1株C3样奇异变形杆菌烈性噬菌体的生物学特性及基因组分析

doi:10.11843/j.issn.0366-6964.2021.04.022

摘要/Abstract

摘要： 从仔猪粪样中分离鉴定奇异变形杆菌（Proteus mirabilis）的烈性噬菌体，并对其进行生物学特性分析和基因组分析。以Proteus mirabilis为宿主菌，用双层平板法从仔猪粪样中分离噬菌体。利用电子显微镜观察噬菌体大小和形态。提取噬菌体DNA，并使用Illumina Hiseq进行高通量测序，对基因组进行注释和分析。结果显示：分离到的奇异变形杆菌噬菌体命名为vB_PmiP_P59（Proteus mirabilis phage vB_PmiP_P59）。P59属于短尾噬菌体科，形态为C3形：头部为扁平的椭圆形，长152 nm，宽54 nm；尾部长约40 nm，宽12 nm。P59的最佳感染复数为0.001；潜伏期为30 min，爆发期为105 min，裂解量为2 754 PFU·cell^-1；P59在不高于50℃，pH 5~10时稳定。P59噬菌体基因组全长90 187 bp，（G+C）%含量为34.65%，共编码136个ORF和4个tRNA，不携带抗生素抗性基因和毒力基因。蛋白质组分析表明，P59与C3样噬菌体phiEco32有30.47%的一致性。奇异变形杆菌噬菌体vB_PmiP_P59属于C3样烈性噬菌体，并对奇异变形杆菌有显著的抑菌效果。

关键词: 奇异变形杆菌, 噬菌体, C3, 生物学特性, 基因组学

Abstract: This study was conducted to isolate and characterize a lytic bacteriophage against Proteus mirabilis from piglet fecal samples. Bacteriophage was isolated and purified with indicator of Proteus mirabilis by double-layer agar plate method from fecal samples of piglets. The size and shape of phage was observed with transmission electron microscopy. The whole genome of phage was extracted and sequenced by Illumina Hiseq high-throughput sequencing technology, and then the genome was annotated and analyzed. A lytic phage (designated as vB_PmiP_P59) against Proteus mirabilis was successfully isolated. P59 was a podovirus with C3 morphotype: a flattened oval head of 152 nm×54 nm and a tail of 40 nm×12 nm. The optimal MOI of P59 was 0.001.The one-step growth curve showed that P59 had a latent period of 30 min, a burst period of 105 min, and a burst size of 2 754 PFU·cell^-1. The phage could withstand the temperature up to 50 ℃ and maintain stable titer under pH 5-10.The double-stranded DNA genome was 90 187 bp long with a mol% G+C content of 34.65, and contained 136 ORFs and 4 tRNAs without any virulent or antibiotic resistance genes. Whole-genome proteomic analysis (30.47% homologous proteins) revealed that it was related to C3-like phage phiEco32. A novel lytic Proteus mirabilis phage vB_PmiP_P59 is identified with significant ability to inhibit the growth of Proteus mirabilis.

Key words: Proteus mirabilis, bacteriophage, C3, biological characterization, genomic analysis

中图分类号:

Q939.48

李淑雅, 黄赞, 朱伟云, 林焱. 1株C3样奇异变形杆菌烈性噬菌体的生物学特性及基因组分析[J]. 畜牧兽医学报, 2021, 52(4): 1069-1078.

LI Shuya, HUANG Zan, ZHU Weiyun, LIN Yan. Biological Characteristics and Genomic Analysis of a Lytic Proteus mirabilis Phage with C3 Morphotype[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(4): 1069-1078.

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