微小单孢菌噬菌体PhiHAU8的分离、特性鉴定及构建成噬菌粒
Isolation and characterization of Micromonospora phage PhiHAU8 and development into a phasmid.
作者信息
Li Xiaohua, Zhou Xiufen, Deng Zixin
机构信息
Bio-X Life Science Research Center, Shanghai Jiaotong University, Shanghai 200030, People's Republic of China.
出版信息
Appl Environ Microbiol. 2004 Jul;70(7):3893-7. doi: 10.1128/AEM.70.7.3893-3897.2004.
PhiHAU8, a temperate Micromonospora phage, which is capable of infecting Micromonospora sp. strains 40027 and A-M-01, was isolated. The PhiHAU8 virion has a polyhedral head and a flexible tail and has a small genome (ca. 42.5 kb) with double-stranded DNA and cohesive ends. PhiHAU8 was most stable at 4 degrees C in Difco nutrient broth within a pH range of 6 to 12. PhiHAU8 plaque formation on Micromonospora sp. strain 40027 was optimal with 32 mM Ca(2+) and 30 mM Mg(2+). A lysogen, LXH8, was isolated from turbid plaques, and a phasmid derivative that functions as a lambda cosmid vector in Escherichia coli and as a phage in Micromonospora sp. strain 40027 was constructed. Pulsed-field gel electrophoresis of AseI-digested total DNA showed that PhiHAU8 DNA integrates into the 500-kb AseI fragment of Micromonospora sp. strain 40027.
分离出了PhiHAU8，一种温和型小单孢菌噬菌体，它能够感染小单孢菌属菌株40027和A-M-01。PhiHAU8病毒粒子有一个多面体头部和一条灵活的尾部，并且有一个小基因组（约42.5 kb），含双链DNA和粘性末端。PhiHAU8在Difco营养肉汤中，于4℃、pH值6至12的范围内最为稳定。在小单孢菌属菌株40027上形成噬菌斑的最佳条件是含有32 mM Ca(2+)和30 mM Mg(2+)。从浑浊噬菌斑中分离出了一个溶原菌LXH8，并构建了一种在大肠杆菌中作为λ粘粒载体发挥作用、在小单孢菌属菌株40027中作为噬菌体发挥作用的噬菌粒衍生物。用AseI消化总DNA后的脉冲场凝胶电泳显示，PhiHAU8 DNA整合到了小单孢菌属菌株40027的500-kb AseI片段中。
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