Initiation of chromosome replication in Escherichia coli after induction of dnaA gene expression from a lac promoter.

Abstract

Escherichia coli HB282 carries a dnaA46(Ts) allele on the chromosome, a wild-type dnaA allele under the control of the lacUV5 promoter on the multicopy plasmid pBC32, and an overproducing lac repressor allele on an F' factor. When the plasmid dnaA gene is repressed, the strain is thermosensitive. After a temporary deficiency in active dnaA protein at nonpermissive temperature, the addition of isopropyl-beta-D-thiogalactopyranoside to the culture was found to produce a burst of initiations within 5 to 10 min at 30% of the origins in 90% of the cells. Initiations then continued at a rate slightly faster than the mass-doubling time such that after 2 h the origin-to-mass ratio of the control culture was restored.