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Genomic and Proteomic Analyses of the Terminally Redundant Genome of the Pseudomonas aeruginosa Phage PaP1: Establishment of Genus PaP1-Like Phages

机译:铜绿假单胞菌噬菌体PaP1的末端冗余基因组的基因组和蛋白质组学分析:类PaP1噬菌体的建立。

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摘要

We isolated and characterized a new Pseudomonas aeruginosa myovirus named PaP1. The morphology of this phage was visualized by electron microscopy and its genome sequence and ends were determined. Finally, genomic and proteomic analyses were performed. PaP1 has an icosahedral head with an apex diameter of 68–70 nm and a contractile tail with a length of 138–140 nm. The PaP1 genome is a linear dsDNA molecule containing 91,715 base pairs (bp) with a G+C content of 49.36% and 12 tRNA genes. A strategy to identify the genome ends of PaP1 was designed. The genome has a 1190 bp terminal redundancy. PaP1 has 157 open reading frames (ORFs). Of these, 143 proteins are homologs of known proteins, but only 38 could be functionally identified. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and high-performance liquid chromatography-mass spectrometry allowed identification of 12 ORFs as structural protein coding genes within the PaP1 genome. Comparative genomic analysis indicated that the Pseudomonas aeruginosa phage PaP1, JG004, PAK_P1 and vB_PaeM_C2-10_Ab1 share great similarity. Besides their similar biological characteristics, the phages contain 123 core genes and have very close phylogenetic relationships, which distinguish them from other known phage genera. We therefore propose that these four phages be classified as PaP1-like phages, a new phage genus of Myoviridae that infects Pseudomonas aeruginosa.
机译:我们分离并表征了一种新的铜绿假单胞菌肌病毒,名为PaP1。通过电子显微镜观察该噬菌体的形态,并确定其基因组序列和末端。最后,进行了基因组和蛋白质组学分析。 PaP1的二十面体头的顶直径为68–70 nm,而尾巴的收缩长度为138–140 nm。 PaP1基因组是一个线性dsDNA分子,包含91,715个碱基对(bp),G + C含量为49.36%,并包含12个tRNA基因。设计了识别PaP1基因组末端的策略。基因组具有1190 bp的末端冗余。 PaP1具有157个开放阅读框架(ORF)。其中,143种蛋白质是已知蛋白质的同源物,但在功能上只能鉴定38种。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和高效液相色谱-质谱法鉴定了12个ORF,它们是PaP1基因组内的结构蛋白编码基因。比较基因组分析表明,铜绿假单胞菌噬菌体PaP1,JG004,PAK_P1和vB_PaeM_C2-10_Ab1具有很大的相似性。除了具有相似的生物学特性外,噬菌体还包含123个核心基因,并具有非常紧密的系统发育关系,这使它们与其他已知的噬菌体属区分开来。因此,我们建议将这四个噬菌体归类为PaP1样噬菌体,这是一种感染了铜绿假单胞菌的肌病毒科的新噬菌体属。

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