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J Virol. 1974 September; 14(3): 620-630
Copyright © 1974 American Society for Microbiology. All Rights Reserved.

Double-Length, Circular, Single-Stranded DNA from Filamentous Phage

Frances C. Wheeler, Rolf H. Benzinger and Hermann Bujard

Department of Biology, University of Virginia, Charlottesville, Virginia 22901
Institut fuer Genetik, Universitaet Heidelberg, Germany

ABSTRACT

Wild-type and gene 3 mutant filamentous phage stocks, containing different relative amounts of multiple-length particles, were treated exhaustively with DNase and then were highly purified. The phage DNA was extracted and examined by electron microscopy. In all cases, about 0.03% of the molecules were circular dimers. 3H-labeled phage DNA was separated as to size by sedimentation in a preformed CsCl density gradient. Individual fractions were then examined in the electron microscope, and the percentage of linear and circular monomer and dimer DNAs was determined. A peak of double-length, circular molecules (with the expected sedimentation constant of 38S) was found ahead of the 24S monomer peak. The double-length molecules had been purified 65-fold. As previously found for single-stranded DNA, the contour length of these molecules was strongly dependent upon ionic strength. Possible artifacts were ruled out, and it was shown that the double-length molecules arose from phage particles.

J Virol. 1974 September; 14(3): 620-630
Copyright © 1974 American Society for Microbiology. All Rights Reserved.



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