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苏牧 Mu 样噬菌体的基因组和蛋白质组特征分析,该噬菌体感染副猪嗜血杆菌。

Genomic and proteomic characterization of SuMu, a Mu-like bacteriophage infecting Haemophilus parasuis.

机构信息

U.S. Department of Agriculture, Ruminant Diseases and Immunology, National Animal Disease Center, Agricultural Research Service, Ames, IA 50010, USA.

出版信息

BMC Genomics. 2012 Jul 23;13:331. doi: 10.1186/1471-2164-13-331.

Abstract

BACKGROUND

Haemophilus parasuis, the causative agent of Glässer's disease, is prevalent in swine herds and clinical signs associated with this disease are meningitis, polyserositis, polyarthritis, and bacterial pneumonia. Six to eight week old pigs in segregated early weaning herds are particularly susceptible to the disease. Insufficient colostral antibody at weaning or the mixing of pigs with heterologous virulent H. parasuis strains from other farm sources in the nursery or grower-finisher stage are considered to be factors for the outbreak of Glässer's disease. Previously, a Mu-like bacteriophage portal gene was detected in a virulent swine isolate of H. parasuis by nested polymerase chain reaction. Mu-like bacteriophages are related phyologenetically to enterobacteriophage Mu and are thought to carry virulence genes or to induce host expression of virulence genes. This study characterizes the Mu-like bacteriophage, named SuMu, isolated from a virulent H. parasuis isolate.

RESULTS

Characterization was done by genomic comparison to enterobacteriophage Mu and proteomic identification of various homologs by mass spectrometry. This is the first report of isolation and characterization of this bacteriophage from the Myoviridae family, a double-stranded DNA bacteriophage with a contractile tail, from a virulent field isolate of H. parasuis. The genome size of bacteriophage SuMu was 37,151 bp. DNA sequencing revealed fifty five open reading frames, including twenty five homologs to Mu-like bacteriophage proteins: Nlp, phage transposase-C-terminal, COG2842, Gam-like protein, gp16, Mor, peptidoglycan recognition protein, gp29, gp30, gpG, gp32, gp34, gp36, gp37, gpL, phage tail tube protein, DNA circulation protein, gpP, gp45, gp46, gp47, COG3778, tail fiber protein gp37-C terminal, tail fiber assembly protein, and Com. The last open reading frame was homologous to IS1414. The G + C content of bacteriophage SuMu was 41.87% while its H. parasuis host genome's G + C content was 39.93%. Twenty protein homologs to bacteriophage proteins, including 15 structural proteins, one lysogeny-related and one lysis-related protein, and three DNA replication proteins were identified by mass spectrometry. One of the tail proteins, gp36, may be a virulence-related protein.

CONCLUSIONS

Bacteriophage SuMu was characterized by genomic and proteomic methods and compared to enterobacteriophage Mu.

摘要

背景

猪嗜血杆菌(Haemophilus parasuis)是导致格拉瑟病的病原体,在猪群中广泛存在,与该疾病相关的临床症状包括脑膜炎、多发性浆膜炎、多发性关节炎和细菌性肺炎。在隔离的早期断奶猪群中,6 至 8 周龄的猪特别容易感染该疾病。断奶时初乳抗体不足或在保育或生长-育肥阶段将猪与来自其他农场来源的异源强毒猪嗜血杆菌菌株混合,被认为是爆发格拉瑟病的因素。先前,通过巢式聚合酶链反应在猪嗜血杆菌的一种强毒分离株中检测到 Mu 样噬菌体门户基因。Mu 样噬菌体在系统发育上与肠杆菌噬菌体 Mu 有关,被认为携带毒力基因或诱导宿主表达毒力基因。本研究对从一种强毒猪嗜血杆菌分离株中分离到的 Mu 样噬菌体 SuMu 进行了特征描述。

结果

通过与肠杆菌噬菌体 Mu 的基因组比较和通过质谱法对各种同源物的蛋白质组学鉴定进行了表征。这是首次从猪嗜血杆菌的一种双链 DNA 噬菌体 Mu 样噬菌体中分离和表征来自毒力场分离株的噬菌体,该噬菌体属于肌尾病毒科,具有可收缩的尾巴。噬菌体 SuMu 的基因组大小为 37151bp。DNA 测序显示有 55 个开放阅读框,包括 25 个 Mu 样噬菌体蛋白的同源物:Nlp、噬菌体转座酶-C 末端、COG2842、Gam 样蛋白、gp16、Mor、肽聚糖识别蛋白、gp29、gp30、gpG、gp32、gp34、gp36、gp37、gpL、噬菌体尾管蛋白、DNA 循环蛋白、gpP、gp45、gp46、gp47、COG3778、尾纤维蛋白 gp37-C 末端、尾纤维组装蛋白和 Com。最后一个开放阅读框与 IS1414 同源。噬菌体 SuMu 的 G+C 含量为 41.87%,而其猪嗜血杆菌宿主基因组的 G+C 含量为 39.93%。通过质谱法鉴定了 20 种噬菌体蛋白的同源物,包括 15 种结构蛋白、1 种溶原相关蛋白和 1 种裂解相关蛋白以及 3 种 DNA 复制蛋白。其中一种尾蛋白 gp36 可能是一种与毒力相关的蛋白。

结论

通过基因组和蛋白质组学方法对噬菌体 SuMu 进行了表征,并与肠杆菌噬菌体 Mu 进行了比较。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d399/3447690/c764c9d465eb/1471-2164-13-331-1.jpg

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