腾冲热海一株栖热菌裂解性噬菌体的分离及其特征

Isolation and characterization of a Thermus lytic bacteriophage from Tengchong Rehai hot spring

摘要:

摘要: 【目的】Thermus（栖热菌）属是嗜热细菌中比较古老的类群，本研究探索从腾冲热海热泉分离栖热菌噬菌体，并初步分析其特征。【方法】采用“双层平板法”从云南腾冲热海碱性热泉中分离纯化栖热菌噬菌体；对噬菌体及其宿主进行电镜形态观察，并进行噬菌体基因组限制性酶切片段多态性分析、噬菌体生理特征及蛋白组成分析。【结果】从腾冲热海热泉分离获得1株裂解性噬菌体，其宿主菌TC10通过16S rRNA基因序列分析鉴定为Thermus属菌株。此噬菌体为长尾型，头部直径67 nm，尾管长837 nm、宽10 nm，最适感染温度为65-70℃，最适感染pH值为7.6，对氯仿不敏感，其形态与分离自俄罗斯勘察加半岛热泉的栖热菌噬菌体P23-45和P74-26有一定的差异，蛋白组成差异显著，为一株新的栖热菌噬菌体，命名为TTSP10（Tengchong Thermus Siphoviridae Bacteriophage）。

Abstract:

Abstract:[Objective] Genus Thermus represents an ancient descendant within the domain of Bacteria. This research was focused on the isolation and characteristics of Thermus bacteriophages from Tengchong Reihai hot spring. [Methods] Bacteriophage was isolated from Tengchong Rehai hot springs by double-layer plate method, and further characterized by morphology, temperature, pH and organic solvent effection on phage production, DNA restriction endonuclease digestion and protein composition analysis.[Results] One lytic bacteriophage was isolated from Tengchong hot spring. It’s host strain Thermus sp.TC10 belonged to genus Thermus (16S rRNA gene accession number GU119889). This phage has a hexagonal head (67 nm in diameter) and an extremely long tail (837 nm in length and 10 nm in width). The optimum temperature and pH value for production of virons were about 65℃ and pH7.6, respectively. The phage was not sensitive to chloroform. The differences between this phage and the other two Thermus Siphoviridae phages P23-45 and P74-26, which were isolated form Russia's Kamchatka peninsula, demostrated it was a novel bacteriaphage and was denoted as TTSP10 (Tengchong Thermus Siphoviridae phage).