铜绿假单胞菌噬菌体D204的生物学特性和基因组学研究

发布时间：2018-01-12 09:40

  本文关键词：铜绿假单胞菌噬菌体D204的生物学特性和基因组学研究　出处：《上海交通大学学报(医学版)》2016年01期 　论文类型：期刊论文

  更多相关文章： 铜绿假单胞菌 噬菌体 基因组 生物学特性

【摘要】：目的针对铜绿假单胞菌多耐药菌的不断出现,寻找高效、广谱的铜绿假单胞菌噬菌体。方法以医院临床上分离到的铜绿假单胞菌为宿主菌,从环境污水中分离纯化铜绿假单胞菌噬菌体,磷钨酸负染法电镜观察噬菌体的形态和大小,测定其效价、宿主谱、酸碱稳定性和热稳定性,绘制一步生长曲线、吸附曲线和感染曲线,并抽取噬菌体DNA进行测序及分析。结果分离到1株铜绿假单胞菌噬菌体,并命名为D204。D204在pH值为4~11及温度37~50℃环境下具有较高活性,电子显微镜观察发现此噬菌体由直径约为50 nm的多面体头部和约15 nm的短尾组成。一步生长曲线表明噬菌体在裂解宿主菌时,潜伏期为30 min,爆发期为40 min,裂解量约为160 pfu/cell。吸附曲线显示50 min时,其吸附率达95%以上。感染曲线显示当感染复数为0.1和1时,对宿主菌生长有抑制作用,在180 min后可完全抑制宿主菌生长。其基因组为环状双链DNA,全长为50 609 bp,GC含量为44.8%。预测共有75个开放阅读框。结论分离、筛选出的铜绿假单胞菌噬菌体D204具有较强的裂解宿主的能力,其对酸碱和热比较稳定,裂解宿主时潜伏期较短,可以作为铜绿假单胞菌的储备噬菌体。
[Abstract]:Objective to search for efficient and broad-spectrum Pseudomonas aeruginosa bacteriophage. Methods Pseudomonas aeruginosa isolated clinically in hospital was used as host bacteria. Pseudomonas aeruginosa phage was isolated and purified from environmental sewage. The morphology and size of phage were observed by phosphotungstic acid negative staining electron microscope. The titer, host spectrum, stability and thermal stability of the bacteriophage were determined, and the one-step growth curve was drawn. The adsorption curve and infection curve were obtained. The phage DNA was sequenced and analyzed. The results showed that a bacteriophage strain of Pseudomonas aeruginosa was isolated. It was named D204.D204 with high activity at pH 4? 11 and temperature 37? 50? 鈩，

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