New England Biolabs, Ipswich, Massachusetts.
Genome Biol Evol. 2019 Mar 1;11(3):706-720. doi: 10.1093/gbe/evz025.
Wolbachia, an alpha-proteobacterium closely related to Rickettsia, is a maternally transmitted, intracellular symbiont of arthropods and nematodes. Aedes albopictus mosquitoes are naturally infected with Wolbachia strains wAlbA and wAlbB. Cell line Aa23 established from Ae. albopictus embryos retains only wAlbB and is a key model to study host-endosymbiont interactions. We have assembled the complete circular genome of wAlbB from the Aa23 cell line using long-read PacBio sequencing at 500× median coverage. The assembled circular chromosome is 1.48 megabases in size, an increase of more than 300 kb over the published draft wAlbB genome. The annotation of the genome identified 1,205 protein coding genes, 34 tRNA, 3 rRNA, 1 tmRNA, and 3 other ncRNA loci. The long reads enabled sequencing over complex repeat regions which are difficult to resolve with short-read sequencing. Thirteen percent of the genome comprised insertion sequence elements distributed throughout the genome, some of which cause pseudogenization. Prophage WO genes encoding some essential components of phage particle assembly are missing, while the remainder are found in five prophage regions/WO-like islands or scattered around the genome. Orthology analysis identified a core proteome of 535 orthogroups across all completed Wolbachia genomes. The majority of proteins could be annotated using Pfam and eggNOG analyses, including ankyrins and components of the Type IV secretion system. KEGG analysis revealed the absence of five genes in wAlbB which are present in other Wolbachia. The availability of a complete circular chromosome from wAlbB will enable further biochemical, molecular, and genetic analyses on this strain and related Wolbachia.
沃尔巴克氏体(Wolbachia),一种与立克次氏体密切相关的α-变形菌,是节肢动物和线虫的一种母系传递、细胞内共生体。白纹伊蚊(Aedes albopictus)自然感染沃尔巴克氏体菌株 wAlbA 和 wAlbB。从白纹伊蚊胚胎中建立的细胞系 Aa23 仅保留 wAlbB,是研究宿主-共生体相互作用的关键模型。我们使用长读长 PacBio 测序,以 500×中位数覆盖度,从 Aa23 细胞系中组装了 wAlbB 的完整圆形基因组。组装的圆形染色体大小为 1.48 兆碱基,比已发表的 wAlbB 基因组草案增加了 300 多千碱基。基因组注释鉴定了 1205 个蛋白编码基因、34 个 tRNA、3 个 rRNA、1 个 tmRNA 和 3 个其他非编码 RNA 基因座。长读长测序能够对复杂重复区域进行测序,而这些区域很难通过短读长测序来解析。基因组的 13%由插入序列元件组成,这些元件分布在整个基因组中,其中一些导致了假基因化。编码噬菌体粒子组装一些必需成分的噬菌体 WO 基因缺失,而其余的基因则存在于五个噬菌体区域/ WO 样岛或散布在基因组周围。同源分析确定了所有已完成的沃尔巴克氏体基因组中 535 个直系同源物的核心蛋白质组。大多数蛋白质可以使用 Pfam 和 eggNOG 分析进行注释,包括锚蛋白和 IV 型分泌系统的组成部分。KEGG 分析显示 wAlbB 中缺失了五个基因,这些基因存在于其他沃尔巴克氏体中。wAlbB 完整圆形染色体的可用性将使我们能够对该菌株和相关沃尔巴克氏体进行进一步的生化、分子和遗传分析。