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鉴定耶尔森噬菌体 φR1-37 的基因组、蛋白质组和结构。

Characterization of the genome, proteome, and structure of yersiniophage ϕR1-37.

机构信息

Department of Bacteriology and Immunology, Infection Biology Research Programme, Haartman Institute, University of Helsinki, Helsinki, Finland.

出版信息

J Virol. 2012 Dec;86(23):12625-42. doi: 10.1128/JVI.01783-12. Epub 2012 Sep 12.

Abstract

The bacteriophage vB_YecM-ϕR1-37 (ϕR1-37) is a lytic yersiniophage that can propagate naturally in different Yersinia species carrying the correct lipopolysaccharide receptor. This large-tailed phage has deoxyuridine (dU) instead of thymidine in its DNA. In this study, we determined the genomic sequence of phage ϕR1-37, mapped parts of the phage transcriptome, characterized the phage particle proteome, and characterized the virion structure by cryo-electron microscopy and image reconstruction. The 262,391-bp genome of ϕR1-37 is one of the largest sequenced phage genomes, and it contains 367 putative open reading frames (ORFs) and 5 tRNA genes. Mass-spectrometric analysis identified 69 phage particle structural proteins with the genes scattered throughout the genome. A total of 269 of the ORFs (73%) lack homologues in sequence databases. Based on terminator and promoter sequences identified from the intergenic regions, the phage genome was predicted to consist of 40 to 60 transcriptional units. Image reconstruction revealed that the ϕR1-37 capsid consists of hexameric capsomers arranged on a T=27 lattice similar to the bacteriophage ϕKZ. The tail of ϕR1-37 has a contractile sheath. We conclude that phage ϕR1-37 is a representative of a novel phage type that carries the dU-containing genome in a ϕKZ-like head.

摘要

噬菌体 vB_YecM-ϕR1-37(ϕR1-37)是一种裂解性耶尔森噬菌体,能够在携带正确脂多糖受体的不同耶尔森氏菌中自然繁殖。这种长尾噬菌体的 DNA 中含有脱氧尿嘧啶(dU)而不是胸腺嘧啶。在本研究中,我们确定了噬菌体 ϕR1-37 的基因组序列,绘制了部分噬菌体转录组图谱,表征了噬菌体粒子的蛋白质组,并通过冷冻电镜和图像重建对病毒粒子结构进行了表征。噬菌体 ϕR1-37 的 262391bp 基因组是已测序的最大噬菌体基因组之一,其中包含 367 个推定的开放阅读框(ORF)和 5 个 tRNA 基因。质谱分析鉴定了 69 种噬菌体粒子结构蛋白,这些基因散布在整个基因组中。ORF 总数(73%)的 269 个(73%)在序列数据库中没有同源物。根据在基因间区识别的终止子和启动子序列,预测噬菌体基因组由 40 到 60 个转录单元组成。图像重建表明,噬菌体 ϕR1-37 的衣壳由六聚体衣壳组成,排列在 T=27 晶格上,类似于噬菌体 ϕKZ。噬菌体 ϕR1-37 的尾巴有一个可收缩的鞘。我们得出结论,噬菌体 ϕR1-37 是一种新型噬菌体的代表,它在类似于 ϕKZ 的头部携带含有 dU 的基因组。

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