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粪肠球菌裂解性噬菌体φEn-ZZ8生物学特性的初步分析
张哲,李菁华,逯茵茵,韩放,李诗恒,孙延波
吉林大学基础医学院 病原生物学系,吉林 130021
Biological characteristics of a virulent bacteriophage φEn­-ZZ8 isolated from Enterococcus faecalis
ZHANG Zhe, LI Jing-Hua, LU Yin-Yin, HAN Fang, LI Shi-Heng, SUN Yan-Bo
Department of Pathogenobiology, Basic Medical School of Jilin University, Changchun, 130021, China
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摘要: 本研究旨在通过分离粪肠球菌裂解性噬菌体并分析其生物学特性,为噬菌体疗法应用于肠球菌感染提供依据。以粪肠球菌为宿主菌,用双层琼脂法从环境污水中分离出1株裂解性噬菌体(命名为φEn-­ZZ8),对其生物学特性进行初步分析。透射电镜观察发现,φEn­-ZZ8头部呈二十面体立体对称,直径约40 nm;尾部呈丝状,长约200 nm。吸附实验表明,在10 min内超过90%的噬菌体可吸附至宿主菌细胞。一步生长曲线显示,φEn­-ZZ8的潜伏期为15 min,爆发期为40 min,爆发量为150 pfu/cell。φEn­-ZZ8基因组对内切酶Hind Ⅲ、EcoR Ⅴ、NdeⅠ、PstⅠ、XbaⅠ和SacⅡ敏感;基因组大小约为42 kb。十二烷基硫酸钠­聚丙烯酰胺凝胶电泳至少可见14条蛋白带,相对分子质量为10 000~130 000。φEn­-ZZ8对26株粪肠球菌的裂解率达30.8%,其裂解活性在pH 5~8和50~70 ℃条件下仍稳定。结果提示,φEn­-ZZ8是一种裂解性强、裂解谱较宽、对温度具有耐受性的裂解性噬菌体,有望成为抗耐药性肠球菌感染的噬菌体生物制剂。

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张哲
李菁华
逯茵茵
韩放
李诗恒
孙延波
关键词 粪肠球菌耐万古霉素肠球菌噬菌体噬菌体疗法    
Abstract:This study aims to isolate lytic bacteriophages from Enterococcus faecalis (E. faecalis),analyze its biological characteristics and to establish the research basis for a phage­based therapy against Enterococcus infections. A lytic bacteriophage (named φEn­-ZZ8) was isolated from the sewage using double­agar method and its basic biological characteristics were analyzed. The results showed that φEn­-ZZ8 had an icosahedral head of 40 nm in diameter and a filamentous tail of 200 nm in length. The adsorption rate of φEn-­ZZ8 reached 90% in 10 min. One­step growth showed that φEn­-ZZ8 had a short latency period of 15 min, a rise period of 40 min, and a burst size of 150 pfu/cell. The genome of φEn­-ZZ8 was sensitive to HindIII, EcoRV, NdeI, PstI, Xba I and Sac II, and the size of the genome was estimated to be 42 kb. Sodium dodecyl sulfate­polyacrylamide gel electrophoresis (SDS­-PAGE) revealed that at least 14 proteins were observed on the gel, ranging from 11 000 to 130 000. The bacteriophage specifically infected and lysed 30.8% E. faecalis isolates. The φEn­-ZZ8 also exhibited stability to the wide range of pH and temperature. The data suggest that phage φEn-­ZZ8 may be a candidate therapeutic agent to control Enterococcus infections.

Key wordsEnterococcus faecalis    Vancomycin-resistant Enterococcus    bacteiophage    Phage therapy
基金资助:吉林省科技厅科技发展计划国际科技合作项目(20140414042GH)

通讯作者: 孙延波   
引用本文:   
张哲,李菁华,逯茵茵,韩放,李诗恒,孙延波. 粪肠球菌裂解性噬菌体φEn-ZZ8生物学特性的初步分析[J]. 微生物与感染, 2015, 10(4): 241-246.
ZHANG Zhe, LI Jing-Hua, LU Yin-Yin, HAN Fang, LI Shi-Heng, SUN Yan-Bo. Biological characteristics of a virulent bacteriophage φEn­-ZZ8 isolated from Enterococcus faecalis. JOURNAL OF MICROBES AND INFECTIONS, 2015, 10(4): 241-246.
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http://jmi.fudan.edu.cn/CN/     或     http://jmi.fudan.edu.cn/CN/Y2015/V10/I4/241

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