Transfection of Spheroplasts: Infectious Lambda Prophage DNA Free

Abstract

SUMMARY

High mol. wt. DNA was extracted from lambda lysogens and was shown to be infectious. Its infectivity was due to prophage DNA integrated into the host chromosome rather than to DNA released from mature phage particles, as established by the following criteria: the titre of infectious DNA exceeded by 100-fold the titre of infectious units present before DNA extraction; mild shear selectively reduced prophage DNA infectivity to 2% of the unsheared DNA while lambda phage DNA infectivity retained 50% of its infectivity; DNA extracted from an (lambda c857 ts) lysogen yielded 200 times as many plaques on than on spheroplasts. Thus lambda prophage DNA infectivity depends on expression of the excision gene while the infectivity of non-integrated forms of lambda does not. About 10 genome equivalents of DNA yielded one infectious centre unit in this assay system; this high infectivity should make prophage DNA a useful marker in genetic transformation experiments.

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1978-06-01
2024-03-28
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References

  1. Armentrout R. W., Rutberg L. 1970; Mapping of prophage and mature deoxyribonucleic acid from temperate Bacillus bacteriophage 1/H05 by marker rescue. Journal of Virology 6:760–767
    [Google Scholar]
  2. Armentrout R. W., Rutberg L. 1971; Heat induction of prophage ^105 in Bacillus subtilis: replication of the bacterial and bacteriophage genomes. Journal of Virology 8:455–468
    [Google Scholar]
  3. Barnhart B. J., Cox S. H. 1973; DNA replication of induced prophage in Hemophilus influenzae. Journal of Virology 12:165–176
    [Google Scholar]
  4. Boling M. E., Setlow J. K., Allison D. P. 1972; Bacteriophage of Haemophilus influenzae. I. Differences between infection by whole phage, extracted phage DNA and prophage DNA extracted from lysogenic cells. Journal of Molecular Biology 63:335–348
    [Google Scholar]
  5. Cosloy S. D., Oishi M. 1973; Genetic transformation in Escherichia coli K12. Proceedings of the National Academy of Sciences of the United States of America 70:84–87
    [Google Scholar]
  6. Gottesman S., Gottesman M. E. 1975a; Elements involved in site-specific recombination in bacteriophage lambda. Journal of Molecular Biology 91:489–499
    [Google Scholar]
  7. Gottesman S., Gottesman M. 1975b; Excision of prophage lambda in a cell-free system. Proceedings of the National Academy of Sciences of the United States of America 72:2188–2192
    [Google Scholar]
  8. Harm W., Rupert C. S. 1963; Infection of transformable cells of Haemophilus influenzae by bacteriophage and bacteriophage DNA. Zeitschrift für Vererbungslehre 94:336–348
    [Google Scholar]
  9. Henner W. D., Kleber I., Benzinger R. 1973; Transfection of E. coli spheroplasts. III. Facilitation of transfection and stabilization of spheroplasts by different basic polymers. Journal of Virology 12:741–747
    [Google Scholar]
  10. Nash H. A. 1974; Aatt B-att P, a lambda derivative containing both sites involved in integrative recombination. Virology 57:207–740
    [Google Scholar]
  11. Nash H. A. 1975a; Integrative recombination in bacteriophage lambda: analysis of recombinant DNA. Journal of Molecular Biology 91:501–514
    [Google Scholar]
  12. Nash H. A. 1975b; Integrative recombination of bacteriophage lambda in vitro. Proceedings of the National Academy of Sciences of the United States of America 72:1072–1076
    [Google Scholar]
  13. Peterson A., Rutberg L. 1969; Linked transformation of bacterial and prophage markers in Bacillus subtilis 168 lysogenic for bacteriophage ϕ 105. Journal of Bacteriology 98:874–876
    [Google Scholar]
  14. Quinn W. G., Sueoka N. 1970; Symmetric replication of the B. subtilis chromosome. Proceedings of the National Academy of Sciences of the United States of America 67:717–723
    [Google Scholar]
  15. Romig W. R. 1968; Infectivity of Bacillus subtilis bacteriophage deoxyribonucleic acids from mature particles and lysogenic hosts. Bacteriological Reviews 32:349–357
    [Google Scholar]
  16. Rutberg L. 1969; Mapping of a temperature bacteriophage active on Bacillus subtilis. Journal of Virology 3:38–44
    [Google Scholar]
  17. Rutberg L. 1971; Heat induction of prophage ϕ 105 in Bacillus subtilis: bacteriophage-induced bidirectional replication of the bacterial chromosome. Journal of Virology 12:9–12
    [Google Scholar]
  18. Rutberg L. 1973; Heat induction of prophage 56105 in Bacillus subtilis: bacteriophage-induced bidirectional replication of the bacterial chromosome. Journal of Virology 12:9–12
    [Google Scholar]
  19. Rutberg L., Hoch J. A., Spizizen J. 1969; Mechanism of transfection with deoxyribonucleic acid from the temperate Bacillus bacteriophage 56105. Journal of Virology 4:50–57
    [Google Scholar]
  20. Schekman R. W., Iwaya M., Bromstrup K., Denhardt D. T. 1971; The mechanism of replication of ϕ X 174 single-stranded DNA III. An enzymic study of the structure of the replicative form II DNA. Journal of Molecular Biology 57:177–199
    [Google Scholar]
  21. Shimada K., Weisberg R. A., Gottesman M. E. 1972; Prophage lambda at unusual locations. I. Location of the secondary attachment sites and the properties of the lysogens. Journal of Molecular Biology 63482–502
    [Google Scholar]
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