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Elek, Claire K. A., Brown, Teagan L., Viet, Thanh Le, Evans, Rhiannon, Baker, David J., Telatin, Andrea, Tiwari, Sumeet K., Al-Khanaq, Haider, Thilliez, Gaëtan, Kingsley, Robert A. 
ORCID: https://orcid.org/0000-0002-0194-6485, Hall, Lindsay J. ORCID: https://orcid.org/0000-0001-8938-5709, Webber, Mark A. and Adriaenssens, Evelien M.
(2023)
A hybrid and poly-polish workflow for the complete and accurate assembly of phage genomes: a case study of ten przondoviruses.
Microbial Genomics, 9 (7).
ISSN 2057-5858
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Abstract
Bacteriophages (phages) within the genus Przondovirus are T7-like podoviruses belonging to the subfamily Studiervirinae, within the family Autographiviridae, and have a highly conserved genome organisation. The genomes of these phages range from 37 to 42 kb in size, encode 50–60 genes and are characterised by the presence of direct terminal repeats (DTRs) flanking the linear chromosome. These DTRs are often deleted during short-read-only and hybrid assemblies. Moreover, long-read-only assemblies are often littered with sequencing and/or assembly errors and require additional curation. Here, we present the isolation and characterisation of ten novel przondoviruses targeting Klebsiella spp. We describe HYPPA, a HYbrid and Poly-polish Phage Assembly workflow, which utilises long-read assemblies in combination with short-read sequencing to resolve phage DTRs and correcting errors, negating the need for laborious primer walking and Sanger sequencing validation. Our assembly workflow utilised Oxford Nanopore Technologies for long-read sequencing for its accessibility, making it the more relevant long-read sequencing technology at this time, and Illumina DNA Prep for short-read sequencing, representing the most commonly used technologies globally. Our data demonstrate the importance of careful curation of phage assemblies before publication, and prior to using them for comparative genomics.
| Item Type: | Article |
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| Additional Information: | Funding Information: CKAE is supported by the Medical Research Council (MRC) and JAFRAL as part of the Doctoral Antimicrobial Research Training (DART) MRC iCASE Programme, grant no. MR/R015937/1. TLB, AT, SKT and EMA gratefully acknowledge funding by the Biotechnology and Biological Sciences Research Council (BBSRC); this research was funded by the BBSRC Institute Strategic Programme Gut Microbes and Health BB/R012490/1 and its constituent projects BBS/E/F/000PR10353 and BBS/E/F/000PR10356. TLV, DJB and RE were supported by the Quadram Institute Bioscience BBSRC funded Core Capability Grant (project number BB/CCG1860/1). GT, HAK, RAK and MAW are supported by the BBSRC Institute Strategic Programme Microbes in the Food Chain BB/R012504/1 and its constituent projects BBS/E/F/000PR10348 and BBS/E/F/000PR10349. LJH is supported by Wellcome Trust Investigator Awards 100974 /C/13/Z and 220876/Z/20/Z; by the BBSRC Institute Strategic Programme Gut Microbes and Health BB/R012490/1, and its constituent projects BBS/E/F/000PR10353 and BBS/E/F/000PR10356. |
| Uncontrolled Keywords: | klebsiella,przondovirus,assembly,bacteriophage,phage,sequencing,epidemiology,microbiology,molecular biology,genetics ,/dk/atira/pure/subjectarea/asjc/2700/2713 |
| Faculty \ School: | Faculty of Medicine and Health Sciences > Norwich Medical School Faculty of Science > School of Biological Sciences |
| Related URLs: | |
| Depositing User: | LivePure Connector |
| Date Deposited: | 01 Aug 2023 12:31 |
| Last Modified: | 04 Nov 2023 03:37 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/92752 |
| DOI: | 10.1099/mgen.0.001065 |
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