Characterization and complete genome sequence analysis of novel bacteriophage IME-EFm1 infecting Enterococcus faecium

J Gen Virol. 2014 Nov;95(Pt 11):2565-2575. doi: 10.1099/vir.0.067553-0. Epub 2014 Jul 30.

Abstract

We isolated and characterized a novel virulent bacteriophage, IME-EFm1, specifically infecting multidrug-resistant Enterococcus faecium. IME-EFm1 is morphologically similar to members of the family Siphoviridae. It was found capable of lysing a wide range of our E. faecium collections, including two strains resistant to vancomycin. One-step growth tests revealed the host lysis activity of phage IME-EFm1, with a latent time of 30 min and a large burst size of 116 p.f.u. per cell. These biological characteristics suggested that IME-EFm1 has the potential to be used as a therapeutic agent. The complete genome of IME-EFm1 was 42 597 bp, and was linear, with terminally non-redundant dsDNA and a G+C content of 35.2 mol%. The termini of the phage genome were determined with next-generation sequencing and were further confirmed by nuclease digestion analysis. To our knowledge, this is the first report of a complete genome sequence of a bacteriophage infecting E. faecium. IME-EFm1 exhibited a low similarity to other phages in terms of genome organization and structural protein amino acid sequences. The coding region corresponded to 90.7 % of the genome; 70 putative ORFs were deduced and, of these, 29 could be functionally identified based on their homology to previously characterized proteins. A predicted metallo-β-lactamase gene was detected in the genome sequence. The identification of an antibiotic resistance gene emphasizes the necessity for complete genome sequencing of a phage to ensure it is free of any undesirable genes before use as a therapeutic agent against bacterial pathogens.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteriophages / genetics*
  • Bacteriophages / isolation & purification
  • Bacteriophages / pathogenicity*
  • Base Sequence
  • DNA, Viral / genetics
  • Drug Resistance, Multiple, Bacterial
  • Enterococcus faecium / drug effects
  • Enterococcus faecium / virology*
  • Genome, Viral
  • Host Specificity
  • Molecular Sequence Data
  • Open Reading Frames
  • Phylogeny
  • Siphoviridae / genetics
  • Virulence / genetics

Substances

  • DNA, Viral

Associated data

  • GENBANK/KJ010489