Site-specific mutagenesis in cells with normal DNA repair systems: transitions produced from DNA carrying a single O6-alkylguanine
Open Access
- 1 January 1991
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 19 (9) , 2485-2488
- https://doi.org/10.1093/nar/19.9.2485
Abstract
This paper describes a systematic study of transition frequencies produced in vivo when a homologous series of O6-alkylguanine residues located at a preselected position in gene G of øX174 form I' DNA (double-stranded, circular, covalently-closed, relaxed) Is transfected into spheroplasts from two strains of Escherichla coli having normal DNA repair systems. Mutant frequencies were measured as percent of total phage produced by single bursts. The results are: (A) Synthetic DNA without any alkyl group gave a transition frequency of 0.02%. (B) In E. coli AB1157, the frequencies fall into two groups depending on the alkyl group: methyl and ethyl, 8 – 1%; n-propyl and n-butyl ∼0.9%. (C) The average transition frequencies were higher in AB1157 than in C600. These data demonstrate that a single O6-alkylguanine residue can produce a specific transition at significant frequencies in cells with normal repair systems and that the mutant frequency depends upon the nature of the alkyl group and the cell type.Keywords
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