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Genome sequence of the phage clP1, which infects the beer spoilage bacterium Pediococcus damnosus
Authors:Kelly David  O'Sullivan Orla  Mills Susan  McAuliffe Olivia  Ross R Paul  Neve Horst  Coffey Aidan
Affiliation:Department of Biological Sciences, Cork Institute of Technology, Co. Cork, Ireland.
Abstract:
Pediococcus damnosus (P. damnosus) bacteriophage (phage) clP1 is a novel virulent phage isolated from a municipal sewage sample collected in Southern Ireland. This phage infects the beer spoilage strain P. damnosus P82 which was isolated from German breweries. Sequencing of the phage has revealed a linear double stranded DNA genome of 38,013 base pairs (bp) with an overall GC content of 47.6%. Fifty seven open reading frames (ORFs) were identified of which 30 showed homology to previously sequenced proteins, and as a consequence 20 of these were assigned predicted functions. The majority of genes displayed homology with genes from the Lactobacillus plantarum phage phiJL-1. All genes were located on the same coding strand and in the same orientation. Morphological characterisation placed phage clP1 as a member of the Siphoviridae family with an isometric head (59 nm diameter) and non-contractile tail (length 175 nm; diameter 10nm. Interestingly, the phage clP1 genome was found to share very limited identity with other phage genome sequences in the database, and was hence considered unique. This was highlighted by the genome organisation which differed slightly to the consensus pattern of genomic organisation usually found in Siphoviridae phages. With the genetic machinery present for a lytic lifecycle and the absence of potential endotoxin factors, this phage may have applications in the biocontrol of beer spoilage bacteria. To our knowledge, this study represents the first reported P. damnosus phage genome sequence.
Keywords:DNA, Deoxyribonucleic acid   RNA, ribonucleic acid   bp, base pair   GC, guanine–cytosine   ORF, Open Reading Frame   nm, nanometer   LAB, lactic acid bacteria   MRS, de Man, Rogosa and Sharpe   PCR, polymerase chain reaction   rRNA, ribosomal RNA   μl, microliters   mM, milliMolar   p-mol, picomoles   u, units   min, minutes   s, seconds   °C, Celsius degrees   BLAST, Basic Local Alignment Search Tool   g, grams   ml, Milliliters   CaCl2, Calcium chloride   rpm, Revolutions per minute   μm, micrometers   pfu, plaque-forming unit(s)   w/v, weight/volume   kV, kilovolts   M, Molar   Tris, tris(hydroxymethyl)aminomethane   HCl, hydrochloric acid   MgCl2, magnesium chloride   SDS, sodium dodecyl sulfate   β, beta   V, Volts   RNAse, ribonuclease   EDTA, ethylenediaminetetraacetic acid   ng, nanograms   tRNA, transfer ribonucleic acid   mRNA, messenger ribonucleic acid   tmRNA, transfer-messenger RNA   ACT, Artemis Comparison Tool   PSI, Position Specific Iterative   nr, non-redundant   mm, millimeters   kDa, kilodaltons   PAGE, polyacrylamide gel electrophoresis   Φ, Phi   MW, molecular weight
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