Detection of Deazapurine modifications in phage DNA using nanopore sequencing

Research output: Contribution to conferencePosterResearch

  • Witold Kot
  • ,
  • Nikoline S. Olsen
  • Tue Kjærgaard Nielsen
  • ,
  • Geoffrey Hutinet, Genetics Institute, University of Florida, United States
  • Valérie de Crécy-Lagard, Genetics Institute, University of Florida, United States
  • Peter Dedon, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, United States
  • Alexander Byth Carstens
  • ,
  • Sylvain Moineau, Faculté des Sciences et de Génie, Université Laval
  • ,
  • Lars Hestbjerg Hansen
Bacteria, continually exposed to bacteriophages, have developed several defense mechanisms e.g. restriction-modification or CRISPR-Cas systems. On the other hand, bacteriophages have developed several strategies to evade these echanisms.
Escherichia coli virulent siphophage CAjan, belonging to Seuratvirus genus and the closely related genus nonagvirus, contains a homolog of the tRNA-deazapurine modification synthesis pathway. The evidence is piling up, that CAjan and other similar bacteriophages use these deazapurine modifications in their DNA to evade bacterial restriction-modification systems. In order to investigate this novel DNA modification pathway in detail we have used several methods including; direct phage plaque sequencing, CRISPR-Cas editing of phage genome and nanopore sequencing of viral DNA. Through generation of specific mutants within the deazapurine modification syntheis pathway followed by nanopore seqeuncing, we were able to 1) obtain a restriction sensitive phenotype in the CAjan bacteriophage and 2) detect the modified bases using nanopore seqeuncing, thus providing new insights on use of alternative bases by bacteriophages.
Original languageEnglish
Publication year8 Jul 2018
Number of pages1
Publication statusPublished - 8 Jul 2018
EventViruses of Microbes: 5th Viruses of Microbes meeting - University of Wroclaw, Wroclaw, Poland
Duration: 9 Jul 201813 Jul 2018
http://meetings.embo.org/event/18-virus-microbe

Conference

ConferenceViruses of Microbes
LocationUniversity of Wroclaw
CountryPoland
CityWroclaw
Period09/07/201813/07/2018
Internet address

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