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. 1987 Dec 1;6(12):3559–3564. doi: 10.1002/j.1460-2075.1987.tb02685.x

Localization of sequences in wheat endosperm protein genes which confer tissue-specific expression in tobacco

V Colot 1, L S Robert 1,1, T A Kavanagh 1,2, M W Bevan 1, R D Thompson 1,3
PMCID: PMC553821  PMID: 15467781

Abstract

The developing cereal grain accumulates large quantities of proteins which are unique to the endosperm tissue. The DNA sequences which determine their endosperm-specific expression have not yet been identified. In the absence of a suitable transformation-regeneration system for cereals, we have investigated whether chimaeric genes consisting of low mol. wt (LMW) and high mol. wt (HMW) glutenin gene upstream sequences coupled to the coding region of the bacterial chloramphenicol acetyl transferase (CAT) gene could be specifically expressed in transgenic tobacco. The fusions, made in a Ti-derived binary vector, were introduced into tobacco via Agrobacterium tumefaciens-mediated transformation and their activity assayed. Both the LMW and HMW glutenin chimaeric genes exhibited endosperm-specific CAT activity in the transformed plants. In addition, a deletion series of the LMW glutenin sequence indicated that sequences present between 326 bp and 160 bp upstream of the transcription start point are necessary to confer endosperm-specific CAT activity.

Keywords: endosperm, glutenins, tissue-specific expression, transgenic tobacco, wheat promoter

