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Use of a rapid DNA sequencing system to demonstrate the induction of frameshift mutations by bleomycin
Authors:N Demopoulos  R W Davies  C Scazzocchio
Institution:1. University of Patras, Laboratory of Genetics, Patras, Greece;2. University of Manchester Institute of Science and Technology, Applied Molecular Biology Group, Department of Biochemistry, PO Box 88, Manchester M60 1QD U.K.;3. University of Essex, Department of Biology, Colchester CO4 3SQ, England
Abstract:The rapid DNA sequencing system based on the single-stranded bacteriophage M13 and the chain-terminator method has been used to look directly for mutational alterations. A small DNA fragment that primes DNA synthesis through the N-terminal 200 base pairs of the beta-galactosidase gene was prepared, and used to detect changes in base sequence among phages that give white plaques after treatment of the host cells with bleomycin. Bleomycin treatment of E. coli in which M13 mp2 was growing gave an increase in white plaque frequency. DNA sequence analysis of phage from 7 independent mutant plaques showed them all to have a frameshift mutation.
Keywords:DNA sequencing M13  Mutagenesis screen  Bleomycin  Frameshift  Carcinogenesis
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