Plasmid incompatibility: more compatible than previously thought?
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.
Velappan
Nileena
N
Biosciences Division, Los Alamos National Laboratory, Los Alamos, NM 87545, USA.
Sblattero
Daniele
D
Chasteen
Leslie
L
Pavlik
Peter
P
Bradbury
Andrew R M
AR
D000668:Ampicillin Resistance / Q000235:genetics; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000821:virology; D007613:Kanamycin Resistance / Q000235:genetics; D010957:Plasmids / Q000235:genetics*; D018741:Replication Origin; D013753:Tetracycline Resistance / Q000235:genetics