一株宽裂解谱鼠疫噬菌体的生物特性和基因组研究

靳海晓; 钟佑宏; 郭瑾; 张传宇; 李论; 申小娜; 段存娟; 黄英; 熊浩明; 王鹏; 李伟

doi:10.3784/jbjc.202212130534

一株宽裂解谱鼠疫噬菌体的生物特性和基因组研究

Biological and genomic characteristics of a broad-spectrum Yersinia pestis bacteriophage

摘要

摘要:
目的鉴定一株分离自既往鼠疫疫源地的褐家鼠盲肠的鼠疫噬菌体vB_YpeM_ESS14（ESS14）的生物学特性、电镜形态及其基因组特征。
方法以鼠疫疫苗株EV76为宿主菌，通过测定噬菌体ESS14的裂解谱、最佳感染复数、一步生长曲线、热稳定性、PH稳定性和紫外线稳定性以及透射电子显微镜形态观察了解ESS14的生物学特性，通过基因组测序分析阐述其基因组特征。
结果 ESS14能裂解多种大肠埃希菌和志贺菌，此外ESS14还能裂解鼠伤寒沙门菌和布利丹沙门菌；ESS14的最佳感染复数为0.01，爆发潜伏期10 min，爆发量为120 PFU/ml；ESS14在温度28～60 ℃ 和pH （3～10）条件下效价稳定；ESS14基因组由170 013 bp的双链DNA组成，G+C含量为40.50%，包含295个开放阅读框（ORFs），其中有2个为tRNA （tRNA-Arg-TCT和tRNA-Met-CAT）。
结论基因组特定鉴定分析表明ESS14属于Caudovirales>Straboviridae>Tevenvirinae>Gapriverviru；ESS14具有较宽的裂解谱，潜伏期短，爆发量高，对温度、pH、紫外线稳定性良好，未来用其作微生态制剂减灭鼠疫疫源地中的鼠疫菌方面具有潜在的应用价值。

Abstract:
Objective To determined the morphological, biological and genomic characteristics of vB_YpeM_ESS14 (ESS14), a Yersinia pestis bacteriophage isolated from a silent plague focus in Yunnan.
Methods Using Y.pestis vaccine strain EV76 as host bacteria, the morphological, biological and genomic characteristics of ESS14 were determined by transmission electron microscope, growth and thermo, pH stability tests, as well as host range analysis.
Results ESS14 could lyse some species of Escherichia coli and Shigella, in addition ESS14 can lyse Salmonella typhimurium and Salmonella blidan. The Optimal Multiplicity of Infection (MOI) of ESS14 was 0.01, the latent period was shorter, and the burst size was about 120 PFU/cell. ESS14 has comparatively stability at temperatures range (28～60 ℃) and pHs range (3～10). The ESS14 genome consists of 170013 bp double stranded DNA, with a G+C content of 40.50%, and contains 295 coding sequences(CDS), two of which are tRNAs (tRNA-Arg-TCT and tRNA-Met-CAT).
Conclusion Genomic analysis revealed that ESS14 belonged to Caudovirales>Straboviridae>Tevenvirinae>Gaprivervirus. Because this phage has the characteristics of broad-lysis spectrum, short latent period, high burst size, as well as the ability with stability to temperature, pH and UV, such characteristics indicated that ESS14 has the application value on reducing Y.pestis in plague focus in the future.

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