Multimerization of high copy number plasmids causes instability: CoIE1 encodes a determinant essential for plasmid monomerization and stability

Cell. 1984 Apr;36(4):1097-103. doi: 10.1016/0092-8674(84)90060-6.

Abstract

Although the natural multicopy plasmid CoIE1 is maintained stably under most growth conditions, plasmid cloning vectors related to it are relatively unstable, being lost at frequencies of 10(-2)-10(-5) per cell per generation. Evidence suggests that CoIE1 and related plasmids are partitioned randomly at cell division and that plasmid stability is correlated inversely with plasmid multimerization; factors or conditions that reduce multimerization increase stability. Cells containing plasmid multimers segregate plasmid-free cells because the multimers are maintained at lower copy numbers than monomers, as predicted by origin-counting models for copy number control. CoIE1 is stable because it encodes a determinant, cer, that is necessary for recA-, recF-, and recE-independent recombination events that efficiently convert any multimers to monomers. We have localized monomerizing and stability determinants of CoIE1 to within a 0.38 kb region that, when cloned into plasmid vectors, greatly increases their stability.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ampicillin / toxicity
  • Bacteriocin Plasmids*
  • Cloning, Molecular
  • DNA Restriction Enzymes
  • Escherichia coli / genetics*
  • Genetic Vectors
  • Kinetics
  • Penicillin Resistance
  • Plasmids*

Substances

  • Ampicillin
  • DNA Restriction Enzymes