一株裂解性奶牛乳房炎源大肠杆菌噬菌体的分离鉴定及生物学特性分析

发布时间：2019-01-30 12:48

【摘要】：【目的】从新疆石河子地区奶牛粪样中分离裂解性大肠杆菌噬菌体(Escherichia coli phage),对其进行纯化及生物学特性分析。【方法】利用双层平板法从奶牛粪样中分离、纯化噬菌体,将纯化后的噬菌体浓缩液用醋酸双氧铀负染后通过透射电子显微镜观察其形态特征。对该噬菌体进行全基因组测序和遗传进化分析,同时测定噬菌体的宿主谱、最佳感染复数、一步生长曲线、热稳定性及酸碱稳定性。【结果】分离并纯化出一株裂解性噬菌体vB_EcoM_XJ2,噬菌斑圆形不透明,直径0.7 mm 1.2 mm;电镜显示其头部呈正多面体对称,有可伸缩性尾部;核酸类型为双链DNA,基因组大小为75.617 kb,G+C%含量为42.09%;其核酸序列与大肠杆菌噬菌体NJ01和vB_EcoP_SU10相似性高达94%。生物学特性研究显示该噬菌体能裂解多株临床分离的大肠杆菌;能耐受60°C左右高温,在pH 5.0 11.0范围内效价稳定;最佳感染复数为0.1,潜伏期为15 min,暴发期为95 min,裂解量约为10.6 PFU/cell。【结论】vB_EcoM_XJ2是一株在不同温度、不同酸碱性环境中有较强适应能力的裂解性肌尾科大肠杆菌噬菌体。
[Abstract]:[objective] to purify lytic Escherichia coli phage (Escherichia coli phage), from cow dung samples in Shihezi area, Xinjiang, and to analyze its biological characteristics. [methods] Phage phage was isolated and purified from cow dung samples by double plate method. The purified bacteriophage concentrate was negatively stained with dioxic uranium acetate and its morphological characteristics were observed by transmission electron microscope (TEM). The whole genome of the phage was sequenced and the genetic evolution was analyzed. At the same time, the host spectrum of the phage was determined. Thermal stability and acid-base stability. [results] A lytic bacteriophage vB_EcoM_XJ2, phagocytic plaque was isolated and purified with circular opacity with a diameter of 0.7 mm / 1. 2 mm;. Electron microscope showed that its head was regular polyhedron and had scalable tail, and the nucleic acid type was 75.617 kb,G C% of double-stranded DNA, genome, and 42.09% of nucleic acid. The nucleic acid sequence was similar to that of Escherichia coli phage NJ01 and vB_EcoP_SU10. The biological characteristics of the bacteriophage showed that the bacteriophage could cleavage many clinical isolates of Escherichia coli, and could withstand high temperature of about 60 掳C, and its titer was stable in the range of pH 5.0 ~ 11.0. The best complex number of infection was 0.1, the incubation period was 15 min, and the 95 min, cleavage was about 10.6 PFU/cell. [conclusion] vB_EcoM_XJ2 was a strain at different temperatures. Lytic Escherichia coli phage with strong adaptive ability in different acid-base environments.
【作者单位】：石河子大学动物科技学院;军事医学科学院微生物流行病研究所;新疆生产建设兵团畜牧兽医工作总站;
【基金】：国家自然科学基金项目(No.31560699) 新疆生产建设兵团应用基础研究计划项目(No.2015AG012)~~
【分类号】：S852.6

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