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Loss of Hfr DNA from Escherichia coli merozygotes during inhibition of conjugation by nalidixic acid

Published online by Cambridge University Press:  14 April 2009

Robert G. Lloyd ,
Janet Hart  and
Sandra Johnson
Robert G. Lloyd
Affiliation:
Department of Genetics, University of Nottingham, Nottingham NG7 2RD, England
Janet Hart
Affiliation:
Department of Genetics, University of Nottingham, Nottingham NG7 2RD, England
Sandra Johnson
Affiliation:
Department of Genetics, University of Nottingham, Nottingham NG7 2RD, England
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The effect of nalidixic acid on conjugal recombination was studied in matings with recipient strains carrying recA200, a mutation which confers a thermosensitive Rec− phenotype. Addition of nalidixic acid to Hfr Nals × F−NalRrecA200 matings at low temperature (35 °C) caused a sharp 10- to 20-fold decline in the yield of recombinants if plating on selective agar was delayed. Two separate processes were identified as being responsible for this decline. Those merozygotes in which the transferred DNA was free of the donor cell lost the ability to form recombinants through inactivation of this DNA, an effect which could be prevented by using exonuclease deficient (recB sbcB) recipients or by prior growth of exonuclease proficient recipients in medium containing 0·25 M sodium chloride. No more than 50% of the observed loss of recombinants could be attributed to this effect. The remaining merozygotes lost their ability for recombinant formation provided mating pairs, and presumably the displaced donor DNA strand, remained intact. This process was thought to involve withdrawal of transferred DNA (DeHaan & Gross, 1962) and was studied in isolation in matings with recA+, or recA200 recB sbcB recipients. A mechanism involving re-annealing of the displaced Hfr DNA to the donor molecule as a result of nalidixic inhibition of gyrase activity in the donor causing relaxation of DNA supercoils is proposed to account for this withdrawal event.

Type
Research Article
Information
Genetics Research , Volume 36 , Issue 1 , August 1980 , pp. 69 - 79
Copyright
Copyright © Cambridge University Press 1980

References

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