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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
3
pubmed:dateCreated
2001-12-13
pubmed:databankReference
pubmed:abstractText
We report here the molecular characterization of pFNL10, a 3990-bp cryptic plasmid of Francisella novicida-like F6168. The plasmid was maintained in F. novicida Utah 112 and F. tularensis LVS strains. We sequenced the entire plasmid and found six open reading frames (ORFs)-ORF1, ORF2, ORF3, ORF4, ORF5, and ORFm. ORF3, ORF4, ORF5, and ORFm are located on the same strand, and we designated it the plus strand. ORF1 and ORF2 are on the complementary strand. The ORFs appear to be arranged in two operons, one comprising ORF5 and ORF4 and the other ORF1 and ORF2. There exist two distinct promoters similar to the Escherichia coli sigma(70) promoter, one 5' to ORF1-ORF2 operon and the other 5' to ORF5-ORF4 operon. We found that in both promoters the transcriptional start is an adenosine. ORF3 is positioned in tandem with ORF5-ORF4, but has its own transcriptional start, a thymidine. However, sequence analysis revealed no recognizable promoter in physical proximity to ORF3. Sequence analysis revealed transcriptional terminators immediately downstream of the two operons. Experimental results showed that the ORF1-ORF2 terminator is authentic. But we could not definitively confirm the ORF5-ORF4 terminator. Two sets of direct repeats, one 31 and the other 13 bp, characteristic of ori are positioned between the two promoters. ORF1 encodes a protein that bears homology to the replication initiation protein RepA of various bacteria, and disruption of this ORF indeed blocked pFNL10 replication. In contrast, ORF2 disruption caused formation of plasmid multimers, suggesting aberrant replication. Our analysis also suggests that pFNL10 replicates by the theta mode. The ORF5-ORF4 operon resembles the phd-doc operon of Escherichia coli bacteriophage P1, but the significance of this similarity is unclear.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Nov
pubmed:issn
0147-619X
pubmed:author
pubmed:copyrightInfo
Copyright 2001 Elsevier Science.
pubmed:issnType
Print
pubmed:volume
46
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
210-22
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:11735370-Amino Acid Sequence, pubmed-meshheading:11735370-Animals, pubmed-meshheading:11735370-Bacterial Proteins, pubmed-meshheading:11735370-Chloramphenicol O-Acetyltransferase, pubmed-meshheading:11735370-DNA, Bacterial, pubmed-meshheading:11735370-DNA Replication, pubmed-meshheading:11735370-Escherichia coli, pubmed-meshheading:11735370-Francisella, pubmed-meshheading:11735370-Francisella tularensis, pubmed-meshheading:11735370-Genes, Bacterial, pubmed-meshheading:11735370-Genes, Reporter, pubmed-meshheading:11735370-Mice, pubmed-meshheading:11735370-Molecular Sequence Data, pubmed-meshheading:11735370-Open Reading Frames, pubmed-meshheading:11735370-Plasmids, pubmed-meshheading:11735370-Species Specificity, pubmed-meshheading:11735370-Terminator Regions, Genetic, pubmed-meshheading:11735370-Transformation, Bacterial, pubmed-meshheading:11735370-Tularemia, pubmed-meshheading:11735370-Virulence
pubmed:year
2001
pubmed:articleTitle
Genetic organization of the Francisella plasmid pFNL10.
pubmed:affiliation
State Research Center for Applied Microbiology, Obolensk Moscow Region, 142279, Russia.
pubmed:publicationType
Journal Article, Comparative Study, Research Support, Non-U.S. Gov't