Quantitative and molecular analyses of mutation in a pSV2gpt transformed CHO cell line
Following NDA-mediated gene transfer we have isolated a cell line useful for studying gene mutation at the molecular level. This line, AS52, derived from a hypoxanthine-guanine phosphoribosyl transferase (HGPRT) deficient Chinese hamster ovary (CHO) cell line, carries a single copy of the E. coli xanthine-guanine phosphoribosyl transferase (XGPRT) gene (gpt) and exhibits a spontaneous mutant frequency of 20 TG/sup r/ mutants/10/sup 6/ clonable cells. As with HGPRT/sup -/ mutants, XGPRT/sup -/ mutants can be selected in 6-thioguanine. AS52 (XGPRT/sup +/) and wild type CHO (HGPRT/sup +/) cell exhibit almost identical cytotoxic responses to various agents. We observed significant differences in mutation induction by UV light and ethyl methanesulfonate (EMS). Ratios of XGPRT/sup -/ to HGPRT/sup -/ mutants induced per unit dose (J/m/sup 2/ for UV light and ..mu..g/ml for EMS) are 1.4 and 0.70, respectively. Preliminary Southern blot hybridization analyses has been performed on 30 XGPRT/sup -/ AS52 mutants. A majority of spontaneous mutants have deletions ranging in size from 1 to 4 kilobases (9/19) to complete loss of gpt sequences (4/19); the remainder have no detectable (5/19) or only minor (1/19) alterations. 5/5 UV-induced and 5/6 EMS-induced mutants do not show a detectable change. Similar analyses are underway for mutations induced by x-irradiation and ICR 191 treatment.
- Research Organization:
- Tennessee Univ., Oak Ridge (USA). Graduate School of Biomedical Sciences; Oak Ridge National Lab., TN (USA)
- DOE Contract Number:
- W-7405-ENG-26
- OSTI ID:
- 5404031
- Report Number(s):
- CONF-831258-1; ON: DE84004428
- Resource Relation:
- Conference: 15. international congress of genetics, satellite symposisum on mutagenesis: basic and applied, Darbhanga, India, 22 Dec 1983
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
CHO CELLS
MUTANTS
DNA
ELECTROPHORESIS
EMS
MUTAGENESIS
GENES
RECOMBINANT DNA
MOLECULAR BIOLOGY
NUCLEOTIDYLTRANSFERASES
ENZYME ACTIVITY
ULTRAVIOLET RADIATION
BIOLOGICAL RADIATION EFFECTS
CELL CULTURES
ESCHERICHIA COLI
PLASMIDS
ANIMAL CELLS
BACTERIA
BIOLOGICAL EFFECTS
CELL CONSTITUENTS
ELECTROMAGNETIC RADIATION
ENZYMES
ESTERS
MICROORGANISMS
MUTAGENS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
RADIATION EFFECTS
RADIATIONS
SULFONIC ACID ESTERS
TRANSFERASES
560121* - Radiation Effects on Cells- External Source- (-1987)
560301 - Chemicals Metabolism & Toxicology- Cells- (-1987)