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PEDS Advance Access originally published online on March 1, 2007
Protein Engineering Design and Selection 2007 20(7):309-313; doi:10.1093/protein/gzm005
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© The Author 2007. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Methods

Plasmid incompatibility: more compatible than previously thought?

Nileena Velappan1,3, Daniele Sblattero2,3, Leslie Chasteen1, Peter Pavlik1 and Andrew R.M. Bradbury1,4

1Biosciences Division, Los Alamos National Laboratory, Los Alamos, USA 2Department of Medical Sciences, University of Eastern Piedmont, Novara, Italy 3These authors contributed equally to this work

4 To whom correspondence should be addressed. E-mail: amb{at}lanl.gov

It is generally accepted that plasmids containing the same origin of replication are incompatible. We have re-examined this concept in terms of the plasmid copy number, by introducing plasmids containing the same origin of replication and different antibiotic resistance genes into bacteria. By selecting for resistance to only one antibiotic, we were able to examine the persistence of plasmids carrying resistances to other antibiotics. We find that plasmids are not rapidly lost, but are able to persist in bacteria for multiple overnight growth cycles, with some dependence upon the nature of the antibiotic selected for. By carrying out the experiments with different origins of replication, we have been able to show that higher copy number leads to longer persistence, but even with low copy plasmids, persistence occurs to a significant degree. This observation holds significance for the field of protein engineering, as the presence of two or more plasmids within bacteria weakens, and confuses, the connection between screened phenotype and genotype, with the potential to wrongly assign specific phenotypes to incorrect genotypes.

Keywords: Comments, corrections, or additions to this paper are welcome on the peds blog, PEDS methods (www.oxfordjournals.org/PEDSmethods).

Received December 20, 2006; accepted January 9, 2007.


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