Tilley B C, Meyertons J L, Lechevalier M P
Waksman Institute of Microbiology, Rutgers University, Piscataway, NJ 08855-0759.
J Ind Microbiol. 1990 Apr-May;5(2-3):167-82. doi: 10.1007/BF01573867.
A temperate actinophage was isolated from soil using the gentamicin-producing microorganism, Micromonospora purpurea ATCC 15835 as host. The characterization of the phage represents the initial step in its development as a cloning vector. The phage isolated, MPphiWR-1, formed red- to purple-pigmented turbid plaques. Cells isolated from these plaques were resistant to superinfection with lytic mutants of MPphiWR-1. Southern blots of genomic DNA from a resistant culture showed that MPphiWR-1 integrated into the host genome. The phage was UV- or Mitomycin C-inducible. The integration, resistance to superinfection and inducibility indicated a lysogenic relationship with the host. Using MPphiE-RCPM, a lytic derivative, the phage host range was demonstrated to include members of three genera: one species each of Ampullariella and Catellatospora, and 12 species of Micromonospora. The phage belonged to Ackerman's B1 morphotype having an isometric head and a flexible noncontractile tail. The density of the phage was 1.525 g/cc. Restriction site mapping demonstrated that the phage DNA was 57.9 kb long and had cohesive ends. Using EDTA enrichment, viable mutants with deletions of at least 3.5 kb were isolated and mapped. Phage adsorption, sensitivities and plating efficiency were investigated. Non-liposome PEG-mediated transfection was demonstrated.
利用产生庆大霉素的微生物——紫色小单孢菌ATCC 15835作为宿主，从土壤中分离出一种温和性肌尾噬菌体。对该噬菌体的特性描述是将其开发为克隆载体的第一步。分离得到的噬菌体MPphiWR-1形成了红色至紫色的浑浊噬菌斑。从这些噬菌斑中分离出的细胞对MPphiWR-1的裂解突变体的超感染具有抗性。来自抗性培养物的基因组DNA的Southern印迹显示MPphiWR-1整合到宿主基因组中。该噬菌体可被紫外线或丝裂霉素C诱导。整合、对超感染的抗性和可诱导性表明其与宿主存在溶原性关系。使用裂解衍生物MPphiE-RCPM，证明该噬菌体的宿主范围包括三个属的成员：壶孢菌属和链孢囊菌属各一个种，以及小单孢菌属的12个种。该噬菌体属于Ackerman的B1形态型，具有等轴头和灵活的非收缩尾。噬菌体的密度为1.525 g/cc。限制性酶切位点图谱显示噬菌体DNA长57.9 kb，具有粘性末端。使用EDTA富集法，分离并定位了至少缺失3.5 kb的存活突变体。研究了噬菌体的吸附、敏感性和平板效率。证明了非脂质体聚乙二醇介导的转染。
