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Titolo:
A NEW TRANSGENIC MOUSE MUTAGENESIS TEST SYSTEM USING SPI(-) AND 6-THIOGUANINE SELECTIONS
Autore:
NOHMI T; KATOH M; SUZUKI H; MATSUI M; YAMADA M; WATANABE M; SUZUKI M; HORIYA N; UEDA O; SHIBUYA T; IKEDA H; SOFUNI T;
Indirizzi:
NATL INST HLTH SCI,DIV GENET & MUTAGENESIS,SETAGAYA KU,1-18-1 KAMIYOGA TOKYO 158 JAPAN UNIV CHILE,FAC ODONTOL SANTIAGO CHILE CHUGAI PHARMACEUT CO LTD,FUJI GOTEMBA RES LAB,EXPLORATORY RES LAB GOTEMBA SHIZUOKA JAPAN NATL CANC CTR,RES INST,DIV BIOCHEM,CHUO KU TOKYO 104 JAPAN HATANO RES INST,LAB GENET TOXICOL KANAGAWA JAPAN UNIV TOKYO,INST MED SCI,DEPT BIOL MOL TOKYO JAPAN
Titolo Testata:
Environmental and molecular mutagenesis
fascicolo: 4, volume: 28, anno: 1996,
pagine: 465 - 470
SICI:
0893-6692(1996)28:4<465:ANTMMT>2.0.ZU;2-V
Fonte:
ISI
Lingua:
ENG
Soggetto:
HAMSTER OVARY CELLS; ESCHERICHIA-COLI; ULTRAVIOLET-LIGHT; GPT GENE; ILLEGITIMATE RECOMBINATION; MOLECULAR ANALYSIS; MUTATIONS INVIVO; SEQUENCE CHANGES; AS52 CELLS; X-RAY;
Keywords:
TRANSGENIC MOUSE; GPT; SPI SELECTION; POINT MUTATION; DELETION MUTATION; GAMMA-RAY;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Science Citation Index Expanded
Citazioni:
29
Recensione:
Indirizzi per estratti:
Citazione:
T. Nohmi et al., "A NEW TRANSGENIC MOUSE MUTAGENESIS TEST SYSTEM USING SPI(-) AND 6-THIOGUANINE SELECTIONS", Environmental and molecular mutagenesis, 28(4), 1996, pp. 465-470

Abstract

A new transgenic mouse mutagenesis test system has been developed Forthe efficient detection of point mutations and deletion mutations in vivo. The mice carry lambda EG10 DNA as a transgene. When the rescued phages are infected into Escherichia coli YG6020-expressing Cre recombinase, the phage DNA is converted into plasmid pYG142 carrying the chloramphenicol-resistance gene and the gpi gene of E. coli. The gpt mutants can be positively detected as colonies arising on plates containing chloramphenicol and 6-throguanine. The EG10 DNA carries a chi site along with the red and gam genes so that the wild-type phages display Spi(+) (sensitive to P2 interference) phenotype. Mutant phages lacking both red and gam genes can be positively detected as plaques that growin P2 lysogens of E. coli. These mutant phages are called lambda Spi(-). The spontaneous gpi mutation frequencies of five independent transgenic lines were 1.7 to 3.3 x 10(-5) in bone marrow. When the mice were treated with ethylnitrosourea (single i.p. treatments with 150 mg/kgbody weight; killed 7 days after the treatments), mutation frequencies were increased four- to sevenfold over the background in bone marrow. The average rescue efficiencies were more than 200,000 chloramphenicol-resistant colonies per 7.5 mu g bone marrow DNA per packaging reaction. In contrast to gpi mutation frequencies, spontaneous Spi(-) mutation Frequencies were 1.4 x 10(-6) and 1.1 x 10(-6) in bone marrow and sperm, respectively. No spontaneous Spi(-) mutants have been detected so far in spleen, although 930,000 phages rescued from untreated mice were screened. In gamma-ray-treated animals, however, induction of Spi(-) mutations was clearly observed in spleen, at frequencies of 1.4 x 10(-5) (5 Gy), 1.2 x 10(-5) (10 GY), and 2.0 x 10(-5) (50 Gy). These results suggest that the new transgenic mouse ''gpt delta'' could be useful for the efficient detection of point mutations and deletion mutations in vivo. (C) 1996 Wiley-Liss, Inc.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 21/09/20 alle ore 03:35:08