Conjugation in Escherichia coli: a study of recombination and the fate of donor DNA at the level of the zygote.
We have developed an experimental system for studying concomitantly the fate of the donor DNA and the process of recombination after conjugation in Escherichia coli. We used a set of Hfr and F-strains carrying complementing lacZ mutations. Expression of the lacZ allele on the chromosomal fragment derived from the donor results in the formation of heat sensitive beta-galactosidase by complementation. By intragenic recombination between the two lacZ mutations a lacZ+ gene may be formed, and wild type beta-galactosidase will be synthesized subsequently. So the assay of heat sensitive and wild type beta-galactosidase enabled us to follow respectively the fate of the donor DNA and the recombination process. Using various recombination deficient recipient strains, we found that the donor DNA is progressively inactivated in recA, rec-34 and recH recipients, although the initial rate of expression is equivalent to that in a Rec+ recipient; no significant recombination was observed. In Rec+, recB or recG recipients there was no inactivation and recombination occurred. The kinetics of recombinant formation in the recB strain seems to differ from the wild type; in a recG recipient the recombination activity is significant, but lower than in the wild type recipient.
D003227:Conjugation, Genetic; D004269:DNA, Bacterial / Q000032:analysis*; D004926:Escherichia coli / Q000201:enzymology / Q000502:physiology; D005260:Female; D005696:Galactosidases / Q000032:analysis; D009154:Mutation; D011995:Recombination, Genetic; D015053:Zygote
D004269:DNA, Bacterial; D005696:Galactosidases