Skip to main content
Log in

Promoter/leader deletion analysis and plant expression vectors with the figwort mosaic virus (FMV) full length transcript (FLt) promoter containing single or double enhancer domains

  • Published:
Transgenic Research Aims and scope Submit manuscript

Abstract

The boundaries required for maximal expression from the promoter/leader region of the full length transcript of figwort mosaic virus (FLt promoter) coupled to reporter genes were defined by 5′ and 3′ deletion analyses. In transient expression assays using protoplasts of Nicotiana edwardsonii, a 314 bp FLt promoter fragment sequence (−249 to +65 from the transcription start site) was sufficient for strong expression activity. Plant expression vectors developed with modified FLt promoters were tested with GUS or CAT as reporter genes in transgenic plants. The FLt promoter is a strong constitutive promoter, with strength comparable to or greater than that of the CaMV 35S promoter. The FLt promoter with its double enhancer domain linked to GUS or CAT reporter genes provides an average 4- fold greater activity than the FLt promoter with a single enhancer domain (−55 to −249 bp upstream fragment) in tests with transgenic plants and in protoplast transient expression assays

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  • An, G., Ebert, P.R., Yi, B.Y. and Choi, C-H. (1986) Both TATA box and upstream regions are required for nopaline synthase promoter activity in transformed tobacco cells. Mol. Gen. Genet. 203, 245-50.

    Article  Google Scholar 

  • An, G., Costa, M.A., Mitra, A., Ha, S.B. and Marton, L. (1988) Organ-specific and developmental regulation of the nopaline synthase promoter in transgenic tobacco plants. Plant Physiol. 88, 547-52.

    Google Scholar 

  • Assaad, F.F. and Signer, E.R. (1990) Cauliflower mosaic virus P35S promoter activity in Escherichia coli. Mol. Gen. Genet. 223, 517-520.

    Google Scholar 

  • Baughman, G. and Howell, S.H. (1988) Cauliflower mosaic virus 35S RNA leader region inhibits translation of downstream genes. Virology 167, 125-35.

    Article  PubMed  Google Scholar 

  • Benfey, P.N. and Chua, N.H. (1990) The cauliflower mosaic virus 35S promoter: Combinatorial regulation of transcription in plants. Science 250, 959-66.

    Google Scholar 

  • Benfey, P.N., Ren, L. and Chua, N.H. (1989) The CaMV 35S enhancer contain at least two domains which can confer different developmental and tissue-specific expression patterns. EMBO J. 8, 2195-202.

    Google Scholar 

  • Benfey, P.N., Ren, L. and Chua, N.H. (1990a) Combinatorial and synergistic properties of CaMV 35S enhancer subdomains. EMBO J. 9, 1685-96.

    PubMed  Google Scholar 

  • Benfey, P.N., Ren, L. and Chua, N.H. (1990b) Tissue-specific expression from CaMV 35S enhancer subdomains in early stages of plant development. EMBO J. 9, 1677-84.

    PubMed  Google Scholar 

  • Bienz, M. and Pelham, H.R.B. (1986) Heat shock regulatory elements function as an inducible enhancer in the Xenopus hsp 70gene and when linked to a heterologous promoter. Cell 45, 753-60.

    Article  PubMed  Google Scholar 

  • Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248-54.

    Article  PubMed  Google Scholar 

  • Day Dowson, M.J., Ashurst, J.L., Mathias, S.F., Watts, J.W., Wilson, T.M.A. and Dixon, R.A. (1993) Plant viral leaders influence expression of a reporter gene in tobacco. Plant Mol. Biol. 23, 97-109.

    PubMed  Google Scholar 

  • Driesen, M., Benito-Moreno, R.M., Hohn, T. and Futterer, J. (1993) Transcription from the CaMV 19S promoter and autocatalysis of translation from CaMV RNA. Virology 195, 203-10.

    Article  PubMed  Google Scholar 

  • Ellis, J.G., Llewellyn, D.J., Walker, J.C., Dennis, E.S. and Peacock, W.J. (1987) The OCS-elements: a 16 base pair palindrome essential for activity of the octopine synthase enhancer. EMBO J. 6, 3203-8.

    Google Scholar 

  • Fang, R-X., Nagy, F., Sivasubramanian, S. and Chua, N.H. (1989) Multiple cisregulatory elements for maximal expression of the cauliflower mosaic virus 35S promoter in transgenic plants. Plant Cell 1, 141-50.

    Article  PubMed  Google Scholar 

  • Fromental, C., Kanno, M., Nomiyama, H. and Chambon, P. (1988) Cooperativity and hierarchical levels of functional organization in the SV 40 enhancer. Cell 54, 943-53.

