1887

Abstract

A new gene-expression system based on RNA-5 of (BNYVV) was constructed to allow the expression of recombinant proteins in virally infected cells. Replication and expression levels of the RNA-5-based replicon containing the green fluorescence protein (GFP) gene were compared with those obtained with the well-characterized RNA-3-derived replicon (Rep-3). When RNA-3 and/or RNA-4 BNYVV RNAs were added to the inoculum, the expression levels of RNA-5-encoded GFP were considerably reduced. To a lesser extent, RNA-3-derived GFP expression was also affected by the presence of RNA-4 and -5. Both RNA-3- and RNA-5-derived molecules were able to express proteins within the same infected cells. Together with Rep-3, the RNA-5-derived replicon thus provides a new tool for the co-expression of different recombinant proteins. In , Rep-5-GFP was able to move in systemic tissues in the presence of RNA-3 and thus provides a new expression system that is not restricted to the inoculated leaves.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.80720-0
2005-02-01
2019-11-13
Loading full text...

Full text loading...

/deliver/fulltext/jgv/86/2/vir860463.html?itemId=/content/journal/jgv/10.1099/vir.0.80720-0&mimeType=html&fmt=ahah

References

  1. Awram, P., Gardner, R. C., Forster, R. L. & Bellamy, A. R. ( 2002; ). The potential of plant viral vectors and transgenic plants for subunit vaccine production. Adv Virus Res 58, 81–124.
    [Google Scholar]
  2. Bleykasten-Grosshans, C., Guilley, H., Bouzoubaa, S., Richards, K. E. & Jonard, G. ( 1997; ). Independent expression of the first two triple gene block proteins of beet necrotic yellow vein virus complements virus defective in the corresponding gene but expression of the third protein inhibits viral cell-to-cell movement. Mol Plant Microbe Interact 10, 240–246.[CrossRef]
    [Google Scholar]
  3. Erhardt, M., Morant, M., Ritzenthaler, C., Stussi-Garaud, C., Guilley, H., Richards, K., Jonard, G., Bouzoubaa, S. & Gilmer, D. ( 2000; ). P42 movement protein of Beet necrotic yellow vein virus is targeted by the movement proteins P13 and P15 to punctate bodies associated with plasmodesmata. Mol Plant Microbe Interact 13, 520–528.[CrossRef]
    [Google Scholar]
  4. Gilmer, D., Richards, K., Jonard, G. & Guilley, H. ( 1992; ). cis-active sequences near the 5′-termini of beet necrotic yellow vein virus RNAs 3 and 4. Virology 190, 55–67.[CrossRef]
    [Google Scholar]
  5. Gleba, Y., Marillonnet, S. & Klimyuk, V. ( 2004; ). Engineering viral expression vectors for plants: the ‘full virus' and the ‘deconstructed virus' strategies. Curr Opin Plant Biol 7, 182–188.[CrossRef]
    [Google Scholar]
  6. Hehn, A., Bouzoubaa, S., Bate, N., Twell, D., Marbach, J., Richards, K., Guilley, H. & Jonard, G. ( 1995; ). The small cysteine-rich protein P14 of beet necrotic yellow vein virus regulates accumulation of RNA 2 in cis and coat protein in trans. Virology 210, 73–81.[CrossRef]
    [Google Scholar]
  7. Lauber, E., Bleykasten-Grosshans, C., Erhardt, M., Bouzoubaa, S., Jonard, G., Richards, K. E. & Guilley, H. ( 1998a; ). Cell-to-cell movement of beet necrotic yellow vein virus: I. Heterologous complementation experiments provide evidence for specific interactions among the triple gene block proteins. Mol Plant Microbe Interact 11, 618–625.[CrossRef]
    [Google Scholar]
  8. Lauber, E., Guilley, H., Tamada, T., Richards, K. E. & Jonard, G. ( 1998b; ). Vascular movement of beet necrotic yellow vein virus in Beta macrocarpa is probably dependent on an RNA 3 sequence domain rather than a gene product. J Gen Virol 79, 385–393.
    [Google Scholar]
  9. Lauber, E., Jonard, G., Guilley, H. & Gilmer, D. ( 1999; ). Effects of structural modifications upon the accumulation in planta of replicons derived from beet necrotic yellow vein virus RNA 3. Arch Virol 144, 1201–1208.[CrossRef]
    [Google Scholar]
  10. Lemaire, O., Merdinoglu, D., Valentin, P., Putz, C., Ziegler-Graff, V., Guilley, H., Jonard, G. & Richards, K. ( 1988; ). Effect of beet necrotic yellow vein virus RNA composition on transmission by Polymyxa betae. Virology 162, 232–235.[CrossRef]
    [Google Scholar]
  11. Niesbach-Klösgen, U., Guilley, H., Jonard, G. & Richards, K. ( 1990; ). Immunodetection in vivo of beet necrotic yellow vein virus-encoded proteins. Virology 178, 52–61.[CrossRef]
    [Google Scholar]
  12. Pogue, G. P., Lindbo, J. A., Dawson, W. O. & Turpen, T. H. ( 1998; ). Tobamovirus transient expression vectors: tools for plant biology and high-level expression of foreign protein in plants. In Plant Molecular Biology Manual, 2nd edn, pp. 1–27. Edited by S. B. Gelvin & R. A. Schilperoort. Dordrecht: Kluwer.
  13. Pogue, G. P., Lindbo, J. A., Garger, S. J. & Fitzmaurice, W. P. ( 2002; ). Making an ally from an enemy: plant virology and the new agriculture. Annu Rev Phytopathol 40, 45–74.[CrossRef]
    [Google Scholar]
  14. Porta, C. & Lomonossoff, G. P. ( 2002; ). Viruses as vectors for the expression of foreign sequences in plants. Biotechnol Genet Eng Rev 19, 245–291.[CrossRef]
    [Google Scholar]
  15. Quillet, L., Guilley, H., Jonard, G. & Richards, K. ( 1989; ). In vitro synthesis of biologically active beet necrotic yellow vein virus RNA. Virology 172, 293–301.[CrossRef]
    [Google Scholar]
  16. Vetter, G., Hily, J.-M., Klein, E., Schmidlin, L., Haas, M., Merkle, T. & Gilmer, D. ( 2004; ). Nucleo-cytoplasmic shuttling of the beet necrotic yellow vein virus RNA-3-encoded p25 protein. J Gen Virol 85, 2459–2469.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.80720-0
Loading
/content/journal/jgv/10.1099/vir.0.80720-0
Loading

Data & Media loading...

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error