Skip to main content
SpringerLink
Log in

Effects of promoter, intron and enhancer elements on transient gene expression in sugar-cane and carrot protoplasts

  • Short Communication
  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

Various chimaeric promoter regions coupled to the uidA β-glucuronidase gene were evaluated for transient expression strength following electroporation into sugar-cane (monocot) and carrot (dicot) protoplasts. Multiple enhancer elements increased expression in sugar-cane, by up to 400-fold for the artificial Emu promoter relative to the CaMV 35S promoter. The relative expression strengths of promoters varied substantially between the species. Sugar-cane also differed in some respects from previously tested species in the family Poaceae. For example, in sugar-cane the nopaline synthase and CaMV 35S promoters were of equivalent strength, and insertion of Adh1 intron 1 into the 5′ transcribed region decreased expression strength.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

References

  1. Benfey PN, Ren L, Chua N-H: Combinational and synergistic properties of CaMV 35S enhancer subdomains. EMBO J 9: 1685–1696 (1990).

    Google Scholar 

  2. Birch RG, Maretzki A: Transformation of sugar-cane. In: Bajaj YPS (ed) Plant Protoplasts and Genetic Engineering IV. Biotechnology in Agriculture and Forestry Vol. 23. Springer-Verlag, Heidelberg (1993).

    Google Scholar 

  3. Bower R, Birch RG: Competence for gene transfer by electroporation in a sub-population of protoplasts from uniform carrot cell suspension cultures. Plant Cell Rep 9: 386–389 (1990).

    Google Scholar 

  4. Bower R, Birch RG: Transgenic sugar-cane plants via microprojectile bombardment. Plant J 2: 409–416 (1992).

    Google Scholar 

  5. Bradford M: A rapid and sensitive method for the quantification of microgram quantities of protein using the principle of protein-dye binding. Anal Biochem 72: 248–254 (1976).

    Google Scholar 

  6. Callis J, Fromm M, Walbot V: Introns increase gene expression in cultured maize cells. Genes Devel 1: 1183–1200 (1987).

    Google Scholar 

  7. Cao J, Duan X, McElroy D, Wu R: Regeneration of herbicide resistant transgenic rice plants following microprojectile-mediated transformation of suspension culture cells. Plant Cell Rep 11: 586–591 (1992).

    Google Scholar 

  8. Dennis ES, Gerlach WL, Pryor AJ, Bennetzen JL, Inglis A, Llewellyn D, Sachs MM, Ferl RJ, Peacock WJ: Molecular analysis of the alcohol dehydrogenase (Adh1) gene of maize. Nucl Acids Res 12: 3983–4000 (1984).

    Google Scholar 

  9. Ebert PR, Ha SB, An G: Identification of an essential upstream element in the nopaline synthase promoter by stable and transient expression. Proc Natl Acad Sci USA 83: 5372–5376 (1987).

    Google Scholar 

  10. Ecker JR, Davis RW: Inhibition of gene expression in plant cells by expression of antisense RNA. Proc Natl Acad Sci USA 83: 5372–5376 (1986).

    Google Scholar 

  11. Ellis JG, Llewellyn DJ, Walker JC, Dennis ES, Peacock WJ: The OCS element: a 16 base pair palindrome essential for activity of the octopine synthase enhancer. EMBO J 6: 3203–3208 (1987).

    Google Scholar 

  12. Fang R-X, Nagy F, Sivasubramaniam S, Chua N-H: Multiple cis regulatory elements for maximal expression of the cauliflower mosaic virus 35S promoter in transgenic plants. Plant Cell 1: 141–150 (1989).

    Google Scholar 

  13. Franks T, Birch RG: Gene transfer into intact sugar-cane cells using microprojectile bombardment. Aust J Plant Physiol 18: 471–480 (1991).

    Google Scholar 

  14. Fromm ME, Morrish F, Armstrong C, Williams R, Thomas J, Klein TM: Inheritance and expression of chimaeric genes in the progeny of transgenic maize plants. Bio/technology 8: 833–839 (1990).

    Google Scholar 

  15. Fromm M, Taylor LP, Walbot V: Expression of genes transferred to monocot and dicot plant cells by electroporation. Proc Natl Acad Sci USA 82: 5824–5828 (1985).

    Google Scholar 

  16. Hagio T, Blowers AD, Earle ED: Stable transformation of sorghum cell cultures after bombardment with DNA-coated microprojectiles. Plant Cell Rep 10: 260–264 (1991).

