Detection of deletion mutations in pSV2gpt-transformed cells
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. (ORNL), Oak Ridge, TN (United States)
- DOE Contract Number:
- W-7405-ENG-26
- OSTI ID:
- 5072134
- Journal Information:
- Mol. Cell. Biol.; (United States), Vol. 4:7
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
CHO CELLS
ENZYME ACTIVITY
GENETIC RADIATION EFFECTS
HYPOXANTHINE PHOSPHORIBOSYLTRANSFERASE
GENETICS
EXTREME ULTRAVIOLET RADIATION
GENES
MUTATIONS
SIMIAN VIRUS
X RADIATION
ANIMAL CELLS
BIOLOGICAL EFFECTS
BIOLOGICAL RADIATION EFFECTS
BIOLOGY
ELECTROMAGNETIC RADIATION
ENZYMES
GENETIC EFFECTS
GLYCOSYL TRANSFERASES
IONIZING RADIATIONS
MICROORGANISMS
PARASITES
PENTOSYL TRANSFERASES
RADIATION EFFECTS
RADIATIONS
TRANSFERASES
ULTRAVIOLET RADIATION
VIRUSES
560121* - Radiation Effects on Cells- External Source- (-1987)
550400 - Genetics