| 特异性识别K1荚膜大肠杆菌的噬菌体PNJ1809-36生物学特性及全基因组分析 |
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| 引用本文： | 巩倩雯,李一昊,曾颃,于沛欣,钱新杰,王瑜欣,戴建君,汤芳. 特异性识别K1荚膜大肠杆菌的噬菌体PNJ1809-36生物学特性及全基因组分析[J]. 畜牧兽医学报, 2021, 52(6): 1677-1688. DOI: 10.11843/j.issn.0366-6964.2021.06.022 |
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| 作者姓名： | 巩倩雯 李一昊 曾颃 于沛欣 钱新杰 王瑜欣 戴建君 汤芳 |
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| 作者单位： | 南京农业大学动物医学院 教育部动物健康与食品安全国际联合实验室 农业农村部细菌学重点实验室, 南京 210095 |
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| 基金项目： | 国家重点研发计划（2019YFC1605400）；江苏省自然科学基金优秀青年基金项目（BK20180075） |
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| 摘 要： | 本研究以K1荚膜大肠杆菌DE058(avian pathogenic Escherichia coli)为宿主菌,从鸡粪中分离烈性噬菌体PNJ1809-36并对其进行生物学特性的研究和基因组测序及分析.将噬菌体PNJ1809-36进行分离纯化,通过形态学观察、最佳MOI测定、一步生长曲线测定、温度和酸碱耐受性测定、体外...
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| 关 键 词： | 噬菌体 K1荚膜 大肠杆菌 生物学特性 全基因组分析 |
| 收稿时间： | 2020-09-22 |
Biological Characteristics and Whole Genome Analysis of Phage PNJ1809-36 Target Escherichia coli K1 |
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| GONG Qianwen,LI Yihao,ZENG Hang,YU Peixin,QIAN Xinjie,WANG Yuxin,DAI Jianjun,TANG Fang. Biological Characteristics and Whole Genome Analysis of Phage PNJ1809-36 Target Escherichia coli K1[J]. Chinese Journal of Animal and Veterinary Sciences, 2021, 52(6): 1677-1688. DOI: 10.11843/j.issn.0366-6964.2021.06.022 |
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| Authors: | GONG Qianwen LI Yihao ZENG Hang YU Peixin QIAN Xinjie WANG Yuxin DAI Jianjun TANG Fang |
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| Affiliation: | MOE Joint International Research Laboratory of Animal Health and Food Safety/Key Laboratory of Animal Bacteriology, Ministry of Agriculture and Rural Affairs/College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China |
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| Abstract: | Taking Escherichia coli (E. coli) K1 strain DE058 as a host, a virulent phage PNJ1809-36 was isolated from chicken feces. The biological characteristics and genome analysis of this phage were studied. Phage PNJ1809-36 was isolated and purified, whereafter, the biological characteristics of the phage were studied by analyzing the morphology, optimal MOI, one-step growth curve, temperature and acid-base tolerance, and lysis curve in vitro. Moreover, the whole genome of the phage was sequenced and analyzed. The results showed that phage PNJ1809-36 formed transparent plaque with a clear boundary. Under microscopy, phage PNJ1809-36 presented a contractile tail, indicating that it belongs to Myoviridae. The result of host range test showed that phage PNJ1809-36 specifically targeted E. coli with K1 capsule. The optimal MOI of phage PNJ1809-36 was 0.01. One-step growth curve showed that the latent and explosive period was 10 min and 40 min, respectively. Phage was stable at 40 and 50℃, while lost activity at 60℃ for 30 min. Phage PNJ1809-36 could maintain activity under conditions of pH 3-11, with the optimal pH 6; The lysis curve showed that the phage could continuously inhibit the growth of host bacteria for 6 h. Genomic analysis showed that it had low sequence identity with early reported E. coli K1-specific phage, such as K1F and K1E, but presented over 90% sequence homology with newly discovered E. coli K1-specific phage at nucleic acid level, such as nepoznato, PVP-SE1, and phAPEC8. PNJ1809-36, a lytic phage targeting E. coli K1, belongs to a new phylogenetic branch of Myoviridae. Due to the specificity to E.coli K1, phage PNJ1809-36 has the potential to identify and control the diseases caused by E. coli K1. |
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| Keywords: | phage K1 capsule Escherichia coli biological characteristic genome-wide analysis |
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