Escherichia coli ColV plasmid pRK100: genetic organization, stability and conjugal transfer

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Abstract
UropathogenicEscherichia colistrains express chromosomal and plasmid-encoded virulence-associated factors such as specific adhesins, toxins and iron-uptake systems. A ColV plasmid (pRK100) of a uropathogenic strain and its host KS533 were studied. The host strain encodes the K1 capsule, and P and S fimbriae, but neither haemolysin nor the cytotoxic-necrotic factor CNF1, indicating that this strain does not harbour a larger pathogenicity island. A restriction map of pRK100 was constructed on the basis of hybridization experiments and nucleotide sequencing. pRK100 harbours ColV, the conserved replication region RepFIB, the aerobactin-uptake system, a RepFIC replicon and additionally Colla as well as transposon Tn5431. The location of the RepFIC replicon was similar to that in plasmid F. ColV plasmids and F thus share a region spanning more than half the length of plasmid F. Even though their replication and transfer regions are homologous, ColV plasmids are found only inE. colistrains. Among the four other species tested, conjugal transfer of pRK100 was demonstrated, with low frequency, only toKlebsiella pneumoniae, suggesting that a natural barrier effectively bars transfer.In vitrostability of the plasmid with integration into the chromosome to ensure maintenance in the presence of an incompatible plasmid was demonstrated.