Inactivation of Burkholderia cepacia complex phage KS9 gp41 identifies the phage repressor and generates lytic virions.
The Burkholderia cepacia complex (BCC) is made up of at least 17 species of gram-negative opportunistic bacterial pathogens that cause fatal infections in patients with cystic fibrosis and chronic granulomatous disease. KS9 (vB_BcenS_KS9), one of a number of temperate phages isolated from BCC species, is a prophage of Burkholderia pyrrocinia LMG 21824. Transmission electron micrographs indicate that KS9 belongs to the family Siphoviridae and exhibits the B1 morphotype. The 39,896-bp KS9 genome, comprised of 50 predicted genes, integrates into the 3' end of the LMG 21824 GTP cyclohydrolase II open reading frame. The KS9 genome is most similar to uncharacterized prophage elements in the genome of B. cenocepacia PC184 (vB_BcenZ_ PC184), as well as Burkholderia thailandensis phage phiE125 and Burkholderia pseudomallei phage phi1026b. Using molecular techniques, we have disrupted KS9 gene 41, which exhibits similarity to genes encoding phage repressors, producing a lytic mutant named KS9c. This phage is incapable of stable lysogeny in either LMG 21824 or B. cenocepacia strain K56-2 and rescues a Galleria mellonella infection model from experimental B. cenocepacia K56-2 infections at relatively low multiplicities of infection. These results readily demonstrate that temperate phages can be genetically engineered to lytic form and that these modified phages can be used to treat bacterial infections in vivo.
Lynch
Karlene H
KH
Department of Biological Sciences, University of Alberta, Edmonton, Alberta T6G 2E9, Canada.
Seed
Kimberley D
KD
Stothard
Paul
P
Dennis
Jonathan J
JJ
D001435:Bacteriophages / Q000235:genetics / Q000502:physiology* / Q000648:ultrastructure; D001483:Base Sequence; D016956:Burkholderia cepacia / Q000472:pathogenicity / Q000821:virology*; D017931:DNA Primers; D016679:Genome, Viral; D046529:Microscopy, Electron, Transmission; D009690:Nucleic Acid Conformation; D016366:Open Reading Frames; D012097:Repressor Proteins / Q000502:physiology*; D010948:Viral Plaque Assay; D014771:Virion / Q000502:physiology*; D014774:Virulence; D038301:Virus Inactivation
D017931:DNA Primers; D012097:Repressor Proteins