Trans-complementable copy-number mutants of plasmid ColE1 (original) (raw)
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- Published: 10 January 1980
Nature volume 283, pages 216–218 (1980)Cite this article
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Abstract
Plasmid ColE1, like many other small non-conjugative plasmids, is present in multiple copies (about 15 per chromsome equivalent) in Escherichia coli cells1,2. Because of their high copy number, the replication of such plasmids has been described as ‘relaxed’, even though there is good evidence that it is strictly controlled: ColE1 derivatives have characteristic but different copy numbers2 and ColE1 copy-number mutants have been characterised3–5. No plasmid-specified protein is essential for the replication of ColE1 and related plasmids, as extensive replication can occur in chloramphenicol-treated cells6, in plasmid-free chloramphenicol-treated cells transfected with a hybrid ColE1/phage replicon and in vitro in extracts derived from plasmid-free cells8. Nevertheless, it is possible that a plasmid-specified protein is involved in ColE1 replication control in viable cells. Here we show that deletion of a given non-essential region from ColE1-like plasmids results in a raised copy number. Such plasmids are stably maintained and have their copy number returned to normal when a complementing plasmid is present in the same cell, indicating that a plasmid-specified diffusible gene product regulates the plasmid content of ColE1-containing cells. Deletion of the equivalent region from the cloning vector pBR322 gives a derivative which has a raised copy number and which has also lost its origin for conjugal transfer; unlike pBR322, it cannot be mobilised.
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Authors and Affiliations
- School of Biological Sciences, University of Sussex, Falmer, Brighton, UK
Andrea J. Twigg & David Sherratt
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- Andrea J. Twigg
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Twigg, A., Sherratt, D. Trans-complementable copy-number mutants of plasmid ColE1.Nature 283, 216–218 (1980). https://doi.org/10.1038/283216a0
- Received: 06 August 1979
- Accepted: 12 November 1979
- Issue Date: 10 January 1980
- DOI: https://doi.org/10.1038/283216a0