1株大肠杆菌噬菌体vB_EcoP_GN07的分离鉴定及全基因组分析

doi:10.16431/j.cnki.1671-7236.2023.04.024

Abstract

Abstract: 【Objective】 This study was aimed to explore the biological and genome characteristics of Escherichia coli (E.coli) phage vB_EcoP_GN07,and provide biological materials for the development of phage cocktail formulations against pathogenic E.coli disease.【Method】 A strain of E.coli lytic phage vB_EcoP_GN07 was isolated from a chicken farm in Nanning,Guangxi using the double-layer plate method.Transmission electron microscope observation,host spectrum determination,pH and temperature stability evaluation,the optimal multiplicity of infection (MOI) determination,one-step growth curve determination and whole genome sequencing were used to understand the morphology,biological characteristics and genome-wide characteristics of the phage.【Result】 Transmission electron microscopy showed vB_EcoP_GN07 had a head diameter of about 45 nm±5 nm and tail length of about 25 nm,belonging to the Podoviridae.which had strong host specificity.The phage titer could be kept stable at 4 to 60 ℃,and the phage activity could be maintained at pH 3.0-11.0.The optimal MOI of the phage was 10^-2.The one-step growth curve showed that the incubation period of the phage was 60 min,and the burst size was 560 PFU/cell.Sequencing results showed that it belonged to double stranded DNA,with a length of 70 120 bp and a GC content of 42.8%.The phage vB_EcoP_GN07 had 81 open reading frames,of which 23 were known functional proteins.BLASTN comparison showed that the phage vB_EcoP_GN07 had low similarity with other phages in NCBI database,conformed to the characteristics of N4-like phage,and lacked virulence genes and drug resistance genes associated with antibiotic resistance,toxins and virulence factors.【Conclusion】 An E.coli phage vB_ EcoP_ GN07 was successfully isolated in this study,which had strong cracking ability,host specificity and acid and alkali resistance.vB_ EcoP_ GN07 was less similar to other phages,and determined to be a new N4-like phage.This results provided a theoretical basis for the development of new fungicides with application E.coli phage vB_EcoP_GN07.

Key words: Escherichia coli; phage; biological characteristics; whole genome analysis; comparative genomics

CLC Number:

S852.61⁺2

WANG Leping, PAN Yan, TAN Yizhou, LI Lei, HAN Kaiou, CAO Yajie, LI Jun, ZHOU Yuqing, LIAO Yuying, MA Dongxin, GONG Yu, PENG Hao, WANG Xiaoye. Isolation,Identification and Genome Analysis of Escherichia coli Phage vB_EcoP_GN07[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1522-1531.

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Isolation,Identification and Genome Analysis of Escherichia coli Phage vB_EcoP_GN07

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