Department of Microbiology, College of Basic Medical Science, Third Military Medical University, Chongqing, China.
PLoS One. 2013 May 13;8(5):e62933. doi: 10.1371/journal.pone.0062933. Print 2013.
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.
我们分离并鉴定了一种新的铜绿假单胞菌噬菌体，命名为 PaP1。该噬菌体的形态通过电子显微镜观察，其基因组序列和末端也已确定。最后，进行了基因组和蛋白质组分析。PaP1 具有一个直径为 68-70nm 的二十面体头部和一个长度为 138-140nm 的收缩尾部。PaP1 基因组是一个线性 dsDNA 分子，包含 91715 个碱基对 (bp)，G+C 含量为 49.36%，含有 12 个 tRNA 基因。设计了一种鉴定 PaP1 基因组末端的策略。基因组具有 1190bp 的末端冗余。PaP1 有 157 个开放阅读框 (ORFs)。其中，143 个蛋白是已知蛋白的同源物，但只有 38 个可以被功能鉴定。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和高效液相色谱-质谱联用允许鉴定 PaP1 基因组中的 12 个 ORF 为结构蛋白编码基因。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 具有高度相似性。除了具有相似的生物学特性外，这些噬菌体还含有 123 个核心基因，具有非常密切的系统发育关系，这使它们与其他已知噬菌体属区分开来。因此，我们建议将这四种噬菌体归类为 PaP1 样噬菌体，这是一种感染铜绿假单胞菌的肌尾噬菌体科的新噬菌体属。
