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感染猪丹毒丝菌的温和噬菌体SE-I的基因组和蛋白质组特征分析

Genomic and proteomic characterization of SE-I, a temperate bacteriophage infecting Erysipelothrix rhusiopathiae.

作者信息

Yuan Wentao, Zhang Yaning, Wang Guangcao, Bai Juan, Wang Xianwei, Li Yufeng, Jiang Ping

机构信息

College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, China.

出版信息

Arch Virol. 2016 Nov;161(11):3137-50. doi: 10.1007/s00705-016-3018-1. Epub 2016 Aug 19.

Abstract

A bacteriophage infecting pathogenic Erysipelothrix rhusiopathiae was isolated from a swine farm experiencing an outbreak of acute swine erysipelas; we designated this phage SE-I. SE-I has an icosahedral head, a long tail and a double-stranded DNA genome. The 34,997-bp genome has a GC content of 34 % and contains 43 open reading frames (ORFs) encoding packaging, structural, lysin-holin, and hypothetical proteins. Components of purified SE-I were separated using SDS-PAGE and analyzed using liquid chromatography-mass spectrometry. Nine proteins were identified, encoded by ORF9, ORF15, ORF23, ORF30, ORF31, ORF33, ORF39, ORF40 and ORF 42. A phylogenetic tree constructed based on the sequence of the large terminase subunit revealed that SE-I is closely related to Staphylococcus phages P954 and phi3396. The CHAP-domain-containing protein encoded by ORF25 was expressed in E. coli and which was able to inactivate host bacteria. SE-I was able to infect 7 of 13 E. rhusiopathiae strains, but was unable to infect Salmonella, Streptococcus suis, and Staphylococcus aureus. This is the first report of the isolation, characterization, and genomic and proteomic analysis of a temperate phage infecting E. rhusiopathiae, and it might lead to the development of new anti- E. rhusiopathiae agents.

摘要

从一个爆发急性猪丹毒的养猪场中分离出一种感染致病性猪丹毒丝菌的噬菌体;我们将这种噬菌体命名为SE-I。SE-I具有二十面体头部、长尾部和双链DNA基因组。34997碱基对的基因组GC含量为34%,包含43个开放阅读框(ORF),编码包装、结构、溶菌酶-穿孔素和假定蛋白。使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分离纯化的SE-I的组分,并使用液相色谱-质谱联用仪进行分析。鉴定出9种蛋白质,由ORF9、ORF15、ORF23、ORF30、ORF31、ORF33、ORF39、ORF40和ORF42编码。基于大末端酶亚基序列构建的系统发育树表明,SE-I与葡萄球菌噬菌体P954和phi3396密切相关。由ORF25编码的含CHAP结构域的蛋白质在大肠杆菌中表达,并且能够使宿主细菌失活。SE-I能够感染13株猪丹毒丝菌中的7株,但不能感染沙门氏菌、猪链球菌和金黄色葡萄球菌。这是关于感染猪丹毒丝菌的温和噬菌体的分离、表征以及基因组和蛋白质组分析的首次报道,它可能会促成新型抗猪丹毒丝菌药物的开发。

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