铜绿假单胞菌噬菌体vB_PaS_IME307的生物学特性研究及全基因组分析

王灿; 刘慧莹; 高明明; 米丽媛; 喻鑫婷; 米志强; 柏长青

doi:10.16016/j.2097-0927.202209112

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铜绿假单胞菌噬菌体vB_PaS_IME307的生物学特性研究及全基因组分析

基础医学 | 更新时间：2026-01-09

- 铜绿假单胞菌噬菌体vB_PaS_IME307的生物学特性研究及全基因组分析
- Biological characteristics and whole genome analysis of Pseudomonas aeruginosa phage vB_PaS_IME307
- 陆军军医大学学报 2023年45卷第19期页码：2045-2056
- 作者机构：
  
  1.236000 安徽阜阳, 安徽医科大学附属阜阳医院呼吸内科
  2.100071 北京, 解放军总医院第五医学中心呼吸与危重症医学科
  3.100071 北京，军事科学院军事医学研究院微生物流行病研究所，病原微生物生物安全国家重点实验室
  4.518000 广东深圳，深圳大学总医院呼吸与危重症医学科
- 作者简介：
  
  [柏长青，E-mail：mlp1604@sina.comBAI Changqing, E-mail: mlp1604@sina.com
- 基金信息：
  
  安徽省自然科学基金青年项目(2108085QH310);安徽省卫生健康委科研项目(AHWJ2021b057);安徽医科大学校科研基金项目(2020xkj060)
- DOI：10.16016/j.2097-0927.202209112
  中图分类号： Q393.48;R378.991;R394.2
- 收稿：2022-09-13，
  
  修回：2023-05-02，
  
  纸质出版：2023-10-15
- 稿件说明：
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王灿, 刘慧莹, 高明明, 等. 铜绿假单胞菌噬菌体vB_PaS_IME307的生物学特性研究及全基因组分析[J]. 陆军军医大学学报, 2023,45(19):2045-2056.

Can WANG, Huiying LIU, Mingming GAO, et al. Biological characteristics and whole genome analysis of Pseudomonas aeruginosa phage vB_PaS_IME307[J]. Journal of Army Medical University, 2023, 45(19): 2045-2056.
王灿, 刘慧莹, 高明明, 等. 铜绿假单胞菌噬菌体vB_PaS_IME307的生物学特性研究及全基因组分析[J]. 陆军军医大学学报, 2023,45(19):2045-2056. DOI： 10.16016/j.2097-0927.202209112.

Can WANG, Huiying LIU, Mingming GAO, et al. Biological characteristics and whole genome analysis of Pseudomonas aeruginosa phage vB_PaS_IME307[J]. Journal of Army Medical University, 2023, 45(19): 2045-2056. DOI： 10.16016/j.2097-0927.202209112.

摘要

目的

以实验室保存菌株Pa175为宿主菌，从医院污水中分离得到1株铜绿假单胞菌噬菌体，研究其生物学特性及全基因组学特征。

方法

利用双层平板法、噬斑法分离并鉴定噬菌体；PEG8000浓缩NaCl沉淀法纯化噬菌体；蔗糖梯度离心进一步纯化噬菌体，经磷钨酸负染染色后，使用透射电子显微镜观察其形态；蛋白酶K/SDS法提取噬菌体DNA，对提取的核酸进行全基因组测序和生物信息学分析。

结果

得到1株裂解性铜绿假单胞菌噬菌体vB_PaS_IME307，该噬菌体可以形成透明且有晕环的噬菌斑，全基因组大小为51 545 bp。透射电镜结果显示vB_PaS_IME307的头部呈正二十面体形状，直径(66±2)nm，尾长(116±2)nm；进化树分析发现该噬菌体为

Mccleskeyvirinae

病毒属，长尾噬菌体科(

Siphoviridae

)。生物学特性研究发现：vB_PaS_IME307在pH= 4.0~11.0、60 ℃以下能够保持较高活性，感染周期约为60 min，其中包括10 min潜伏期和50 min暴发期，其暴发量为150 PFU/cell。

结论

噬菌体vB_PaS_IME307具有宿主特异性，对温度和pH具有较好的耐受能力，具有防治铜绿假单胞菌细菌感染的潜力。

Abstract

Objective

To determine the biological properties and whole genome characteristics of a phage against

P. aeruginosa

isolated from sewage samples as the host strain Pa175.

Methods

Double-layer agar plate and plaque method were used for phage isolation and identification. The obtained phage was purified through PEG 8000 concentration and NaCl precipitation. The phage was further purified with sucrose gradient centrifugation

stained by phosphotungstic acid negative staining. Transmission electron microscopy was employed to observe its morphology. After its DNA was extracted by using proteinase K/SDS

whole genome sequencing was performed and the results were analyzed by bioinformatics.

Results

A lytic

P. aeruginosa

phage was obtained and then named as vB_PaS_IME307

which can form transparent plaque with halo

with a whole genome size of 51 545 bp. Transmission electron microscopy showed that the head of vB_ PaS_ IME307 was icosahedral shape

with a diameter of 66±2 nm and a tail length of 116±2 nm. Phylogenetic tree analysis revealed that IME307 was belonged to Mccleskeyvirinae virus genus

Siphoviridae family. The phage was maintained of high activity at pH value from 4.0 to 11.0 and temperature below 60 ℃. The infection cycle was about 60 min

including 10 min incubation period and 50 min outbreak period. The outbreak was 150 PFU/cell.

Conclusion

vB_ PaS_ IME307 is of host specificity

good tolerance to temperature and pH valule

and has the potential to prevent and treat

P. aeruginosa

infection.

关键词

Keywords

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