Induction of multiple prophages from a marine bacterium: a genomic approach.
Approximately 70% of sequenced bacterial genomes contain prophage-like structures, yet little effort has been made to use this information to determine the functions of these elements. The recent genomic sequencing of the marine bacterium Silicibacter sp. strain TM1040 revealed five prophage-like elements in its genome. The genomes of these prophages (named prophages 1 to 5) are approximately 74, 30, 39, 36, and 15 kb long, respectively. To understand the function of these prophages, cultures of TM1040 were treated with mitomycin C to induce the production of viral particles. A significant increase in viral counts and a decrease in bacterial counts when treated with mitomycin C suggested that prophages were induced from TM1040. Transmission electron microscopy revealed one dominant type of siphovirus, while pulsed-field gel electrophoresis demonstrated two major DNA bands, equivalent to 35 and 75 kb, in the lysate. PCR amplification with primer sets specific to each prophage detected the presence of prophages 1, 3, and 4 in the viral lysate, suggesting that these prophages are inducible, but not necessarily to the same level, while prophages 2 and 5 are likely defective or non-mitomycin C-inducible phages. The combination of traditional phage assays and modern microbial genomics provides a quick and efficient way to investigate the functions and inducibility of prophages, particularly for a host harboring multiple prophages with similar sizes and morphological features.
Chen
Feng
F
Center of Marine Biotechnology, University of Maryland Biotechnology Institute, Baltimore, MD 21202, USA. chenf@umbi.umd.edu
Wang
Kui
K
Stewart
Jeneen
J
Belas
Robert
R
D001435:Bacteriophages / Q000235:genetics / Q000502:physiology*; D004279:DNA, Viral / Q000032:analysis / Q000302:isolation & purification; D016680:Genome, Bacterial; D016679:Genome, Viral; D046529:Microscopy, Electron, Transmission; D016685:Mitomycin / Q000494:pharmacology*; D016133:Polymerase Chain Reaction; D039002:Prophages / Q000235:genetics / Q000502:physiology*; D041902:Rhodobacteraceae / Q000254:growth & development / Q000821:virology*; D012623:Seawater / Q000382:microbiology*; D017906:Siphoviridae / Q000235:genetics / Q000502:physiology; D014771:Virion / Q000302:isolation & purification; D014775:Virus Activation / Q000187:drug effects
D004279:DNA, Viral; D016685:Mitomycin