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Titolo:
DEVELOPMENT OF A NOVEL CHL IU CELL-LINE WITH AN INCORPORATED GPT SHUTTLE VECTOR FOR CONCURRENT ANALYSIS OF GENE-MUTATIONS AND CHROMOSOME-ABERRATIONS/
Autore:
YAMADA T; ODAWARA K; KISHIDA F; NAKATSUKA I; YOSHITAKE A;
Indirizzi:
SUMITOMO CHEM CO LTD,ENVIRONMENTAL & HLTH SCI LAB,3 CHOME,KASUGADE NAKA KONOHANA OSAKA 554 JAPAN
Titolo Testata:
Mutation research. Genetic toxicology and environmental mutagenesis
fascicolo: 3, volume: 393, anno: 1997,
pagine: 295 - 306
SICI:
1383-5718(1997)393:3<295:DOANCI>2.0.ZU;2-7
Fonte:
ISI
Lingua:
ENG
Soggetto:
HYPOXANTHINE-GUANINE PHOSPHORIBOSYLTRANSFERASE; DIPLOID HUMAN FIBROBLASTS; DNA-SEQUENCE CHANGES; HAMSTER OVARY CELLS; ESCHERICHIA-COLI; MAMMALIAN-CELLS; ETHYL METHANESULFONATE; MOLECULAR ANALYSES; COSMID VECTORS; AS52 CELLS;
Keywords:
SHUTTLE VECTOR; GPT; CHL/IU CELL LINE;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Science Citation Index Expanded
Citazioni:
43
Recensione:
Indirizzi per estratti:
Citazione:
T. Yamada et al., "DEVELOPMENT OF A NOVEL CHL IU CELL-LINE WITH AN INCORPORATED GPT SHUTTLE VECTOR FOR CONCURRENT ANALYSIS OF GENE-MUTATIONS AND CHROMOSOME-ABERRATIONS/", Mutation research. Genetic toxicology and environmental mutagenesis, 393(3), 1997, pp. 295-306

Abstract

A cosmid shuttle vector containing the target gene of Escherichia coli gpt coding xanthine-guanine phosphoribosyl transferase was constructed. The shuttle vector was designed to be rescued into the gpt-deficient Escherichia coli from Chinese hamster CHL/IU cells through an in vitro packaging method. Mutations occurred at the target gene can be detected with a selective agent, 6-thioguanine (6-TG). The shuttle vectorwas stably transfected into CHL/IU cells to give several cell lines containing copies of the shuttle vector in the chromosomes. Each cell line exhibited a characteristic rescue efficiency (0 to 1.9 x 10(5) CFU/mu g of genomic DNA) of the shuttle vector and spontaneous mutation frequency (3.9 x 10(-5) to over 10(-2)) at the 6-TG selection. One transgenic cell line (KN63), which showed a higher rescue efficiency and alow spontaneous mutation frequency, was selected and tested for the ability to respond to a genotoxic agent, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). MNNG increased both the mutation frequency at the target gene and the number of the cells with chromosome aberrations. DNA sequence analysis of 6-TG mutants showed that predominant mutations (10/14) were identified as G:C to A:T transitions in MNNG-induced mutants,whereas transversions were predominant (5/9) in spontaneous mutants. These results suggest that this transgenic CHL/IU cell line can be a useful tool for analyzing the relation between gene mutations and chromosome aberrations. (C) 1997 Elsevier Science B.V.

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Documento generato il 21/09/20 alle ore 02:36:09