title: Two replication regions in the pJM1 virulence plasmid of the marine pathogen Vibrio anguillarum.
abstract: Vibrio anguillarum is a fish pathogen that causes vibriosis, a serious hemorrhagic septicemia, in wild and cultured fish. Many serotype O1 strains of this bacterium harbor the 65kb plasmid pJM1 carrying the majority of genes encoding the siderophore anguibactin iron transport system that is one of the most important virulence factors of this bacterium. We previously identified a replication region of the pJM1 plasmid named ori1. In this work we determined that ori1 can replicate in Escherichia coli and that the chromosome-encoded proteins DnaB, DnaC and DnaG are essential for its replication whereas PolI, IHF and DnaA are not required. The copy number of the pJM1 plasmid is 1-2, albeit cloned smaller fragments of the ori1 region replicate with higher copy numbers in V. anguillarum while in E. coli we did not observe an obvious difference of the copy numbers of these constructs which were all high. Furthermore, we were able to delete the ori1 region from the pJM1 plasmid and identified a second replication region in pJM1 that we named ori2. This second replication region is located on ORF25 that is within the trans-acting factor (TAFr) region, and showed that it can only replicate in V. anguillarum.
authors.affiliation: Department of Molecular Microbiology and Immunology, Oregon Health and Science University, Portland, OR 97239, USA. nakah@ohsu.edu
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Naka
authors.forename: Qian
authors.initials: Q
authors.lastname: Chen
authors.forename: Yasutami
authors.initials: Y
authors.lastname: Mitoma
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Nakamura
authors.forename: Daniel
authors.initials: D
authors.lastname: McIntosh-Tolle
authors.forename: Alison E
authors.initials: AE
authors.lastname: Gammie
authors.forename: Marcelo E
authors.initials: ME
authors.lastname: Tolmasky
authors.forename: Jorge H
authors.initials: JH
authors.lastname: Crosa
mesh_terms: D002876:Chromosomes, Bacterial; D056915:DNA Copy Number Variations; D004261:DNA Replication; D004926:Escherichia coli / Q000235:genetics / Q000472:pathogenicity; D023061:Gene Order; D016366:Open Reading Frames; D010957:Plasmids / Q000235:genetics*; D018741:Replication Origin; D014733:Vibrio / Q000235:genetics* / Q000472:pathogenicity*; D014774:Virulence