    Article  PubMed  Google Scholar 

  • Gardner, R.C., Howarth, A.J., Hahn, P., Brown-Leudi, M. and Shepherd, R.J. (1981) The complete nucleotide sequence of an infectious clone of cauliflower mosaic virus by M13mp7 shortgun sequencing. Nucl. Acids Res. 9, 2871-88.

    PubMed  Google Scholar 

  • Gorman, C.M., Moffat, L.F. and Howard, B.H. (1982) Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol. Cell Biol. 2, 1044-51.

    PubMed  Google Scholar 

  • Gowda, S., Wu, F.C., Herman, H.B. and Shepherd, R.J. (1989) Gene VI of figwort mosaic virus (caulimovirus group) functions in posttranscriptional expression of genes on the full-length RNA transcript. Proc. Natl Acad. Sci. USA 86, 9203-7.

    PubMed  Google Scholar 

  • Guilley, H., Dudley, R.K., Jonard, G., Balazs, E. and Richards, K.E. (1982) Transcription of cauliflower mosaic virus DNA: detection of promoter sequences, and characterization of transcripts. Cell 30, 763-73.

    Article  PubMed  Google Scholar 

  • Hasegawa, A., Verver, J., Shimada, A., Saito, M., Goldbach, R., van Kammen, A., Miki, K., Kameya-Iwaki, M. and Hibi, T. (1989) The complete sequence of soybean chlorotic mottle virus DNA and the identification of a novel promoter. Nucl. Acids Res. 17, 9993-10013.

    PubMed  Google Scholar 

  • Howell, S.H. and Hull, R. (1978) Replication of cauliflower mosaic virus and transcription of its genome in turnip leaf protoplasts. Virology 86, 468-81.

    Article  PubMed  Google Scholar 

  • Hull, R., Sadler, J. and Longstaff, M. (1986) The sequence of carnation etched ring viral DNA: comparison with cauliflower mosaic virus and retroviruses. EMBO J. 5, 3083-90.

    Google Scholar 

  • Jefferson, R.A., Kananagh, T.A. and Bevan, M.W. (1987) GUS fusion: ·-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6, 3901-7.

    PubMed  Google Scholar 

  • Kay, R., Chan, R., Daly, M. and McPherson, J. (1987) Duplication of CaMV 35S promoter sequence creats a strong enhancer for plant genes. Science 236, 1299-1302.

    Google Scholar 

  • Khoury, G. and Gruss, P. (1983) Enhancer elements. Cell 33, 313- 4.

    Article  PubMed  Google Scholar 

  • Kiernan, J.M., Wu, F.C., Goldberg, K.B., Gowda, S. and Shepherd, R.J. (1993) Transformation in Nicotiana edwardsonii, In Bajaj, Y.P.S. ed., Biotechnology in Agriculture and Forestry, Vol 22, Plant Protoplasts and Genetic Engineering III, pp. 294-307, Springer-Verlag Berlin Heidelberg.

    Google Scholar 

  • Lam, E. (1994) Analysis of tissue-specific elements in the CaMV 35S promoter. In Nover, L. ed, Results and Problems in Cell Differentiation, Plant Promoters and transcription factors, Vol 20, pp. 181-96, Springer-Verlag Berlin Heidelberg.

    Google Scholar 

  • Lam, E. and Chua, N.H. (1989) ASF-2: A factor that binds to the cauliflower mosaic virus 35S promoter and a conserved GATA motif in Cab promoters. Plant Cell 1, 1147-56.

    Article  PubMed  Google Scholar 

  • Lam, E., Benfey, P.N., Gilmartin, P.M., Fang, R.X. and Chua, N.H. (1989) Site-specific mutations alter in vitrofactor binding and change promoter expression pattern in transgenic plants. Proc. Natl Acad. Sci. USA 86, 7890-4.

    PubMed  Google Scholar 

  • Lawton, M.A., Tierney, M.A., Nakamura, I., Anderson, E., Komeda, Y., Dube, P., Hoffman, N., Fraley, R.T. and Beachy, R.N. (1987) Expression of a soybean 25CF-conglycinin gene under the control of the cauliflower mosaic virus 35S and 19S promoters in transformed petunia tissues. Plant Mol. Biol. 9, 315-24.

    Google Scholar 

  • Maiti, I.B., Murphy, J.F., Shaw, J.G. and Hunt, A.G. (1993) Plant that express a potyvirus proteinase genes are resistant to virus infection. Proc. Natl Acad. Sci. USA 90, 6110-4.

