Isolation and characterization of a filamentous viruslike particle from Clostridium acetobutylicum NCIB 6444.
A single-stranded 6.6-kb DNA molecule complexed with protein was recovered from the supernatant of Clostridium acetobutylicum NCIB 6444. Electron microscopic examination of the DNA-protein complex revealed the presence of a filamentous viruslike particle, which was designated CAK1. The possible double-stranded plasmidlike replicative form and the single-stranded prophage were also recovered from the cell culture following alkaline lysis. CAK1 was released from the C. acetobutylicum cell culture in the absence of cell lysis. Polyethylene glycol-NaCl coprecipitation of the DNA-protein complex revealed the presence of single-stranded DNA complexed with protein in a manner rendering the DNA resistant to Bal 31 exonuclease. Proteinase treatment of CsCl density gradient-purified CAK1 resulted in recovery of DNase-sensitive single-stranded DNA. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis of CAK1 demonstrated the presence of a 5-kDa major coat protein. Hybridization data indicated that the single-stranded DNA from CAK1 has homology with the M13 phage of Escherichia coli. An examination of various physical properties of CAK1 suggests that it is similar to the filamentous phage recovered from gram-negative microorganisms. Although infectivity or inducibility of CAK1 could not be demonstrated, to our knowledge this represents the first report of a nonlytic filamentous viruslike particle containing single-stranded DNA being recovered from a gram-positive bacterium.
Kim
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AY
Department of Food Science, University of Illinois, Urbana 61801.
Blaschek
H P
HP
D001435:Bacteriophages / Q000648:ultrastructure*; D003013:Clostridium / Q000648:ultrastructure*; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification* / Q000648:ultrastructure; D004277:DNA, Single-Stranded / Q000235:genetics / Q000302:isolation & purification* / Q000648:ultrastructure; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification* / Q000648:ultrastructure; D004926:Escherichia coli / Q000648:ultrastructure; D008854:Microscopy, Electron; D015183:Restriction Mapping; D014780:Viruses / Q000302:isolation & purification / Q000648:ultrastructure*
D004269:DNA, Bacterial; D004277:DNA, Single-Stranded; D004279:DNA, Viral