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Selected References

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

  1. A simple and general method for transferring genes into plants. Science. 1985 Mar 8;227(4691):1229–1231. doi: 10.1126/science.227.4691.1229. [DOI] [PubMed] [Google Scholar]
  2. Anderson O. D., Litts J. C., Gautier M. F., Greene F. C. Nucleic acid sequence and chromosome assignment of a wheat storage protein gene. Nucleic Acids Res. 1984 Nov 12;12(21):8129–8144. doi: 10.1093/nar/12.21.8129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baker B., Schell J., Lörz H., Fedoroff N. Transposition of the maize controlling element "Activator" in tobacco. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4844–4848. doi: 10.1073/pnas.83.13.4844. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bevan M. W., Mason S. E., Goelet P. Expression of tobacco mosaic virus coat protein by a cauliflower mosaic virus promoter in plants transformed by Agrobacterium. EMBO J. 1985 Aug;4(8):1921–1926. doi: 10.1002/j.1460-2075.1985.tb03871.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bevan M. Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res. 1984 Nov 26;12(22):8711–8721. doi: 10.1093/nar/12.22.8711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  7. Czernilofsky A. P., Hain R., Herrera-Estrella L., Lörz H., Goyvaerts E., Baker B. J., Schell J. Fate of selectable marker DNA integrated into the genome of Nicotiana tabacum. DNA. 1986 Apr;5(2):101–113. doi: 10.1089/dna.1986.5.101. [DOI] [PubMed] [Google Scholar]
  8. Dynan W. S., Tjian R. Control of eukaryotic messenger RNA synthesis by sequence-specific DNA-binding proteins. 1985 Aug 29-Sep 4Nature. 316(6031):774–778. doi: 10.1038/316774a0. [DOI] [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. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  11. Fluhr R., Kuhlemeier C., Nagy F., Chua N. H. Organ-specific and light-induced expression of plant genes. Science. 1986 May 30;232(4754):1106–1112. doi: 10.1126/science.232.4754.1106. [DOI] [PubMed] [Google Scholar]
  12. Forde B. G., Kreis M., Williamson M. S., Fry R. P., Pywell J., Shewry P. R., Bunce N., Miflin B. J. Short tandem repeats shared by B- and C-hordein cDNAs suggest a common evolutionary origin for two groups of cereal storage protein genes. EMBO J. 1985 Jan;4(1):9–15. doi: 10.1002/j.1460-2075.1985.tb02310.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gillies S. D., Morrison S. L., Oi V. T., Tonegawa S. A tissue-specific transcription enhancer element is located in the major intron of a rearranged immunoglobulin heavy chain gene. Cell. 1983 Jul;33(3):717–728. doi: 10.1016/0092-8674(83)90014-4. [DOI] [PubMed] [Google Scholar]
  14. Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. doi: 10.1016/0378-1119(84)90153-7. [DOI] [PubMed] [Google Scholar]
  15. Herr W., Clarke J. The SV40 enhancer is composed of multiple functional elements that can compensate for one another. Cell. 1986 May 9;45(3):461–470. doi: 10.1016/0092-8674(86)90332-6. [DOI] [PubMed] [Google Scholar]
  16. Hu N. T., Peifer M. A., Heidecker G., Messing J., Rubenstein I. Primary structure of a genomic zein sequence of maize. EMBO J. 1982;1(11):1337–1342. doi: 10.1002/j.1460-2075.1982.tb01319.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kuhlemeier C., Fluhr R., Green P. J., Chua N. H. Sequences in the pea rbcS-3A gene have homology to constitutive mammalian enhancers but function as negative regulatory elements. Genes Dev. 1987 May;1(3):247–255. doi: 10.1101/gad.1.3.247. [DOI] [PubMed] [Google Scholar]
  18. Lamppa G., Nagy F., Chua N. H. Light-regulated and organ-specific expression of a wheat Cab gene in transgenic tobacco. Nature. 1985 Aug 22;316(6030):750–752. doi: 10.1038/316750a0. [DOI] [PubMed] [Google Scholar]
  19. Maier U. G., Brown J. W., Tologcyzki C., Feix G. Binding of a nuclear factor to a consensus sequence in the 5' flanking region of zein genes from maize. EMBO J. 1987 Jan;6(1):17–22. doi: 10.1002/j.1460-2075.1987.tb04712.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Maniatis T., Goodbourn S., Fischer J. A. Regulation of inducible and tissue-specific gene expression. Science. 1987 Jun 5;236(4806):1237–1245. doi: 10.1126/science.3296191. [DOI] [PubMed] [Google Scholar]
  21. Moreau P., Hen R., Wasylyk B., Everett R., Gaub M. P., Chambon P. The SV40 72 base repair repeat has a striking effect on gene expression both in SV40 and other chimeric recombinants. Nucleic Acids Res. 1981 Nov 25;9(22):6047–6068. doi: 10.1093/nar/9.22.6047. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Odell J. T., Nagy F., Chua N. H. Identification of DNA sequences required for activity of the cauliflower mosaic virus 35S promoter. 1985 Feb 28-Mar 6Nature. 313(6005):810–812. doi: 10.1038/313810a0. [DOI] [PubMed] [Google Scholar]
  23. Pedersen K., Devereux J., Wilson D. R., Sheldon E., Larkins B. A. Cloning and sequence analysis reveal structural variation among related zein genes in maize. Cell. 1982 Jul;29(3):1015–1026. doi: 10.1016/0092-8674(82)90465-2. [DOI] [PubMed] [Google Scholar]
  24. Rafalski J. A., Scheets K., Metzler M., Peterson D. M., Hedgcoth C., Söll D. G. Developmentally regulated plant genes: the nucleotide sequence of a wheat gliadin genomic clone. EMBO J. 1984 Jun;3(6):1409–1415. doi: 10.1002/j.1460-2075.1984.tb01985.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rahman S., Kreis M., Forde B. G., Shewry P. R., Miflin B. J. Hordein-gene expression during development of the barley (Hordeum vulgare) endosperm. Biochem J. 1984 Oct 15;223(2):315–322. doi: 10.1042/bj2230315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schirm S., Jiricny J., Schaffner W. The SV40 enhancer can be dissected into multiple segments, each with a different cell type specificity. Genes Dev. 1987 Mar;1(1):65–74. doi: 10.1101/gad.1.1.65. [DOI] [PubMed] [Google Scholar]
  27. Sengupta-Gopalan C., Reichert N. A., Barker R. F., Hall T. C., Kemp J. D. Developmentally regulated expression of the bean beta-phaseolin gene in tobacco seed. Proc Natl Acad Sci U S A. 1985 May;82(10):3320–3324. doi: 10.1073/pnas.82.10.3320. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  29. Thompson R. D., Bartels D., Harberd N. P. Nucleotide sequence of a gene from chromosome 1D of wheat encoding a HMW-glutenin subunit. Nucleic Acids Res. 1985 Oct 11;13(19):6833–6846. doi: 10.1093/nar/13.19.6833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. West R. W., Jr, Rodriguez R. L. Construction and characterization of E. coli promoter-probe plasmid vectors. III. pBR322 derivatives with deletions in the tetracycline resistance promoter region. Gene. 1982 Dec;20(2):291–304. doi: 10.1016/0378-1119(82)90047-6. [DOI] [PubMed] [Google Scholar]
  32. Zinn K., DiMaio D., Maniatis T. Identification of two distinct regulatory regions adjacent to the human beta-interferon gene. Cell. 1983 Oct;34(3):865–879. doi: 10.1016/0092-8674(83)90544-5. [DOI] [PubMed] [Google Scholar]

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