Przondovirus属中的噬菌体(噬菌体)是属于Studiervirinae亚科的T7样podovirus,在自拟病毒科中,并且具有高度保守的基因组组织。这些噬菌体的基因组大小从37到42kb,编码50-60个基因,其特征在于存在线性染色体侧翼的直接末端重复(DTR)。这些DTR通常在短只读和混合组装期间被删除。此外,长只读组件通常会出现测序和/或组装错误,需要额外的管理。这里,我们介绍了十种针对克雷伯菌属的新型przondovirus的分离和表征。我们描述了HYPPA,混合和聚波兰噬菌体组装工作流程,它利用长读取组件与短读取测序相结合来解析噬菌体DTR并纠正错误,否定了费力的引物行走和Sanger测序验证的需要。我们的组装工作流程利用牛津纳米孔技术进行长读取测序,使其成为目前更相关的长读数测序技术,和IlluminaDNA准备进行短读测序,代表全球最常用的技术。我们的数据证明了在发表之前仔细管理噬菌体组装的重要性,在将它们用于比较基因组学之前。
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.