Characterization and genomic analysis of phage asccphi28, a phage of the family Podoviridae infecting Lactococcus lactis.
Bacteriophage asccphi28 infects dairy fermentation strains of Lactococcus lactis. This report describes characterization of asccphi28 and its full genome sequence. Phage asccphi28 has a prolate head, whiskers, and a short tail (C2 morphotype). This morphology and DNA hybridization to L. lactis phage P369 DNA showed that asccphi28 belongs to the P034 phage species, a group rarely encountered in the dairy industry. The burst size of asccphi28 was found to be 121 +/- 18 PFU per infected bacterial cell after a latent period of 44 min. The linear genome (18,762 bp) contains 28 possible open reading frames (ORFs) comprising 90% of the total genome. The ORFs are arranged bidirectionally in recognizable functional modules. The genome contains 577 bp inverted terminal repeats (ITRs) and putatively eight promoters and four terminators. The presence of ITRs, a phage-encoded DNA polymerase, and a terminal protein that binds to the DNA, along with BLAST and morphology data, show that asccphi28 more closely resembles streptococcal phage Cp-1 and the phi29-like phages that infect Bacillus subtilis than it resembles common lactococcal phages. The sequence of this phage is the first published sequence of a P034 species phage genome.
Kotsonis
Steven E
SE
Department of Biochemistry and Molecular Biology, The University of Melbourne, Melbourne, Victoria 3010, Australia.
Powell
Ian B
IB
Pillidge
Christopher J
CJ
Limsowtin
Gaëtan K Y
GK
Hillier
Alan J
AJ
Davidson
Barrie E
BE
D017103:Bacillus Phages / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics; D004259:DNA-Directed DNA Polymerase / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D023061:Gene Order; D005814:Genes, Viral; D016679:Genome, Viral; D013294:Lactococcus lactis / Q000821:virology*; D046529:Microscopy, Electron, Transmission; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D017902:Podoviridae / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification / Q000648:ultrastructure; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D017108:Streptococcus Phages / Q000235:genetics; D026801:Synteny; D020079:Terminal Repeat Sequences; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000235:genetics; D014771:Virion / Q000648:ultrastructure
D004279:DNA, Viral; D004268:DNA-Binding Proteins; D014764:Viral Proteins; D004259:DNA-Directed DNA Polymerase