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Sequence, genome organization, annotation and proteomics of the thermophilic, 47.7-kb Geobacillus stearothermophilus bacteriophage TP-84 and its classification in the new Tp84virus genus.
Skowron, Piotr M; Kropinski, Andrew M; Zebrowska, Joanna; Janus, Lukasz; Szemiako, Kasjan; Czajkowska, Edyta; Maciejewska, Natalia; Skowron, Malgorzata; Los, Joanna; Los, Marcin; Zylicz-Stachula, Agnieszka.
Afiliación
  • Skowron PM; Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, Gdansk, Poland.
  • Kropinski AM; BioVentures Institute Ltd., Poznan, Poland.
  • Zebrowska J; Departments of Food Science, Molecular and Cellular Biology; and Pathobiology University of Guelph, Guelph, ON, Canada.
  • Janus L; Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, Gdansk, Poland.
  • Szemiako K; Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, Gdansk, Poland.
  • Czajkowska E; BioVentures Institute Ltd., Poznan, Poland.
  • Maciejewska N; Department of Molecular Biotechnology and Microbiology, Faculty of Chemistry, Gdansk University of Technology, Gdansk, Poland.
  • Skowron M; Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, Gdansk, Poland.
  • Los J; Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, Gdansk, Poland.
  • Los M; BioVentures Institute Ltd., Poznan, Poland.
  • Zylicz-Stachula A; BioVentures Institute Ltd., Poznan, Poland.
PLoS One ; 13(4): e0195449, 2018.
Article en En | MEDLINE | ID: mdl-29624616
Bacteriophage TP-84 is a well-characterized bacteriophage of historical interest. It is a member of the Siphoviridae, and infects a number of thermophilic Geobacillus (Bacillus) stearothermophilus strains. Its' 47.7-kbp double-stranded DNA genome revealed the presence of 81 coding sequences (CDSs) coding for polypeptides of 4 kDa or larger. Interestingly, all CDSs are oriented in the same direction, pointing to a dominant transcription direction of one DNA strand. Based on a homology search, a hypothetical function could be assigned to 31 CDSs. No RNA or DNA polymerase-coding genes were found on the bacteriophage genome indicating that TP-84 relies on the host's transcriptional and replication enzymes. The TP84 genome is tightly packed with CDSs, typically spaced by several-to-tens of bp or often overlapping. The genome contains five putative promoter-like sequences showing similarity to the host promoter consensus sequence and allowing for a 2-bp mismatch. In addition, ten putative rho-independent terminators were detected. Because the genome sequence shows essentially no similarity to any previously characterised bacteriophage, TP-84 should be considered a new species in an undefined genus within the Siphoviridae family. Thus a taxonomic proposal of a new Tp84virus genus has been accepted by the International Committee on Taxonomy of Viruses. The bioinformatics genome analysis was verified by confirmation of 33 TP-84 proteins, which included: a) cloning of a selected CDS in Escherichia coli, coding for a DNA single-stranded binding protein (SSB; gene TP84_63), b) purification and functional assays of the recombinant TP-84 SSB, which has been shown to improve PCR reactions, c) mass spectrometric (MS) analysis of TP-84 bacteriophage capsid proteins, d) purification of TP-84 endolysin activity, e) MS analysis of the host cells from infection time course.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Geobacillus stearothermophilus / Genoma Viral / Siphoviridae Idioma: En Revista: PLoS One Asunto de la revista: CIENCIA / MEDICINA Año: 2018 Tipo del documento: Article País de afiliación: Polonia Pais de publicación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Geobacillus stearothermophilus / Genoma Viral / Siphoviridae Idioma: En Revista: PLoS One Asunto de la revista: CIENCIA / MEDICINA Año: 2018 Tipo del documento: Article País de afiliación: Polonia Pais de publicación: Estados Unidos