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Title: Quantitative and molecular analyses of mutation in a pSV2gpt transformed CHO cell line

Abstract

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 formore » mutations induced by x-irradiation and ICR 191 treatment.« less

Authors:
; ;
Publication Date:
Research Org.:
Tennessee Univ., Oak Ridge (USA). Graduate School of Biomedical Sciences; Oak Ridge National Lab., TN (USA)
OSTI Identifier:
5404031
Report Number(s):
CONF-831258-1
ON: DE84004428
DOE Contract Number:  
W-7405-ENG-26
Resource Type:
Conference
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
Subject:
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.; 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)

Citation Formats

Stankowski, Jr, L F, Tindall, K R, and Hsie, A W. Quantitative and molecular analyses of mutation in a pSV2gpt transformed CHO cell line. United States: N. p., 1983. Web.
Stankowski, Jr, L F, Tindall, K R, & Hsie, A W. Quantitative and molecular analyses of mutation in a pSV2gpt transformed CHO cell line. United States.
Stankowski, Jr, L F, Tindall, K R, and Hsie, A W. 1983. "Quantitative and molecular analyses of mutation in a pSV2gpt transformed CHO cell line". United States. https://www.osti.gov/servlets/purl/5404031.
@article{osti_5404031,
title = {Quantitative and molecular analyses of mutation in a pSV2gpt transformed CHO cell line},
author = {Stankowski, Jr, L F and Tindall, K R and Hsie, A W},
abstractNote = {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.},
doi = {},
url = {https://www.osti.gov/biblio/5404031}, journal = {},
number = ,
volume = ,
place = {United States},
year = {Sat Jan 01 00:00:00 EST 1983},
month = {Sat Jan 01 00:00:00 EST 1983}
}

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