1887

Abstract

DNA fragments from promoter regions of the geminivirus, African cassava mosaic virus, were cloned into pG1, a vector based on pUC18, producing transcriptional fusions with the β-glucuronidase (GUS) gene and nopaline synthase termination sequence. The activity of each promoter construct was assessed by analysing the transient expression of GUS in protoplasts. The results demonstrated that constructs containing the common region of DNA A showed much stronger promoter activity in the complementary sense than in the viral sense. These results were supported by the analysis of promoter activity in transgenic plants. In comparison, in protoplasts a region upstream of the AC2 open reading frame was shown to have moderate promoter activity. Unlike DNA A, the complementary sense DNA B promoter constructs had weak activity; the viral sense DNA B promoter constructs appeared to be regulated by host factors. The implications of these results for the regulation of early and late genes are discussed.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-72-11-2849
1991-11-01
2024-04-25
Loading full text...

Full text loading...

/deliver/fulltext/jgv/72/11/JV0720112849.html?itemId=/content/journal/jgv/10.1099/0022-1317-72-11-2849&mimeType=html&fmt=ahah

References

  1. Benfey P. N., Ren L., Chua N. -H. 1990; Combinatorial and synergistic properties of CaMV 35S enhancer subdomains. EMBO Journal 9:1685–1696
    [Google Scholar]
  2. Bevan M. 1984; Binary Agrobacterium vectors for plant transformation. Nucleic Acids Reserch 12:8711–8721
    [Google Scholar]
  3. Bradford M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72:248–254
    [Google Scholar]
  4. Davies J. W., Stanley J. 1989; Geminivirus genes and vectors. Trends in Genetics 5:77–81
    [Google Scholar]
  5. Davies J. W., Townsend R., Stanley J. 1987; The structure, expression, functions and possible explanation of geminivirus genomes. In Plant DNA Infectious Agents pp 31–52 Edited by Hohn T., Schell J. Wien & New York: Springer-Verlag;
    [Google Scholar]
  6. Depicker A., Stachel S., Dhaese P., Zambryski P., Goodman H. 1982; Nopaline synthase: transcript mapping and DNA sequence. Journal of Molecular and Applied Genetics 1:561–575
    [Google Scholar]
  7. Etessami P., Callis R., Ellwood S., Stanley J. 1988; Delimitation of essential genes of cassava latent virus DNA 2. Nucleic Acids Research 16:4811–4829
    [Google Scholar]
  8. Fenoll C., Black D. M., Howell S. H. 1988; The intergenic region of maize streak virus contains promoter elements involved in rightward transcription of the viral genome. EMBO Journal 7:1589–1596
    [Google Scholar]
  9. Fenoll C., Schwarz J. J., Black D. M., Schneider M., Howell S. H. 1990; The intergenic region of maize streak virus contains a GC rich element that activates rightward transcription and binds maize nuclear factors. Plant Molecular Biology 15:865–877
    [Google Scholar]
  10. Fillati J.-A., Sellmer J., McCown B., Haissig B., Comai L. 1987; Agrobacterium-mediated transformation and regeneration of Populus. Molecular and General Genetics 206:192–199
    [Google Scholar]
  11. Horsch R. B., Fry J. E., Hoffman N. L., Eichholtz D., Rogers S. G., Fraley R. T. 1985; A simple and general method for transferring genes into plants. Science 227:1229–1231
    [Google Scholar]
  12. Jefferson R. A. 1987; Assaying chimeric genes in plants: the GUS gene fusion system. Plant Molecular Biology Reporter 5:387–405
    [Google Scholar]
  13. Kosugi S., Ohashi Y., Nakajima K., Arai Y. 1990; An improved assay for β-glucuronidase in transformed cells: methanol almost completely suppresses a putative endogenous β-glucuronidase activity. Plant Science 70:133–140
    [Google Scholar]
  14. Morris B. A. M., Richardson K. A., Andersen M. T., Gardner R. C. 1988; Cassava latent virus infections mediated by the Ti plasmid of Agrobacterium tumefaciens containing either monomeric or dimeric viral DNA. Plant Molecular Biology 11:795–803
    [Google Scholar]
  15. Morris B., Richardson K., Eddy P., Zhan X., Haley A., Gardner R. 1991; Mutagenesis of the AC3 open reading frame of African cassava mosaic virus DNA A reduces DNA B replication and ameliorates disease symptoms. Journal of General Virology 72:1205–1213
    [Google Scholar]
  16. Rabussay D., Uher L., Bates G., Piastuch W. 1987; Electroporation of mammalian and plant cells. Focus 9:1–5
    [Google Scholar]
  17. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: A laboratory Manual, 2nd. edn New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  18. Stanley J. 1985; The molecular biology of geminiviruses. Advances in Virus Research 30:139–177
    [Google Scholar]
  19. Stanley J., Gay M. R. 1983; Nucleotide sequence of cassava latent virus DNA. Nature, London 301:260–262
    [Google Scholar]
  20. Sunter G., Bisaro D. M. 1991; Transactivation in a geminivirus: AL2 gene product is needed for coat protein expression. Virology 180:416–419
    [Google Scholar]
  21. Townsend R., Stanley J., Curson S. J., Short M. N. 1985; Major polyadenylated transcripts of cassava latent virus and location of the gene encoding coat protein. EMBO Journal 4:33–37
    [Google Scholar]
  22. Ward A., Etessami P., Stanley J. 1988; Expression of a bacterial gene in plants mediated by infectious geminivirus DNA. EMBO Journal 7:1583–1587
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-72-11-2849
Loading
/content/journal/jgv/10.1099/0022-1317-72-11-2849
Loading

Data & Media loading...

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