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RG. 3. Identification of 8-oxoG-induced mutations by restriction enzyme analysis. Form I DNA isolated from individual bacterial colonies was digested with NgoMYV and the products were analyzed on 1% (w/v) agarose gels. The original shuttle vector plasmid pS 189 has one JVgoMIV site and another is present in the inserted 8-oxoG-containing oligonucleotide. Thus the plasmids harboring a wild-type G : C sequence at the site of the original GO : C contain two restriction sites, giving rise to two DNA fragments of3407 and 1930 bp, respectively (lanes 2-7), whereas plasmids mutated at the site originally containing the 8-oxoG are cut only once (lanes 1 and 8). The use of this enzyme therefore allows us to quantify the mutation frequency. All mutated plasmids were confirmed by DNA sequencing: (-) undigested DNA; (+) iVgoMIV-digested DNA plasmids.

RG. 3. Identification of 8-oxoG-induced mutations by restriction enzyme analysis. Form I DNA isolated from individual bacterial colonies was digested with NgoMYV and the products were analyzed on 1% (w/v) agarose gels. The original shuttle vector plasmid pS 189 has one JVgoMIV site and another is present in the inserted 8-oxoG-containing oligonucleotide. Thus the plasmids harboring a wild-type G : C sequence at the site of the original GO : C contain two restriction sites, giving rise to two DNA fragments of3407 and 1930 bp, respectively (lanes 2-7), whereas plasmids mutated at the site originally containing the 8-oxoG are cut only once (lanes 1 and 8). The use of this enzyme therefore allows us to quantify the mutation frequency. All mutated plasmids were confirmed by DNA sequencing: (-) undigested DNA; (+) iVgoMIV-digested DNA plasmids.

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