Summary
Complementation experiments were performed with ss and RF DNA1 of bacteriophage ϕX174. Spheroplasts of Escherichia coli K12W6 were mixedly infected under restrictive conditions with
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a)
RF DNA's of mutants in different cistrons,
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b)
ss DNA's of mutants in different cistrons,
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c)
RF DNA of a mutant in one cistron and ss DNA of a mutant in another cistron.
In case a) complementation could be demonstrated unambiguously. The phage yield after complementation amounts to about 1–5 percent of the yield of each mutant under permissive conditions. In case b) the ratio of the total yield over the yield of the selfed controls was the same as in case a) but amounts to only 0.5 percent of the yield under permissive conditions. Moreover, a rather high percentage of wild type particles appeared in the burst. Therefore we could not distinguish between rescue of mutant DNA and complementation. Case c) was intermediate between a) and b). These complementation patterns are discussed.
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References
Borrias, W. E., Pol, J. H. van de, Vate, C. van de, Arkel, G. A. van: Complementation experiments between conditional lethal mutants of bacteriophage ϕX 174. Molec. gen. Genet. 105, 152–163 (1969).
Goulian, M., Kornberg, A.: Enzymatic synthesis of DNA, XXIII. Synthesis of circular replicative form of phage ϕX174 DNA. Proc. nat. Acad. Sci. (Wash.) 58, 1723–1730 (1967).
— and Sinsheimer, R. L.: Enzymatic synthesis of DNA, XXIV. Synthesis of infectious phage ϕX174 DNA. Proc. nat. Acad. Sci. (Wash.) 58, 2321–2328 (1967).
Guthrie, G. D., Sinsheimer, R. L.: Infection of protoplasts of Escherichia coli by subviral particles of bacteriophage ϕX174. J. molec. Biol. 2, 297–305 (1960).
—: Observations on the infection of bacterial protoplasts with deoxyribonucleic acid bacteriophage ϕX174. Biochem. biophys. Acta (Amst.) 72, 290–297 (1963).
Hayashi, M., Hayashi, M. N., Spiegelman, S.: Restriction of in vivo genetic transcription to one of the complementary strands of DNA. Proc. nat. Acad. Sci. (Wash.) 50, 664–672 (1963).
Hayashi, Y., Hayashi, M.: Fractionation of ϕX174 specific messenger RNA. Cold Spr. Harb. Symp. quant. Biol. 35, 171–177 (1970).
Hutchison, C. A., III: Bacteriophage ϕX174: viral genes and functions. Thesis, California Institute of Technology, Pasadena (1969).
Jansz, H. S., Pouwels, P. H., Schiphorst, J.: Preparation of double stranded DNA (replicative form) of bacteriophage ϕX174: a simplified method. Biochim. biophys. Acta (Amst.) 123, 626–627 (1966b).
—, Rotterdam, C. van, Cohen, J. A.: Genetic recombination of bacteriophage ϕX174 in spheroplasts. Biochim. biophys. Acta (Amst.) 119, 276–288 (1966a).
Knippers, R.: DNA polymerase II. Nature (Lond.) 228, 1050–1053 (1970).
—, Razin, A., Davis, R.: The process of infection with bacteriophage ϕX174, XXIX. In vivo studies on the synthesis of the single stranded DNA of progeny ϕX174 bacteriophage. J. molec. Biol. 45, 237–263 (1969).
—, Sinsheimer, R. L.: The process of infection with bacteriophage ϕX174, XX. Attachment of the parental DNA of bacteriophage ϕX174 to a fast sedimenting cell component. J. molec. Biol. 34, 17–29 (1968a).
—: The process of infection with bacteriophage ϕX174, XXIII. DNA synthesis in cells infected by a coat protein mutant. J. molec. Biol. 35, 591–605 (1968b).
—, Strätling, W.: The DNA replicating capacity of isolated Escherichia coli cell wall-membrane complexes. Nature (Lond.) 226, 713–717 (1970).
Lindqvist, B. H., Sinsheimer, R. L.: The process of infection with bacteriophage ϕX174, XVI. Synthesis of the replicative form and its relationship to viral single stranded DNA synthesis. J. molec. Biol. 32, 285–302 (1968).
Mitra, S., Reichard, P., Inman, R. B., Bertsch, L. L., Kornberg, A.: Enzymatic synthesis of DNA, XXII. Replication of a circular single-stranded DNA template by DNA polymerase of Escherichia coli. J. molec. Biol. 24, 429–447 (1967).
Salivar, W. O., Sinsheimer, R. L.: Intracellular location and number of replicating parental DNA molecules of bacteriophages λ and ϕX174 J. molec. Biol. 41, 39–65 (1969).
Schleser R., Tessman, E. S., Casaday, G.: Protein synthesis by a mutant of phage S13 blocked in DNA synthesis. Virology 38, 166–173 (1969).
Sinsheimer, R. L.: Bacteriophage ϕX174 and related viruses. Progr. Nucl. Acid Res. and molec. Biol. 8, 115–169 (1968).
Weisbeek, P. J., Pol, J. H. van de: Biological activity of ϕX174 replicative form DNA fragments. Biochim. biophys. Acta (Amst.) 224, 328–338 (1970).
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Borrias, W.E., van de Pol, J.H. Genetic complementation between bacteriophage ϕX174 DNA molecules in spheroplasts. Molec. Gen. Genet. 114, 59–67 (1972). https://doi.org/10.1007/BF00268747
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DOI: https://doi.org/10.1007/BF00268747