Summary
Incubation of the colicinogenic Escherichia coli strain JC 411 (ColE1) at elevated temperatures (47–49°) leads to the accumulation of catenated molecules and replicative intermediates of this plasmid. Mature supercoiled ColE1 DNA molecules synthesized under these conditions have an increased number of tertiary turns as shown by electron microscopy. The monomeric tightly supercoiled molecules possess a slightly slower sedimentation rate and a higher binding capacity for ethidium bromide than supercoiled monomers synthesized at lower temperatures. Recombination deficient mutants of E. coli recA, recB and recC, which carry the ColE1 plasmid, form about the same amount of catenated molecules at the elevated temperature as a rec+ strain.
In addition, we have observed by electron microscopy a small percentage (∼5% of the circular DNA molecules) of minicircular DNA molecules in all preparations of JC 411 (ColE1). They are homogenous in size, with a molecular weight of 1.4x106 daltons.
Addition of chloramphenicol to a culture of Proteus mirabilis (ColE1) leads to an increased amount of higher multiple circular oligomers and to a stimulated accumulation of catenated ColE1 DNA molecules of varying sizes. ColE1 DNA synthesis is more thermosensitive than chromosomal DNA replication in P. mirabilis. Plasmid replication stops completely at temperatures above 43°C.
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Communicated by R. Devoret
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Goebel, W., Kreft, J. Complex ColE1 DNA in Escherichia coli and Proteus mirabilis . Molec. Gen. Genet. 129, 149–166 (1974). https://doi.org/10.1007/BF00268628
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DOI: https://doi.org/10.1007/BF00268628