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Detection of deletion mutations in pSV2gpt-transformed cells

Journal Article · · Mol. Cell. Biol.; (United States)
DOI:https://doi.org/10.1128/MCB.4.7.1411· OSTI ID:5072134
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The authors have developed a system to study mutations that affect xanthine-guanine phosphoribosyltransferase gene (gpt) expression in hypoxanthine-guanine phosphoribosyltransferase-deficient CHO cells that have been transformed by the plasmid vector pSV2gpt. One isolated transformant, designated AS52, carries a single copy of the Escherichia coli gpt gene stably integrated into the high-molecular-weight DNA and expresses the bacterial gene for the enzyme xanthine-guanine phosphoribosyltransferase. Mutants deficient in this enzyme can be induced in the AS52 cell line by a variety of mutagens, and spontaneous or induced mutants can be selected for resistance to 6-thioguanine (Tg/sup r/). Two Tg/sup r/ clones derived from the AS52 line were analyzed by Southern blot hybridization and were found to contain deletions involving at least a portion of the gpt gene. Because of the small size and stability of the integrated pSV2gpt plasmid, and the well-defined selection protocol for mutant isolation, the AS52 line offers promise as a system suitable for the study of mutation at the molecular level in CHO cells.
Research Organization:
Oak Ridge National Lab., TN
DOE Contract Number:
W-7405-ENG-26
OSTI ID:
5072134
Journal Information:
Mol. Cell. Biol.; (United States), Journal Name: Mol. Cell. Biol.; (United States) Vol. 4:7; ISSN MCEBD
Country of Publication:
United States
Language:
English

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Related Subjects

550400 -- Genetics
560121* -- Radiation Effects on Cells-- External Source-- (-1987)
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANIMAL CELLS
BIOLOGICAL EFFECTS
BIOLOGICAL RADIATION EFFECTS
BIOLOGY
CHO CELLS
ELECTROMAGNETIC RADIATION
ENZYME ACTIVITY
ENZYMES
EXTREME ULTRAVIOLET RADIATION
GENES
GENETIC EFFECTS
GENETIC RADIATION EFFECTS
GENETICS
GLYCOSYL TRANSFERASES
HYPOXANTHINE PHOSPHORIBOSYLTRANSFERASE
IONIZING RADIATIONS
MICROORGANISMS
MUTATIONS
PARASITES
PENTOSYL TRANSFERASES
RADIATION EFFECTS
RADIATIONS
SIMIAN VIRUS
TRANSFERASES
ULTRAVIOLET RADIATION
VIRUSES
X RADIATION