中国畜牧兽医

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大肠杆菌噬菌体的分离及生物学特性研究

李杨1,2,王静2,霍恺1,李晓颖2,孔健1,谷巍2   

  1. 1.山东大学微生物技术国家重点实验室,山东济南 250100;2.山东宝来利来生物工程股份有限公司,山东泰安 271000)
  • 出版日期:2014-01-20 发布日期:2014-02-27
  • 通讯作者: 孔健(1964—),女,教授,博士生导师,从事微生物学应用基础研究。E-mail: kongjian@sdu.edu.cn
  • 作者简介:李杨(1987—),女,山东人,硕士,从事抗感染微生态研究。
  • 基金资助:

    国家自然基金项目(31271905);国家科技支撑计划项目(2012BAD39B01)。

Isolation and Characterization of Bacteriophage of E.coli Isolated from Soil of Chicken Farm

LI Yang1,2, WANG Jing2, HUO Kai1, LI Xiao-ying2, KONG Jian1, GU Wei2   

  1.   (1.State Key Laboratory of Microbial Technology of Shandong University, Jinan 250100,China;2.Shandong Baolai-Leelai Bioengineering Co., Ltd., Tai’an 271000,China)
  • Online:2014-01-20 Published:2014-02-27

摘要: 试验从鸡场周围环境中采集土样,在伊红美蓝培养基平板上分离纯化大肠杆菌,然后以其为宿主,分离相应的噬菌体,并研究其生物学特性。结果表明,共分离到大肠杆菌4株,经16S rDNA鉴定为埃希氏大肠杆菌。其中,以大肠杆菌LY4为宿主分离纯化到1株烈性噬菌体,负染经电镜观察该噬菌体由多面体头部及尾部组成,噬菌斑直径为2.5~3.8 mm,效价2.0×1010 PFU/mL,最佳感染复数为0.0001;在温度-20~37 ℃、pH 5.0~10.0范围内该噬菌体表现出良好的稳定性,但在人工胃液(pH 1.2)条件下,即可导致死亡;测定其一步生长曲线,该噬菌体潜伏期为20 min,裂解能力强,以上特性表明该噬菌体在鸡舍环境治理中具有潜在的应用价值。

关键词: 大肠杆菌; 噬菌体; 生物学特性

Abstract: 4 of E.coli strains were isolated from the soil of chicken farm on the EMB medium, and further identified as Escherichia coli by 16S rDNA sequence analysis. One virulent phage ΦLY4 was isolated when propagated on the E.coli LY4 as host cells. The phage had the hexagonal head and tail after observing with negatively stained method by electron microscope. The drastic diameter of this phage was 2.5 to 3.8 mm in size, and its phage titer was about 2.0×1010 PFU/mL, the optimal multiplicity of infection was 0.0001. It kept high stability at the temperature of -20 to 37 ℃ and pH at 5.0 to 10.0. Its viability was lost completely at the simulated gastric conditions (pH 1.2). The growth curve showed that its incubation and burst phase was 20 min, and the lysis ability was strong. All of these properties showed that the phage ΦLY4 had a potential application in the control of environmental pollution.

Key words: E.coli; bacteriophage; biological characteristic