Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1987 Nov;6(11):3203–3208. doi: 10.1002/j.1460-2075.1987.tb02636.x

The ocs element: a 16 base pair palindrome essential for activity of the octopine synthase enhancer

J G Ellis 1, D J Llewellyn 1, J C Walker 1, E S Dennis 1, W J Peacock 1
PMCID: PMC553770  PMID: 16453801

Abstract

A 176 bp DNA sequence lying upstream of the octopine synthase (ocs) promoter, previously shown to have enhancer-like properties in transgenic tobacco [Ellis et al. (1987) EMBO J., 6, 11-16], functions as an enhancer in protoplasts of Zea mays (a monocot plant) and Nicotiana plumbaginifolia (a dicotplant). We have characterized this element by transient expression assays using a linked alcohol dehydrogenase (Adh1) promoter from Z. mays and the chloramphenicol acetyltransferase coding sequences. The ocs sequence functions in both orientations but its enhancing activity is dependent upon its distance from the Adh1 promoter. Transient expression assays using deletion mutants and synthetic oligonucleotides show that a 16 bp palindrome ACGTAAGCGCTTACGT, contained within the 176 bp fragment, is essential and sufficient for enhancing activity in transient expression assays.

Keywords: octopine synthase enhancer element, Adh1 promoter, transcription signals, plant protoplasts

Full text

PDF
3203

Images in this article

3204Fig. 2.
on p.3204
3205Fig. 4.
on p.3205
3206Fig. 5.
on p.3206

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Banerji J., Rusconi S., Schaffner W. Expression of a beta-globin gene is enhanced by remote SV40 DNA sequences. Cell. 1981 Dec;27(2 Pt 1):299–308. doi: 10.1016/0092-8674(81)90413-x. [DOI] [PubMed] [Google Scholar]
  2. Bevan M., Barnes W. M., Chilton M. D. Structure and transcription of the nopaline synthase gene region of T-DNA. Nucleic Acids Res. 1983 Jan 25;11(2):369–385. doi: 10.1093/nar/11.2.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bienz M., Pelham H. R. Heat shock regulatory elements function as an inducible enhancer in the Xenopus hsp70 gene and when linked to a heterologous promoter. Cell. 1986 Jun 6;45(5):753–760. doi: 10.1016/0092-8674(86)90789-0. [DOI] [PubMed] [Google Scholar]
  4. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  5. De Block M., Herrera-Estrella L., Van Montagu M., Schell J., Zambryski P. Expression of foreign genes in regenerated plants and in their progeny. EMBO J. 1984 Aug;3(8):1681–1689. doi: 10.1002/j.1460-2075.1984.tb02032.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. De Greve H., Dhaese P., Seurinck J., Lemmers M., Van Montagu M., Schell J. Nucleotide sequence and transcript map of the Agrobacterium tumefaciens Ti plasmid-encoded octopine synthase gene. J Mol Appl Genet. 1982;1(6):499–511. [PubMed] [Google Scholar]
  7. Dennis E. S., Gerlach W. L., Pryor A. J., Bennetzen J. L., Inglis A., Llewellyn D., Sachs M. M., Ferl R. J., Peacock W. J. Molecular analysis of the alcohol dehydrogenase (Adh1) gene of maize. Nucleic Acids Res. 1984 May 11;12(9):3983–4000. doi: 10.1093/nar/12.9.3983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Depicker A., Stachel S., Dhaese P., Zambryski P., Goodman H. M. Nopaline synthase: transcript mapping and DNA sequence. J Mol Appl Genet. 1982;1(6):561–573. [PubMed] [Google Scholar]
  9. Ellis J. G., Llewellyn D. J., Dennis E. S., Peacock W. J. Maize Adh-1 promoter sequences control anaerobic regulation: addition of upstream promoter elements from constitutive genes is necessary for expression in tobacco. EMBO J. 1987 Jan;6(1):11–16. doi: 10.1002/j.1460-2075.1987.tb04711.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Freeman G. J., Clayberger C., DeKruyff R., Rosenblum D. S., Cantor H. Sequential expression of new gene programs in inducer T-cell clones. Proc Natl Acad Sci U S A. 1983 Jul;80(13):4094–4098. doi: 10.1073/pnas.80.13.4094. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fromm M., Berg P. Simian virus 40 early- and late-region promoter functions are enhanced by the 72-base-pair repeat inserted at distant locations and inverted orientations. Mol Cell Biol. 1983 Jun;3(6):991–999. doi: 10.1128/mcb.3.6.991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fromm M., Taylor L. P., Walbot V. Expression of genes transferred into monocot and dicot plant cells by electroporation. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5824–5828. doi: 10.1073/pnas.82.17.5824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Grosveld G. C., de Boer E., Shewmaker C. K., Flavell R. A. DNA sequences necessary for transcription of the rabbit beta-globin gene in vivo. Nature. 1982 Jan 14;295(5845):120–126. doi: 10.1038/295120a0. [DOI] [PubMed] [Google Scholar]
  14. Hill D. E., Hope I. A., Macke J. P., Struhl K. Saturation mutagenesis of the yeast his3 regulatory site: requirements for transcriptional induction and for binding by GCN4 activator protein. Science. 1986 Oct 24;234(4775):451–457. doi: 10.1126/science.3532321. [DOI] [PubMed] [Google Scholar]
  15. Koncz C., De Greve H., André D., Deboeck F., Van Montagu M., Schell J. The opine synthase genes carried by Ti plasmids contain all signals necessary for expression in plants. EMBO J. 1983;2(9):1597–1603. doi: 10.1002/j.1460-2075.1983.tb01630.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Llewellyn D. J., Finnegan E. J., Ellis J. G., Dennis E. S., Peacock W. J. Structure and expression of an alcohol dehydrogenase 1 gene from Pisum sativum (cv. "Greenfeast"). J Mol Biol. 1987 May 5;195(1):115–123. doi: 10.1016/0022-2836(87)90331-7. [DOI] [PubMed] [Google Scholar]
  17. Otten L., De Greve H., Hernalsteens J. P., Van Montagu M., Schieder O., Straub J., Schell J. Mendelian transmission of genes introduced into plants by the Ti plasmids of Agrobacterium tumefaciens. Mol Gen Genet. 1981;183(2):209–213. doi: 10.1007/BF00270619. [DOI] [PubMed] [Google Scholar]
  18. Parker C. S., Topol J. A Drosophila RNA polymerase II transcription factor binds to the regulatory site of an hsp 70 gene. Cell. 1984 May;37(1):273–283. doi: 10.1016/0092-8674(84)90323-4. [DOI] [PubMed] [Google Scholar]
  19. Timko M. P., Kausch A. P., Castresana C., Fassler J., Herrera-Estrella L., Van den Broeck G., Van Montagu M., Schell J., Cashmore A. R. Light regulation of plant gene expression by an upstream enhancer-like element. Nature. 1985 Dec 12;318(6046):579–582. doi: 10.1038/318579a0. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES