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System issues: The genetic analysis of lacl mutations in sectored plaques from Big Blue® transgenic mice

✍ Scribed by Gregory R. Stuart; Nancy J. Gorelick; Julie L. Andrews; Johan G. de Boer; Barry W. Glickman

Publisher
John Wiley and Sons
Year
1996
Tongue
English
Weight
761 KB
Volume
28
Category
Article
ISSN
0893-6692

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✦ Synopsis

However, the interpretation of data from transgenic animal assays is sometimes complicated by mutants that appear as sectored mutant lambda plaques. These mutants can form a significant fraction of the mutant plaques [Hayward et al. (1 995): Carcinogenesis 16:2429-24331. Thus, in order to accurately determine in vivo mutant frequencies and mutational specificities, it is necessary to score sectored plaques and partition them from the rest of the data. In this study, the specificity of mutation in sectored plaques recovered from untreated and UVB-treated Big Blue@ mouse skin was analyzed and compared to mutations recovered from hLlZ//ac/ grown on the Escherichia coli host. The mutational spectra of sectored plaques from untreated and UVB-treated mice were remarkably similar to each other and resembled those recovered from the XLIZ/lac/ phage plated directly on E. coli. Both the sectored mutants and those recovered in hLlZ//ac/ phage differed from the spectra of spontaneous mutants in E.

coli and in Big Blue@ mouse skin. While sectored mutants from UVB-treated mouse skin and XLIZ//ac/ mutants were also different from spontaneous mutants recovered from Big Blue@ liver, there was little difference between sectored mutants from untreated mouse skin and spontaneous liver mutants (P = 0.07). The mutational spectra of sectored plaques is thus largely consistent with their origin as spontaneous mutations arising in vitro during growth of the hLlZ/lac/ shuttle vector DNA on the E. coli host, although the potential contribution from lesions in mouse DNA being expressed ex vivo in the E. coli host cannot be excluded.

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