Isolation and identification of phage SWH-Ab-1 against <italic style="font-style: italic">Acinetobacter baumannii</italic> and bioinformatic analysis of its major functional genes

Juan LUO; Ran ZHANG; Liu WU; Xi LUO; Bo ZHANG

doi:10.16016/j.1000-5404.201708010

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Isolation and identification of phage SWH-Ab-1 against Acinetobacter baumannii and bioinformatic analysis of its major functional genes

Basic Medicine | 更新时间：2026-01-09

- Isolation and identification of phage SWH-Ab-1 against Acinetobacter baumannii and bioinformatic analysis of its major functional genes
- Journal of Third Military Medical University Vol. 40, Issue 1, Pages: 23-30(2018)
- 作者机构：
  
  1.400038 重庆, 陆军军医大学西南医院感染管理科
  2.330031 南昌, 南昌大学医学院玛丽女王学院中英2014级6班
- 作者简介：
- 基金信息：
  
  the Special Project of Clinical Research of the First Affiliated Hospital of Third Military Medical University(SWH2014LC31);the Project of Technology Innovation of Social People's Livelihood of Chongqing(cstc2015shmszx120110);the Science and Technology Development Foundation of Engineering Physics Institute of China (Cross Direction of Physics and Biomedicine)(WSS-2015-01)
- DOI：10.16016/j.1000-5404.201708010
  CLC： Q755;Q782;R378.79
- Received：02 August 2017，
  
  Revised：2017-12-05，
  
  Published：15 January 2018
- 稿件说明：
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Juan LUO, Ran ZHANG, Liu WU, et al. Isolation and identification of phage SWH-Ab-1 against Acinetobacter baumannii and bioinformatic analysis of its major functional genes[J]. Journal of Third Military Medical University, 2018, 40(1): 23-30.
DOI：

Juan LUO, Ran ZHANG, Liu WU, et al. Isolation and identification of phage SWH-Ab-1 against Acinetobacter baumannii and bioinformatic analysis of its major functional genes[J]. Journal of Third Military Medical University, 2018, 40(1): 23-30. DOI： 10.16016/j.1000-5404.201708010.

摘要

目的

分离鲍曼不动杆菌噬菌体

测定其基因组DNA序列

分析预测穿孔素和裂解酶等重要功能基因及其编码蛋白的性质和功能。

方法

利用双层琼脂平板法从医院污水中分离鲍曼不动杆菌噬菌体

透射电子显微镜观察噬菌体的形态特征; 提取噬菌体基因组DNA

采用Illumina Hiseq2000测序仪进行全基因组测序; 利用生物信息学方法对噬菌体基因组DNA序列进行功能编码基因定位、同源性分析以及对噬菌体重要功能基因编码蛋白的结构和生物功能等进行预测。

结果

成功从医院污水中分离获得1株鲍曼不动杆菌噬菌体

命名为噬菌体SWH-Ab-1。噬菌体SWH-Ab-1属长尾噬菌体

对鲍曼不动杆菌具有显著裂解性。噬菌体SWH-Ab-1基因组为全长41 567 bp的双链DNA

G+C%含量为39.42%

包含51个预测功能基因

穿孔素基因和裂解酶基因分别位于噬菌体基因组的ORF02(320~655 bp)和ORF03(642~1 199 bp)开放阅读框

穿孔素基因序列全长336 bp

共编码111个氨基酸

其编码的HolSA1为疏水性Ⅰ型跨膜蛋白

相对分子质量为11.957 96×10

理论等电点为4.87

可能在脂肪酸代谢、转运载体及信号转导中发挥重要作用。裂解酶基因序列全长558 bp

共编码185个氨基酸

其编码的LysSA1为亲水性蛋白

相对分子质量为21.010 4×10

理论等电点为9.56

无跨膜区

可能参与氨基酸生物合成、脂肪酸代谢、辅因子生物合成等重要生理过程。两种功能蛋白均不存在信号肽序列

二级结构主要以α-螺旋和无规则卷曲为主。

结论

噬菌体SWH-Ab-1是一种新分离的长尾裂解性噬菌体

其基因组的ORF02和ORF03开放阅读框是穿孔素和裂解酶的编码基因

编码的HolSA1和LysSA1可能组成“二元裂解系统”

协同参与噬菌体对宿主菌的裂解过程。

Abstract

Objective

To isolate the

Acinetobacter baumannii

(

A.baumannii

) phages

sequence the genome and predict the properties and functions of the major functional genes such as holin and lyase.

Methods

A.baumannii

phage was isolated from raw sewage of a hospital and identified using double-layer-agar method.The morphology of the phage was observed by electron microscopy.The genomic DNA of the phage was extracted and sequenced using Illumina Hiseq2000 technique.Bioinformatic analysis was used to analyze the nucleotide sequence of the phages

including functional genes location and homology analysis

and to predict the structures and biological functions of the major encoding proteins.

Results

A.baumannii

phage was successfully isolated and named as SWH-Ab-1.Bacteriophage SWH-Ab-1 belonged to the

Siphoviridae

which had significant lytic activity against

A.baumannii

.The genome of bacteriophage SWH-Ab-1 was a double-strand DNA with a full length of 41 567 bp

in which the G+C mol% was 39.4% and 51 predicted genes were involved.The holin gene and the lysin gene were located in the ORF02 (320~655 bp) and the ORF03 (642~1 199 bp) of SWH-Ab-1

respectively.HolSA1

consisting of 111 amino acids (336 bp)

was a hydrophobic Ⅰ transmembrane protein

at a molecular mass of 11.957 96×10

with a theoretical isoelectric point of 4.87

which might play an important role in fatty acid metabolism

transport and signal transduction.LysSA1 consisted of 185 amino acids (558 bp)

was a hydrophilic protein without transmembrane regions

its molecular mass was 21.010 4×10

and its theoretica

l isoelectric point was 9.56

which might function in amino acid biosynthesis

fatty acid metabolism and biosynthesis of cofactors.The secondary structure of the 2 proteins was mainly composed of alpha helixes and random coil

and both of them were lack of signal peptide.

Conclusion

Bacteriophage SWH-Ab-1 is a new lytic phage

belonging to the

Siphoviridae

family.The open reading frames ORF02 and ORF03 are the encoding genes of phage SWH-Ab-1 genome.HolSA1 and LysSA1 might form a "two-element lysis system" together in order to lyse the host bacteria.

关键词

Keywords

references

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Isolation and identification of phage SWH-Ab-1 against Acinetobacter baumannii and bioinformatic analysis of its major functional genes

DOI：10.16016/j.1000-5404.201708010

摘要

Abstract

关键词

Keywords

references