产志贺毒素多重耐药大肠杆菌噬菌体的生物学特性及全基因组分析

doi:10.16431/j.cnki.1671-7236.2020.02.005

Abstract

Abstract: To explore the biological characteristics and genome characteristics of Escherichia coli phages,in this study,a Shiga toxin-producing multi-drug resistant Escherichia coli strain GXEC010 was isolated from the sewage of a swine farm in Guangxi,and a lytic phage strain was isolated from the sewage,and through the transmission electron microscope,determination of best multiplicity of infection in the plural,one-step growth curve drawing,thermal sensitivity and pH stability evaluation,sterilization experiment and whole genome sequencing methods to analyze the biological characteristics and genome characteristics of the phage.The results showed that the phage which could efficiently lyse Escherichia coli GXEC010 was isolated and purified,and named as vB_EcoM_BP10.The optimal multiplicity of infection this bacterium was 1.The results of one-step growth curve showed that the incubation period of infected host bacteria was 5 min,the outbreak period was 65 min,and the outbreak quantity was 51.The tolerable temperature range was 30 to 60 ℃,and it could be maintained stable in the pH of 5 to 8.The results of bacteriophage bactericidal experiment showed that the bacteriophage vB_EcoM_BP10 had a good bactericidal effect when MOI=1.Completed genome sequencing results showed that the total fragment length of phage vB_EcoM_BP10 was 52 288 bp,GC content was 44.16%,and included 71 ORF.The GO functional notes indicated that phage vB_EcoM_BP10 could participate in small molecule metabolism,cell nitrogen compound metabolism and other biological processes,and had molecular functions such as nucleotide transferase activity,ion binding and DNA binding.This bacterium was closely related to Escherichia phage ST32 and Enterobacteria phage phiEcoM-GJ1.In conclusion,the phage vB_EcoM_BP10 in this study had clear host receptor specificity,good thermal stability and acid-base stability,which could lay a theoretical foundation for the effective prevention and control of Shiga toxin-producing multidrug-resistant Escherichia coli.

Key words: Shiga toxin; phage; multiple drug resistance; completed genome sequencing

CLC Number:

S852.61

WEI Deyuan, LU Zening, ZHANG Cong, GE Chenling, WANG Xiaoye. A Biological Characteristics and Genome Analysis of Shiga Toxin-producing and Multidrug-resistant Escherichia coli Phages[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(2): 352-362.

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A Biological Characteristics and Genome Analysis of Shiga Toxin-producing and Multidrug-resistant Escherichia coli Phages

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