1887

Abstract

SUMMARY

Individual cloned dIII fragments of vaccinia virus DNA were introduced into cells by DNA-mediated gene transfer. The presence of individual fragments in the different transformants was confirmed by Southern blot hybridization analysis. Several of the transformants were found to express viral sequences at various levels. The sizes of the transcripts containing vaccinia virus sequences were highly heterogeneous, with no discrete species of RNA. Positive clones contained vaccinia virus sequences in both the poly(A) and poly(A) RNA fractions, although the prevalence of these sequences was variable in the two fractions. The S1 nuclease map of the 5′ end of the transcripts from transformants containing the dIII-J fragment revealed a unique 5′ end, similar to RNA from virus-infected cells. In contrast, analysis of the 3′ end of RNAs from these transformants showed a high degree of heterogeneity, which might explain the heterogeneity found in Northern blot patterns. In this report, it is shown for the first time that in cells transformed with vaccinia virus DNA there is proper initiation for, at least, the viral thymidine kinase gene.

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1984-07-01
2024-03-28
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References

  1. Bajszar G., Witter R., Weir J. P., Moss B. 1983; Vaccinia virus thymidine kinase and neighboring genes: mRNAs and polypeptides of wild-type virus and putative nonsense mutants. Journal of Virology 45:62–72
    [Google Scholar]
  2. Baroudy B. M., Venkatesan S., Moss B. 1982; Incompletely base-paired flip-flop terminal loops link the two DNA strands of the vaccinia virus genome into one uninterrupted polynucleotide chain. Cell 28:315–324
    [Google Scholar]
  3. Berk A. J., Sharp P. A. 1977; Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of SI endo nuclease-digested hybrids. Cell 12:721–732
    [Google Scholar]
  4. Birnboim H. C., Doly J. 1979; A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Research 7:1513–1523
    [Google Scholar]
  5. Cabrera C. V., Esteban M., Mccarron R., Mcallister W. T., Holowczak J. A. 1978; Vaccinia virus transcription: hybridization of mRNA to restriction fragments of vaccinia DNA. Virology 86:102–114
    [Google Scholar]
  6. Colbere-Garapin F., Horodniceanu F., Kourilsky P., Garapin A. C. 1981; A new dominant hybrid selective marker for higher eukaryotic cells. Journal of Molecular Biology 150:1–14
    [Google Scholar]
  7. Corces V., Pellicer A., Axel R., Meselson M. 1981; Integration, transcription, and control of a Drosophila heat shock gene in mouse cells. Proceedings of the National Academy of Sciences, U,. S,. A 78:7038–7042
    [Google Scholar]
  8. Dales S., Pogo B. G. T. 1981 Biology of Poxviruses 109 pp. Virology Monographs vol 18: Edited by Kingsbury D. W., zur Hausen H. Wien & New York: Springer-Verlag;
    [Google Scholar]
  9. Graham F. L., Vander Eb A. J. 1973; A new technique for theassay of infectivity of human adenovirus 5 DNA. Virology 52:456–461
    [Google Scholar]
  10. Grunstein M., Hogness D. S. 1975; Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proceedings of the National Academy of Sciences, U,. S,. A 72:3961–3965
    [Google Scholar]
  11. Hruby D., Ball L. A. 1982; Mapping and identification of the vaccinia virus thymidine kinase gene. Journal of Virology 43:403–409
    [Google Scholar]
  12. Hruby D. E., Maki R. A., Miller D., Ball L. A. 1983; Fine structure analysis and nucleotide sequence of the vaccinia virus thymidine kinase gene. Proceedings of the National Academy of Sciences, U,. S,. A 80:3411–3415
    [Google Scholar]
  13. Littlefield J. W. 1965; Studies on thymidine kinase in cultured fibroblasts. Biochimicia et biophystca acta 95:14–22
    [Google Scholar]
  14. Lowy I., Pellicer A., Jackson J. F., Sim G. K., Silverstein S., Axel R. 1980; Isolation of transforming DNA: cloning the hamster aprt gene. Cell 22:817–824
    [Google Scholar]
  15. Maitland N. J., Mcdougall J. K. 1977; Biochemical transformation of mouse cells by fragments of herpes simplex virus DNA. Cell 11:233–241
    [Google Scholar]
  16. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  17. Moss B. 1974; Reproduction of the poxviruses. In Comprehensive Virology vol 3: pp 405–474 Edited by Fraenkel-Conrat H., Wagner R. R. New York: Plenum Press;
    [Google Scholar]
  18. Pellicer A., Esteban M. 1982; Gene-transfer, stability and biochemical properties of animal cells transformed with vaccinia DNA. Virology 122:363–380
    [Google Scholar]
  19. Pellicer A., Wigler M., Axel R., Silverstein S. 1978; The transfer and stable integration of the HSV thymidine kinase gene into mouse cells. Cell 14:133–141
    [Google Scholar]
  20. Pickup D. J., Bastia D., Joklik W. K. 1983; Cloning of the terminal loop of vaccinia virus DNA. Virology 124:215–217
    [Google Scholar]
  21. Southern E. M. 1975; Detection of specific sequences among DNA fragments separated by gel electrophoresis. Journal of Molecular Biology 98:503–517
    [Google Scholar]
  22. Thomas P. S. 1980; Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proceedings of the National Academy of Sciences, U,. S,. A 77:5201–5205
    [Google Scholar]
  23. Twigg A. J., Sherratt D. 1980; Trans-complementable copy-number mutants of plasmid ColEl. Nature, London 283:216–218
    [Google Scholar]
  24. Venkatesan S., Baroudy B. M., Moss B. 1981; Distinctive nucleotide sequences adjacent to multiple initiation and termination sites of an early vaccinia virus gene. Cell 125:805–813
    [Google Scholar]
  25. Venkatesan S., Gershowitz A., Moss B. 1982; Complete nucleotide sequences of two adjacent early vaccinia virus genes located within the inverted terminal repetition. Journal of Virology 44:637–646
    [Google Scholar]
  26. Weir I. P., Moss R. 1983; Nucleotide sequence of the vaccinia virus thymidine kinase gene and the nature of spontaneous frameshift mutations. Journal of Virology 46:530–537
    [Google Scholar]
  27. Weir J. P., Bajszar G., Moss B. 1982; Mapping of the vaccinia virus thymidine kinase gene by marker rescue and by cell-free translation. Proceedings of the National Academy of Sciences, U,. S,. A 79:1210–1214
    [Google Scholar]
  28. Weiss R. A., Teich N. M., Varmus H., Coffin J. M. (editors) 1982 Molecular Biology of Tumor Viruses, Part 3: RNA Tumor Viruses 2nd edn New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  29. Wigler M., Silverstein S., Lee L. S., Pellicer A., Cheng Y-C., Axel R. 1977; Transfer of purified herpes simplex virus thymidine kinase gene to cultured mouse cells. Cell 11:223–232
    [Google Scholar]
  30. Wigler M., Pellicer A., Silverstein S., Axel R., Ulaub G., Chasin L. 1979a; DNA-mediated transfer of the adenine phosphoribosyl-transferase locus into mammalian cells. Proceedings of the National Academy of Sciences, U,. S,. A 76:1373–1376
    [Google Scholar]
  31. Wigler M., Sweet R., Sim G. K., Wold B., Pellicer A., Lacy E., Maniatis T., Silverstein S., Axel R. 1979b; Transformation of mammalian cells with genes from procaryotes and eucaryotes. Cell 16:777–785
    [Google Scholar]
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