Complete Genome Sequence Analysis of Two Pseudomonas plecoglossicida Phages, Potential Therapeutic Agents
Complete Genome Sequence Analysis of Two Pseudomonas plecoglossicida Phages, Potential Therapeutic Agents
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DOI：
10.1128/aem.03038-14
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发表时间：
2015-02-01
影响因子：
4.4
通讯作者：
Nakai, Toshihiro
中科院分区：
文献类型：
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作者：
Kawato, Yasuhiko;Yasuike, Motoshige;Nakai, Toshihiro
Pseudomonas plecoglossicida 是日本香鱼 (Plecoglossus altivelis) 的致命病原体，对香鱼养殖造成巨大的经济损失。之前，我们证明了使用两种裂解性噬菌体（PPpW-3和PPpW-4）进行噬菌体治疗对P. plecoglossicida感染的功效（S. C. Park, I. Shimamura, M. Fukunaga, K. Mori, and T. Nakai, Appl Environ Microbiol 66:1416-1422, 2000, http://dx.doi.org/10.1128/AEM.66.4.1416-1422.2000；S.C.Park 和 T.Nakai，Dis Aquat Org 53:33-39, 2003，http://dx.doi.org/10.3354/dao053033）。在本研究中，确定了这些治疗性 P. plecoglossicida 噬菌体的完整基因组序列，并分析了作为治疗剂的有害因素。 PPpW-3（肌病毒）基因组由43,564 bp组成，GC含量为61.1%，有66个预测开放阅读框（ORF）。大约一半的基因与大肠杆菌噬菌体vB_EcoM_ECO1230-10（肌病毒）的基因相似。 PPpW-4（足病毒）的基因组由 41,386 bp 组成，GC 含量为 56.8%，预测 ORF 50 个。超过70%的基因与荧光假单胞菌噬菌体phi IBB-PF7A和恶臭假单胞菌噬菌体phi 15（足病毒）的基因相似。全基因组分析显示，PPpW-3 和 PPpW-4 中不存在已知的毒力基因。在 PPpW-3 中发现了整合酶基因，但用于溶源的其他因子尚未得到证实。对噬菌体抗性变体中噬菌体基因的 PCR 检测没有提供 PPpW-3 和 PPpW-4 中溶原活性的证据。我们得出的结论是，这两种裂解噬菌体有资格作为治疗剂。
Pseudomonas plecoglossicida is a lethal pathogen of ayu (Plecoglossus altivelis) in Japan and is responsible for substantial economic costs to ayu culture. Previously, we demonstrated the efficacy of phage therapy against P. plecoglossicida infection using two lytic phages (PPpW-3 and PPpW-4) (S. C. Park, I. Shimamura, M. Fukunaga, K. Mori, and T. Nakai, Appl Environ Microbiol 66:1416-1422, 2000, http://dx.doi.org/10.1128/AEM.66.4.1416-1422.2000; S. C. Park and T. Nakai, Dis Aquat Org 53:33-39, 2003, http://dx.doi.org/10.3354/dao053033). In the present study, the complete genome sequences of these therapeutic P. plecoglossicida phages were determined and analyzed for deleterious factors as therapeutic agents. The genome of PPpW-3 (myovirus) consisted of 43,564 bp with a GC content of 61.1% and 66 predicted open reading frames (ORFs). Approximately half of the genes were similar to the genes of the Escherichia coli phage vB_EcoM_ECO1230-10 (myovirus). The genome of PPpW-4 (podovirus) consisted of 41,386 bp with a GC content of 56.8% and 50 predicted ORFs. More than 70% of the genes were similar to the genes of Pseudomonas fluorescens phage phi IBB-PF7A and Pseudomonas putida phage phi 15 (podoviruses). The whole-genome analysis revealed that no known virulence genes were present in PPpW-3 and PPpW-4. An integrase gene was found in PPpW-3, but other factors used for lysogeny were not confirmed. The PCR detection of phage genes in phage-resistant variants provided no evidence of lysogenic activity in PPpW-3 and PPpW-4. We conclude that these two lytic phages qualify as therapeutic agents.