1887

Abstract

A cDNA clone of the tobacco mosaic virus 30K movement protein (MP) gene was constructed and introduced into an nuclear polyhedrosis baculovirus expression vector. Infection of cells with the vector resulted in the synthesis of low levels of MP, which was detected by anti-MP serum as two closely related species of approximately 34K and a third species of 32K. The authenticity of the recombinant MP was confirmed by comparison of the protein, on the basis of migration during SDS-PAGE, with authentic MP from several sources. It appeared that the recombinant MP was not modified by -linked glycosylation, but was phosphorylated. The recombinant MP was produced in both a phosphorylated and an unphosphorylated state and the former species was shown to comigrate with plant-expressed MP during SDS-PAGE.

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1991-11-01
2021-10-28
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References

  1. Atkins D., Hull R., Wells B., Roberts K., Moore P., Beachy R. N. 1991; The tobacco mosaic virus 30K movement protein in transgenic tobacco plants is localized to plasmodesmata. Journal of General Virology 72:209–211
    [Google Scholar]
  2. Brown M., Faulkner P. 1977; A plaque assay for nuclear polyhedrosis viruses using a solid overlay. Journal of General Virology 36:361–364
    [Google Scholar]
  3. Citovsky V., Knorr D., Schuster G., Zambryski P. 1990; The P30 movement protein of tobacco mosaic virus is a single-stranded nucleic acid binding protein. Cell 60:637–647
    [Google Scholar]
  4. Deom C. M., Oliver M. J., Beachy R. N. 1987; The 30-kilodalton gene product of tobacco mosaic virus potentiates virus movement. Science 237:389–394
    [Google Scholar]
  5. Deom C. M., Schubert K. R., Wolf S., Holt C. A., Lucas W. J., Beachy R. N. 1990; Molecular characterization and biological function of the movement protein of tobacco mosaic virus in transgenic plants. Proceedings of the National Academy of Sciences, U.S.A. 87:3284–3288
    [Google Scholar]
  6. Dietz K.-J., Kaiser G., Martinoia E. 1988; Characterization of vacuolar polypeptides of barley mesophyll cells by two-dimensional gel electrophoresis and by their affinity to lectins. Planta 176:362–367
    [Google Scholar]
  7. Dietzschold B., Cox J. H., Schneider L. G. 1979; Rabies virus strains: a comparison study by polypeptide analysis of vaccine strains with different pathogenic patterns. Virology 98:63–75
    [Google Scholar]
  8. Goelet P., Lomonossoff G. P., Butler P. J. G., Akam M. E., Gait M. J., Karn J. 1982; Nucleotide sequence of tobacco mosaic virus RNA. Proceedings of the National Academy of Sciences, U.S.A. 79:5818–5822
    [Google Scholar]
  9. Hirt B. 1967; Selective extraction of polyoma DNA from infected mouse cell cultures. Journal of Molecular Biology 26:365–369
    [Google Scholar]
  10. Hull R. 1989; The movement of viruses in plants. Annual Review of Phytopathology 27:213–240
    [Google Scholar]
  11. Kunkel T. A. 1985; Rapid and efficient site-specific mutagenesis without phenotypic selection. Proceedings of the National Academy of Sciences, U.S.A. 82:488–192
    [Google Scholar]
  12. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  13. Luckow V. A., Summers M. D. 1988; Trends in the development of baculovirus expression vectors. Bio/Technology 6:47–55
    [Google Scholar]
  14. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  15. Matsuura Y., Possee R. D., Overton H. A., Bishop D. H. L. 1987; Baculovirus expression vectors: the requirements for high level expression of proteins, including glycoproteins. Journal of General Virology 68:1233–1250
    [Google Scholar]
  16. Matsuura Y., Miyamoto M., Sato T., Morita C., Yasui K. 1989; Characterization of Japanese encephalitis envelope protein expressed by recombinant baculoviruses. Virology 173:674–682
    [Google Scholar]
  17. Miller L. K. 1988; Baculoviruses as gene expression vectors. Annual Review of Microbiology 42:177–199
    [Google Scholar]
  18. Moser O., Gagey M. -J., Godefroy-Colburn T., Stussi-Garaud C., Ellwart-Tschurtz M., Nitschko H., Mundry K. -W. 1988; The fate of the transport protein of tobacco mosaic virus in systemic and hypersensitive tobacco hosts. Journal of General Virology 69:1367–1373
    [Google Scholar]
  19. Pelham H. R. B., Jackson R. J. 1976; An efficient mRNA dependent translation system from reticulocyte lysates. European Journal of Biochemistry 67:493–497
    [Google Scholar]
  20. Smith G. E., Summers M. D. 1978; Analysis of baculovirus genomes with restriction endonucleases. Virology 89:517–527
    [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. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences, U.S.A. 76:4350–4354
    [Google Scholar]
  23. Watanabe Y., Emori Y., Ooshika I., Meshi T., Ohno T., Okada Y. 1984; Synthesis of TMV specific RNAs and proteins at the early stage of infection in tobacco protoplasts: transient expression of the 30K protein and its mRNA. Virology 133:18–24
    [Google Scholar]
  24. Wolf S., Deom C. M., Beachy R. N., Lucas W. J. 1989; Movement protein of tobacco mosaic virus modifies plasmodesmatal size exclusion limit. Science 246:377–379
    [Google Scholar]
  25. Zaitlin M., Hull R. 1987; Plant virus-host interactions. Annual Review of Plant Physiology 38:291–315
    [Google Scholar]
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