Abstract
The bacteriocinogenic plasmid Clo DF13, originally isolated from Enterobacter cloacae1, is stably maintained in Escherichia coli2 to the extent of about 10 copies per cell3. Its replication4 resembles that of many other small, multicopy plasmids5–8; plasmid-encoded protein is not required9–12 but plasmid-specific genetic information is involved in regulation of replication as both conditional13 and nonconditional3 copy-number mutants of Clo DF13, and transcomplementable14,15 copy-number mutants of plasmid Col E1 have been described. The sequences essential for replication of Col El (refs 16, 17) and Clo DF13 (refs 18, 19) have been identified within a region surrounding the replication origin. Initiation of Col E1 replication is preceded by transcription of the origin region, providing the RNA primer at the origin20. However, transcription in the opposite direction results in a small transcript of ∼100 nucleotides (RNA-100) for both Col E1 (refs 21, 22) and Clo DF13 (ref. 23). Data suggest that Col E1 RNA-100 acts as a negative control element for the initiation of replication20,24. We show here that single base transitions in the RNA-100 cistron of Clo DF13 can result in a nonconditional increase in plasmid copy-number. Also, sequence analysis has revealed that a specific base transition in a DNA region, apparently involved in both termination and initiation of transcription towards the replication origin, results in a thermosensitive plasmid copy-number.
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Stuitje, A., Spelt, C., Veltkamp, E. et al. Identification of mutations affecting replication control of plasmid Clo DF13. Nature 290, 264–267 (1981). https://doi.org/10.1038/290264a0
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DOI: https://doi.org/10.1038/290264a0
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