Progress in identifying a human ionizing-radiation repair gene using DNA-mediated gene transfer techniques

Conference · Thu May 01 04:00:00 EDT 1986 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:5204109

Chen, D J.C.; Strniste, G F; Jeggo, P A

The authors employing DNA-mediated gene transfer techniques in introducing human DNA into a DNA double-strand break (DSB) repair deficient Chinese hamster (CHO) cell mutant (xrs-6), which is hypersensitive to both X-rays (D/sub 0/ = 0.39 Gy) and the antibiotic bleomycin (D/sub 0/ = 0.01 ..mu..g/ml). High molecular weight DNA isolated from cultured human skin fibroblasts was partially digested with restriction enzyme Sau 3A to average sizes of 20 or 40 Kb, ligated with plasmid pSV2-gpt DNA, and transfected into xrs-6 cells. Colonies which developed under a bleomycin and MAX (mycophenolic acid/adenine/xanthine) double-selection procedure were isolated and further tested for X-ray sensitivity and DSB rejoining capacity. To date a total of six X-ray or bleomycin resistant transformants have been isolated. All express rejoining capacity for X-ray-induced DSB, similar to the rate observed for DSB repair in CHO wild type cells. DNA isolated from these primary transformants contain various copy numbers of pSV2-gpt DNA and also contain human DNA sequences as determined by Southern blot hybridization. Recently, a secondary transformant has been isolated using DNA from one of the primary transformants. Cellular and molecular characterization of this transformant is in progress. DNA from a genuine secondary transformant will be used in the construction of a DNA library to isolate human genomic DNA encoding this radiation repair gene.

• Research Organization: Los Alamos National Lab., NM
• OSTI ID: 5204109
• Report Number(s): CONF-8606151-
• Conference Information: Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 45:6
• Country of Publication: United States
• Language: English

Related Subjects

560121* -- Radiation Effects on Cells-- External Source-- (-1987)
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANIMAL CELLS
ANIMALS
ANTI-INFECTIVE AGENTS
ANTIBIOTICS
ANTIMITOTIC DRUGS
ANTINEOPLASTIC DRUGS
BIOLOGICAL EFFECTS
BIOLOGICAL RADIATION EFFECTS
BLEOMYCIN
CHO CELLS
DNA
DNA SEQUENCING
DRUGS
ELECTROMAGNETIC RADIATION
GENES
GENETIC EFFECTS
GENETIC RADIATION EFFECTS
IONIZING RADIATIONS
MAMMALS
MAN
MOLECULAR WEIGHT
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PRIMATES
RADIATION EFFECTS
RADIATIONS
RADIOSENSITIVITY
STRAND BREAKS
STRUCTURAL CHEMICAL ANALYSIS
VERTEBRATES
X RADIATION