Recombinant methods for screening human DNA excision repair proficiency
Thesis/Dissertation
·
OSTI ID:5513156
A method for measuring DNA excision repair in response to ultraviolet radiation (UV)-induced DNA damage has been developed, validated, and field-tested in cultured human lymphocytes. The methodology is amenable to population-based screening and should facilitate future epidemiologic studies seeking to investigate associations between excision repair proficiency and cancer susceptibility. The impetus for such endeavors derives from the belief that the high incidence of skin cancer in the genetic disorder xeroderma pigmentosum (XP) primarily is a result of the reduced capacity of patients cells to repair UV-induced DNA damage. For assay, UV-irradiated non-replicating recombinant plasmid DNA harboring a chloramphenicol acetyltransferase (CAT) indicator gene is introduced into lymphocytes using DEAE-dextran short-term transfection conditions. Exposure to UV induces transcriptionally-inactivating DNA photoproducts in the plasmid DNA which inactivate CAT gene expression. Excision repair of the damaged CAT gene is monitored indirectly as a function of reactivated CAT enzyme activity following a 40 hour repair/expression incubation period.
- Research Organization:
- Johns Hopkins Univ., Baltimore, MD (USA)
- OSTI ID:
- 5513156
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
560120* -- Radiation Effects on Biochemicals
Cells
& Tissue Culture
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANIMAL CELLS
ANIMALS
BIOLOGICAL MATERIALS
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
BLOOD
BLOOD CELLS
BODY FLUIDS
CHEMICAL RADIATION EFFECTS
CHEMISTRY
CONNECTIVE TISSUE CELLS
DISEASES
DNA
DNA REPAIR
ELECTROMAGNETIC RADIATION
ENZYME ACTIVITY
ENZYMES
GENE REGULATION
GENES
LEUKOCYTES
LYMPHOCYTES
MAMMALS
MAN
MATERIALS
MEASURING METHODS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PRIMATES
RADIATION CHEMISTRY
RADIATION EFFECTS
RADIATIONS
RECOMBINANT DNA
RECOVERY
REPAIR
SKIN DISEASES
SOMATIC CELLS
TRANSFERASES
ULTRAVIOLET RADIATION
VERTEBRATES
XERODERMA PIGMENTOSUM
Cells
& Tissue Culture
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANIMAL CELLS
ANIMALS
BIOLOGICAL MATERIALS
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
BLOOD
BLOOD CELLS
BODY FLUIDS
CHEMICAL RADIATION EFFECTS
CHEMISTRY
CONNECTIVE TISSUE CELLS
DISEASES
DNA
DNA REPAIR
ELECTROMAGNETIC RADIATION
ENZYME ACTIVITY
ENZYMES
GENE REGULATION
GENES
LEUKOCYTES
LYMPHOCYTES
MAMMALS
MAN
MATERIALS
MEASURING METHODS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PRIMATES
RADIATION CHEMISTRY
RADIATION EFFECTS
RADIATIONS
RECOMBINANT DNA
RECOVERY
REPAIR
SKIN DISEASES
SOMATIC CELLS
TRANSFERASES
ULTRAVIOLET RADIATION
VERTEBRATES
XERODERMA PIGMENTOSUM