    Google Scholar 

  17. Jefferson RA: Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5: 387–405 (1987).

    Google Scholar 

  18. Junker B, Zimney J, Lührs R, Lörz H: Transient expression of chimaeric genes in dividing and non-dividing cereal protoplasts after PEG-induced DNA uptake. Plant Cell Rep 6: 329–332 (1987).

    Google Scholar 

  19. Kay R, Chan A, Daly M, McPherson J: Duplication of 35S promoter sequences creates a strong enhancer for plant genes. Science 236: 1299–1302 (1987).

    Google Scholar 

  20. Kieth B, Chua N-H: Monocot and dicot pre-mRNAs are processed with different efficiencies in transgenic tobacco. EMBO J 5: 2419–2425 (1986).

    Google Scholar 

  21. Last DI, Brettel RIS, Chamberlain DA, Chaudhury AM, Larkin PJ, Marsh EL, Peacock WJ, Dennis ES: pEmu: an improved vector for gene expression in cereal cells. Theor Appl Genet 81: 581–588 (1991).

    Google Scholar 

  22. Leon P, Planckaert F, Walbot V: Transient gene expression in protoplasts of Phaseolus vulgaris isolated from a cell suspension culture. Plant Physiol 95: 968–972 (1991).

    Google Scholar 

  23. Luehrsen KR, Walbot V: Intron enhancement of gene expression and the splicing efficiency of introns in maize cells. Mol Gen Genet 225: 81–93 (1991).

    Google Scholar 

  24. Lütcke HA, Chow KC, Mickel FS, Moss KA, Kern HF, Scheele GA: Selection of AUG initiation codons differs in plants and animals. EMBO J 6: 43–48 (1987).

    Google Scholar 

  25. Maas C, Laufs J, Grant S, Korfhage C, Werr W: The combination of of a novel stimulatory element in the first exon of the maize Shrunken-1 gene with the following intron 1 enhances reporter gene expression up to 1000-fold. Plant Mol Biol 16: 199–207 (1991).

    Google Scholar 

  26. Maniatis T, Fritsch GF, Sambrook J: Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).

    Google Scholar 

  27. McElroy D, Zhang W, Cao J, Wu R: Isolation of an efficient actin promoter for use in rice transformation. Plant Cell 2: 163–171 (1990).

    Google Scholar 

  28. Nagel RJ, Manners JM, Birch RG: Evaluation of an ELISA assay for rapid detection and quantification of neomycin phosphotransferase II in transgenic plants. Plant Mol Biol Rep 10: 263–272 (1992).

    Google Scholar 

  29. Oard JH, Paige D, Dvorak J: Chimeric gene expression using maize intron in cultured cells of breadwheat. Plant Cell Rep 8: 156–160 (1989).

    Google Scholar 

  30. Rathus C, Birch RG: Optimization of conditions for electroporation and transient expression of foreign genes in sugar-cane protoplasts. Plant Sci 81: 65–74 (1992).

    Google Scholar 

  31. Rathus C, Birch RG: Stable transformation of callus from electroporated sugar-cane protoplasts. Plant Sci 82: 81–89 (1992).

    Google Scholar 

  32. Vasil V, Castillo AM, Fromm ME, Vasil IK: Herbicide resistant fertile transgenic wheat plants obtained by microprojectile bombardment of regenerable embryogenic callus. Bio/technology 10: 667–674 (1992).

    Google Scholar 

  33. Walker JC, Howard EA, Dennis ES, Peacock WJ: DNA sequences required for anaerobic expression of the maize alcohol dehydrogenase 1 gene. Proc Natl Acad Sci USA 84: 6624–6628 (1987).

    Google Scholar 

  34. Walters DA, Vetsch CS, Potts DE, Lundquist RC: Transformation and inheritance of a hygromycin phosphotransferase gene in maize plants. Plant Mol Biol 18: 189–200 (1992).

    Google Scholar 

Download references

Author information

Author notes

  1. Carl Rathus

    Present address: CSIRO Division of Tropical Crops and Pastures, Brisbane, Australia

Authors and Affiliations

  1. Bureau of Sugar Experiment Stations, Brisbane, Australia

    Carl Rathus

  2. Department of Botany, University of Queensland, 4072, Australia

    Robert Bower & Robert G. Birch

Authors
  1. Carl Rathus
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Robert Bower
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Robert G. Birch
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rathus, C., Bower, R. & Birch, R.G. Effects of promoter, intron and enhancer elements on transient gene expression in sugar-cane and carrot protoplasts. Plant Mol Biol 23, 613–618 (1993). https://doi.org/10.1007/BF00019308

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00019308

Key words

Search

Navigation