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目的:以14株阴沟肠杆菌为宿主菌,从环境污水中分离噬菌体,分析其生物学特性,鉴定其分型。方法:采用双层琼脂噬斑法分离并鉴定噬菌体。纯化后的噬菌体经负染法染色,通过电镜观察其大小和形态;提取噬菌体基因组,进行酶切电泳分析;通过裂解谱的分析,确认噬菌体的特异性和裂解宿主菌范围。结果:成功分离1株裂解性噬菌体,电镜显示噬菌体的头部呈球形,直径约60nm,尾部长约130nm。琼脂糖凝胶电泳显示其基因组约40 000bp,且含多种酶切位点。特异性实验显示其呈现较窄的宿主范围。结论:分离的阴沟肠杆菌噬菌体(命名为ФEc53)属于有尾病毒目,管尾病毒科,是一种特异性高、裂解性强的毒性噬菌体。
OBJECTIVE: To isolate 14 phage Enterobacter cloacae bacteria from environmental wastewater and analyze its biological characteristics and identify its type. Methods: Two-layer agar plaque was used to isolate and identify phage. The purified phage was stained by negative staining and the size and morphology of the phage were observed by electron microscopy. The phage genome was extracted and analyzed by restriction endonuclease analysis. The specificity of the phage and the range of lytic host bacteria were confirmed by analysis of the cleavage spectrum. Results: A lytic bacteriophage was successfully isolated. Electron microscopy showed that the phage had a spherical head with a diameter of about 60 nm and a tail length of about 130 nm. Agarose gel electrophoresis showed that the genome of about 40 000bp, and contains a variety of enzyme cutting sites. Specific experiments show that it presents a narrower host range. Conclusion: The isolated Enterobacter cloacae phage (designated as ФEc53) belongs to the order of the causative virus and the family of terminal virus. It is a highly specific and highly lytic bacteriophage.