensive induction of DSB.
- Research Organization:
- Univ. of Pennsylvania School of Medicine, Philadelphia
- DOE Contract Number:
- AC02-83ER60163
- OSTI ID:
- 5779651
- Journal Information:
- Radiat. Res.; (United States), Journal Name: Radiat. Res.; (United States) Vol. 101:2; ISSN RAREA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
560131 -- Radiation Effects on Microorganisms-- Basic Studies-- (-1987)
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ALKALI METAL ISOTOPES
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOASSAY
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
CESIUM 137
CESIUM ISOTOPES
DAYS LIVING RADIOISOTOPES
DNA
ELECTROMAGNETIC RADIATION
ELECTROPHORESIS
GAMMA RADIATION
IONIZING RADIATIONS
ISOTOPES
LABELLED COMPOUNDS
LIGHT NUCLEI
MICROORGANISMS
NUCLEI
NUCLEIC ACIDS
ODD-EVEN NUCLEI
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PARASITES
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
RADIATIONS
RADIOISOTOPES
RECOVERY
REPAIR
SIMIAN VIRUS
STRAND BREAKS
TRITIUM COMPOUNDS
VIRUSES
YEARS LIVING RADIOISOTOPES