Characterization of a temperate actinophage, MPphiWR-1, capable of infecting Micromonospora purpurea ATCC 15835.
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.
Waksman Institute of Microbiology, Rutgers University, Piscataway, NJ 08855-0759.
B C
BC
Tilley
J L
JL
Meyertons
M P
MP
Lechevalier
D000327:Adsorption; D001435:Bacteriophages / Q000235:genetics / Q000502:physiology* / Q000648:ultrastructure; D015139:Blotting, Southern; D003001:Cloning, Molecular; D003470:Culture Media; D004252:DNA Mutational Analysis; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004279:DNA, Viral / Q000032:analysis* / Q000378:metabolism; D004352:Drug Resistance, Microbial; D005822:Genetic Vectors; D008242:Lysogeny; D008847:Micromonospora; D008854:Microscopy, Electron; D015183:Restriction Mapping; D012988:Soil Microbiology; D013696:Temperature; D014162:Transfection
D003470:Culture Media; D004279:DNA, Viral; D004262:DNA Restriction Enzymes