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Reduction of marker discrimination in transductional recombination
Authors:Millicent Masters  Barbara J Newman and Christine M Henry
Institution:(1) Department of Molecular Biology, University of Edinburgh, King's Buildings, Mayfield Road, EH9 3JR Edinburgh, U.K.;(2) Present address: Dept. of Molecular Biochemistry and Biophysics, Yale University School of Medicine, 3333 Cedar Street, New Haven, Connecticut, USA
Abstract:Summary The recovery of phage P1 mediated transductants varies with the marker selected in a manner which cannot be fully accounted for by dosage differences in the donor gene population. This variation in transduction frequency is due primarily to recombinational discrimination in the recipient cell. We show here that increasing the intracellular level of recA protein, which might be expected to increase the contribution of recF mediated events to recombinant formation, decreases this discrimination slightly, and that replacing recBC mediated recombination by a recF dependent process, augmented by an additional, as yet uncharacterized mutation, dramatically reduces recombinational discrimination. We conclude that although recBC mediated transductional recombination is selective, recombination which relies on recF need not be so. We also show that UV-damaged DNA can be successfully recombined in the absence of the recB product (even in sbcB * cells) and that eliminating exonuclease I (the sbcB product) facilitates the recombination of heavily irradiated DNA.
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