title: Genomic characterization of JG068, a novel virulent podovirus active against Burkholderia cenocepacia.
abstract: BACKGROUND
  As is true for many other antibiotic-resistant Gram-negative pathogens, members of the Burkholderia cepacia complex (BCC) are currently being assessed for their susceptibility to phage therapy as an antimicrobial treatment. The objective of this study was to perform genomic and limited functional characterization of the novel BCC phage JG068 (vB_BceP_JG068).
  
  
  RESULTS
  JG068 is a podovirus that forms large, clear plaques on Burkholderia cenocepacia K56-2. Host range analysis indicates that this phage can infect environmental, clinical, and epidemic isolates of Burkholderia multivorans, B. cenocepacia, Burkholderia stabilis, and Burkholderia dolosa, likely through interaction with the host lipopolysaccharide as a receptor. The JG068 chromosome is 41,604 base pairs (bp) in length and is flanked by 216 bp short direct terminal repeats. Gene expression originates from both host and phage promoters and is in the forward direction for all 49 open reading frames. The genome sequence shows similarity to Ralstonia phage ϕRSB1, Caulobacter phage Cd1, and uncharacterized genetic loci of blood disease bacterium R229 and Burkholderia pseudomallei 1710b. CoreGenesUniqueGenes analysis indicates that JG068 belongs to the Autographivirinae subfamily and ϕKMV-like phages genus. Modules within the genome encode proteins involved in DNA-binding, morphogenesis, and lysis, but none associated with pathogenicity or lysogeny. Similar to the signal-arrest-release (SAR) endolysin of ϕKMV, inducible expression of the JG068 SAR endolysin causes lysis of Escherichia coli that is dependent on the presence of an N-terminal signal sequence. In an in vivo assay using the Galleria mellonella infection model, treatment of B. cenocepacia K56-2-infected larvae with JG068 results in a significant increase in larval survival.
  
  
  CONCLUSIONS
  As JG068 has a broad host range, does not encode virulence factors, is obligately lytic, and has activity against an epidemic B. cenocepacia strain in vivo, this phage is a highly promising candidate for BCC phage therapy development.
authors: Department of Biological Sciences, 6-008 Centennial Centre for Interdisciplinary Science, University of Alberta, Edmonton, AB T6G 2E9 Canada. jon.dennis@ualberta.ca.
authors: Karlene H
authors: KH
authors: Lynch
authors: Ashraf H
authors: AH
authors: Abdu
authors: Max
authors: M
authors: Schobert
authors: Jonathan J
authors: JJ
authors: Dennis
mesh_terms: D001483:Base Sequence; D057508:Burkholderia cenocepacia / Q000821:virology*; D016679:Genome, Viral; D058507:Host Specificity; D058977:Molecular Sequence Annotation; D008969:Molecular Sequence Data; D017902:Podoviridae / Q000235:genetics* / Q000302:isolation & purification / Q000648:ultrastructure; D011401:Promoter Regions, Genetic; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D012722:Sewage / Q000821:virology; D012988:Soil Microbiology; D013728:Terminator Regions, Genetic; D024363:Transcription Initiation Site; D014764:Viral Proteins / Q000235:genetics; D014771:Virion / Q000235:genetics / Q000302:isolation & purification / Q000648:ultrastructure; D014774:Virulence
chemical_list: D012722:Sewage; D014764:Viral Proteins
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