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Methods Mol Biol 2013;1044:97-119

In vivo cII, gpt, and Spi┬┐ gene mutation assays in transgenic mice and rats.

Manjanatha MG, Cao X, Shelton SD, Mittelstaedt RA, Heflich RH

Abstract

Transgenic mutation assays are used to identify and characterize genotoxic hazards and for determining the mode of action for carcinogens. The three most popular transgenic mutational models are Big Blue((R)) (rats or mice), Muta mouse (mice), and gpt-delta (rats or mice). The Big Blue((R)) and Muta mouse models use the cII gene as a reporter of mutation whereas gpt-delta rodents use the gpt gene and the red/gam genes (Spi(-) selection) as mutation reporter genes. Here we describe methodology for conducting mutation assays with these transgenes. Transgenes recovered from tissue DNA are packaged into infectious lambda phage, bacteria are infected with the phage, and cII-mutant and Spi(-) plaques and gpt-mutant colonies are isolated using selective conditions and quantified. Selected mutants can be further analyzed for identification of small sequence alterations in the cII and gpt genes and large deletions at the Spi(-) locus.


Category: Journal Article
PubMed ID: #23896873 DOI: 10.1007/978-1-62703-529-3_5
Includes FDA Authors from Scientific Area(s): Toxicological Research
Entry Created: 2013-07-31 Entry Last Modified: 2014-02-25
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