一株副溶血弧菌噬菌体的分离鉴定及生理特性

Isolation, identification and physiological property of lytic phage against Vibrio parahaemolyticus

摘要:

为了探寻有效的控制副溶血弧菌的方法, 从水产品市场的污水中分离出来一株烈性噬菌体qdvp001。采用双层平板法分离纯化噬菌体qdvp001, 电镜观察其形态特征, 并利用双层平板法测定其生理特性, 包括热稳定性试验、最适pH、最佳感染复数及一步生长曲线, 然后提取基因组进行酶切和序列分析。结果显示该烈性噬菌体qdvp001头部直径大约为79 nm, 尾长大约118 nm, 属于肌尾噬菌体科。它对60 °C以下的温度耐受力较强, 最适pH为7.0?8.0左右, 最佳感染复数是0.000 1, 感染宿主菌的潜伏期约20 min, 裂解期约70 min, 并获得部分DNA片段的序列。将获得的DNA序列在NCBI上进行比对, 结果显示, qdvp001与其他噬菌体的同源性较低。该噬菌体很可能是一种新发现的噬菌体。

Abstract:

To develop a effective controlling method against Vibrio parahaemolyticus, we isolated a lytic phage qdvp001 from aquatic sewage water. Phage qdbp001 was isolated by double-layer plate method and its morphology character were observed by transmission electron microscopic. Furthermore some physiological properties such pH stability, thermal stability, multiplicity of infection (MOI) and one step curve were also reported. Deoxyribose nucleic acid (DNA) from phage qdbp001 was digested with the restriction endonuclease and then analyzed the sequence. Phage qdvp001 with a head diameter of about 79 nm and a tail size of about 102 nm belongs to the family Myoviridae. It has a strong endurance of temperature below 60 °C, its optimum pH value and MOI were about 7.0?8.0 and 0.000 1 re-spectively. The phage growth cycle with a detected latent period of 20 min with lytic period of 10 min. Six DNA sequences were obtained and showed low homology with others when compared with those reported information in NCBI. Phage qdvp001 might be a novel bacteriophage.