Premium Efficient detection of deletions induced by a single treatment of mitomycin C in transgenic mouse gpt delta using the Spi − selection
Okada Naoko,
Masumura Kenichi,
Nohmi Takehiko,
Yajima Nobuhiro
Publication year1999
Publication title
environmental and molecular mutagenesis
Resource typeJournals
Abstract Transgenic rodent mutation assays permit the detection and molecular analysis of various types of gene mutations, such as base changes and frameshifts, in a number of tissues. It is reported, however, that deletion mutations are not efficiently detected by the assays, in particular those using lambda phage shuttle vectors. Recently, a new transgenic mouse model, i.e., gpt delta, has been developed to selectively detect some types of deletions by Spi − selection. Spi − selection has an advantage over the other selections to preferentially identify deletions because only lambda phages deficient in both the red and gam gene functions are allowed to form phage plaques. In this study, we examined whether in vivo deletions induced by the treatment of mitomycin C (MMC) are detectable by the Spi − assay in the mouse model. The mice were treated with MMC (0.5, 1.0, 2.0, and 4.0 mg/kg, single intraperitoneal injection) and sacrificed 14 days after the dosing. The treatment at 4.0 mg/kg approximately doubled the mutant frequency of Spi − in the bone marrow, i.e., 2.52 × 10 −6 vs. 1.31 × 10 −6 . The molecular analyses using polymerase chain reaction (PCR) and DNA sequencing indicated that seven Spi − mutants at 4.0 mg/kg group had deletions with molecular sizes from 0.8 kilo basepairs (kb) to 8.5 kb, whereas no such deletions were observed in the Spi − mutants in the control group. The results suggest that deletions induced by MMC in the bone marrow are efficiently detectable by Spi − selection and also that the molecular analyses are useful to evaluate the significance of a marginal increase in mutant frequency in the transgenic rodent mutation assays. Environ. Mol. Mutagen. 34:106–111, 1999 © 1999 Wiley‐Liss, Inc.
Subject(s)biology , dna , gene , genetically modified mouse , genetics , microbiology and biotechnology , mitomycin c , mutagen , mutagenesis , mutant , mutation , polymerase chain reaction , transgene
SCImago Journal Rank1

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