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1.
Viruses ; 7(4): 1804-22, 2015 Apr 07.
Article in English | MEDLINE | ID: mdl-25853484

ABSTRACT

Klebsiella pneumoniae phages vB_KpnP_SU503 (SU503) and vB_KpnP_SU552A (SU552A) are virulent viruses belonging to the Autographivirinae subfamily of Podoviridae that infect and kill multi-resistant K. pneumoniae isolates. Phages SU503 and SU552A show high pairwise nucleotide identity to Klebsiella phages KP34 (NC_013649), F19 (NC_023567) and NTUH-K2044-K1-1 (NC_025418). Bioinformatic analysis of these phage genomes show high conservation of gene arrangement and gene content, conserved catalytically active residues of their RNA polymerase, a common and specific lysis cassette, and form a joint cluster in phylogenetic analysis of their conserved genes. Also, we have performed biological characterization of the burst size, latent period, host specificity (together with KP34 and NTUH-K2044-K1-1), morphology, and structural genes as well as sensitivity testing to various conditions. Based on the analyses of these phages, the creation of a new phage genus is suggested within the Autographivirinae, called "Kp34likevirus" after their type phage, KP34. This genus should encompass the recently genome sequenced Klebsiella phages KP34, SU503, SU552A, F19 and NTUH-K2044-K1-1.


Subject(s)
Bacteriophages/classification , Klebsiella/virology , Podoviridae/classification , Bacteriophages/genetics , Bacteriophages/growth & development , Bacteriophages/ultrastructure , Computational Biology , Electrophoresis, Polyacrylamide Gel , Gene Order , Genes, Viral , Host Specificity , Microscopy, Electron, Transmission , Podoviridae/genetics , Podoviridae/growth & development , Podoviridae/ultrastructure , Synteny , Viral Structural Proteins/analysis , Virion/chemistry , Virion/ultrastructure
2.
PLoS One ; 9(12): e116294, 2014.
Article in English | MEDLINE | ID: mdl-25551446

ABSTRACT

A recently isolated phage, vB_EcoP_SU10 (SU10), with the unusual elongated C3 morphotype, can infect a wide range of Escherichia coli strains. We have sequenced the genome of this phage and characterized it further by mass spectrometry based proteomics, transmission electron microscopy (TEM), scanning electron microscopy (SEM), and ultra-thin section electron microscopy. The genome size is 77,327 base pairs and its genes, and genome architecture, show high similarity to the phiEco32 phage genes and genome. The TEM images reveal that SU10 have a quite long tail for being a Podoviridae phage, and that the tail also changes conformation upon infection. The ultra-thin section electron microscopy images of phages at the stage of replication within the host cell show that the phages form a honeycomb-like structure under packaging of genomes and assembly of mature capsids. This implies a tight link between the replication and cutting of the concatemeric genome, genome packaging, and capsid assembly. We have also performed a phylogenetic analysis of the structural genes common between Podoviridae phages of the C1 and C3 morphotypes. The result shows that the structural genes have coevolved, and that they form two distinct groups linked to their morphotypes. The structural genes of C1 and C3 phages appear to have diverged around 280 million years ago applying a molecular clock calibrated according to the presumed split between the Escherichia - Salmonella genera.


Subject(s)
Genome, Viral , Phylogeny , Podoviridae/genetics , Proteomics , Chromosome Mapping , DNA, Viral/chemistry , Genomics , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , Molecular Sequence Data , Podoviridae/ultrastructure , Sequence Analysis, DNA , Viral Proteins/chemistry , Viral Proteins/genetics
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