Maintenance of bacterial plasmids: Comparison of theoretical calculations and experiments with plasmid R1 in Escherichia coli

Abstract

Plasmid R1 was tested for stable maintenance in Escherichia coli K12. Populations carrying a transfer-negative derivative of plasmid R1drd-19 were grown exponentially for 72 generations in LB medium. Out of nearly 5000 cells none had lost the plasmid; hence the rate of loss of the plasmid is less than 3 X 10(-6) per cell and cell generation. Other experiments showed that the loss rate was less than or equal to 10(-7) per cell and cell generation. Plasmid R1 replication is controlled so that, on average, n copies are synthesized per cell and cell generation irrespective of the copy number (as long as it is at least one). A theoretical analysis was performed, based on the assumption that there is a Poissonian spread around n in each class of cells (each class defined by the number of R1 copies at birth). Two models for partitioning were analyzed, Equi-partitioning and Pair Site Partitioning; in the latter, each daughter is guaranteed one plasmid copy, whereas the rest of the copies are assumed to be randomly distributed. The copy number distribution was found to be fairly narrow in both cases, with at least 90% of the population in the range n/2-3n/2 for baby cells and in the range n-3n for cells just before cell division. Plasmid-free cells were estimated to appear at a frequency which for the copy number of R1 (n = 3-4) was 1.5 X 10(-3) - 8 X 10(-5) for equipartitioning and 6 X 10(-3) - 6 X 10(-4) for pair site partitioning, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)