    PubMed  Google Scholar 

  • Medberry, S.L., Lockhart, B.E.L. and Olszewski, N.E. (1992) The commelina yellow mottle virus promoter is a strong promoter in vascular and reproductive tissue. Plant Cell 4: 185-92.

    Article  PubMed  Google Scholar 

  • Morelli, G., Nagy, F., Fraley, R.T., Rogers, S.G. and Chua, N.H. (1985) A short conserved sequence is involved in the light inducibility of a gene encoding ribulose-1, 5-bisphosphate carboxylase small subunit of pea. Nature 315, 200-4.

    Google Scholar 

  • Odell, J.T., Dudley, R.K. and Howell, S.H. (1981) Structure of the 19S RNA transcripts encoded by the cauliflower mosaic virus genome. Virology 111, 377-385.

    Article  Google Scholar 

  • Odell, J.T., Nagy, F. and Chua, N.H. (1985) Identification of DNA sequence required for activity of the cauliflower mosaic virus Figwort mosaic virus promoter analysis155 35S promoter. Nature 313, 810-2.

    PubMed  Google Scholar 

  • Odell, J.T., Knowlton, S., Lin, W. and Mauvais, C.J. (1988) Properties of an isolated transcription stimulating sequence derived from the cauliflower mosaic virus 35S promoter. Plant Mol. Biol. 10, 263-72.

    Google Scholar 

  • Omirulleh, S., Abraham, M., Golovkin, M., Stefanov, I., Karabaev, M.K., Mustardy, L., Morocz, S. and Dudits, D. (1993) Activity of a chimeric promoter with the double CaMV 35S enhancer elements in protoplast-derived cells and transgenic plants in maize. Plant Mol. Biol. 21, 415-28.

    PubMed  Google Scholar 

  • Ondek, B., Gloss, L. and Herr, W. (1988) The SV 40 enhancer contains two distinct levels of organization. Nature 333, 40-5.

    Article  PubMed  Google Scholar 

  • Ow, D.W., Jacobs, J.D. and Howell, S.H. (1987) Functional region of the cauliflower mosaic virus 35S RNA promoter determined by the use of the firefly luciferase gene as a reporter of promoter activity. Proc. Natl Acad. Sci. USA 84, 4870-4.

    Google Scholar 

  • Pobjecky, N., Rosenberg, G.H., Dinter-Gottlieb, G. and Kaufer, N.F. (1990) Expression of the 25CF-glucuronidase gene under the control of the CaMV 35S promoter in Schizosaccharomyces pombe. Mol. Gen. Genet. 220, 314-6.

    Article  PubMed  Google Scholar 

  • Richins, R.D. (1993) Organization and expression of the peanut chlorotic streak virus genome. Ph.D. Dissertation, at the University of Kentucky, Lexington, Kentucky, USA.

    Google Scholar 

  • Richins, R.D., Scholthof, H.B. and Shepherd, R.J. (1987) Sequence of figwort mosaic virus DNA (caulimovirus group). Nucl. Acids Res. 15, 8451-66.

    PubMed  Google Scholar 

  • Richins, R.D., Broos, T., Ducasse, D.A., Gowda, S., Mushegian, A.R., Reddy, D.V.R. and Shepherd, R.J. (1994) The complete nucleotide sequence of peanut chlorotic streak virus (PClSV) DNA. EMBL Data Library Gene Bank accession No. U13988.

  • Sanger, F., Nicklen, S. and Coulson, A.R. (1977) DNA sequencing with chain terminating inhibitors. Proc. Natl Acad. Sci. USA 74, 5463-7.

    PubMed  Google Scholar 

  • Sanger, M., Daubert, S. and Goodman, R.M. (1990) Characteristics of a strong promoter from figwort mosaic virus: comparison with the analogous 35S promoter from cauliflower mosaic virus and the regulated mannopine synthase promoter. Plant Mol. Biol. 14, 433-43.

    PubMed  Google Scholar 

  • Schardl, C.L., Byrd, A.D., Benzion, G., Altschuler, M.A., Hildebrand, D.F. and Hunt, A.G. (1987) Design and construction of a versatile system for the expression of foreign genes in plants. Gene 61, 1-11.

    PubMed  Google Scholar 

  • Schirm, S. and Jiricny, J. and Schaffner, W. (1987) The SV40 enhancer can be dissected into multiple segments, each with a different cell type specificity. Genes. Dev. 1, 65-74.

    PubMed  Google Scholar 

  • Scholthof, H.B., Gowda, S., Wu, F. and Shepherd, R.J. (1992) The full-length transcript of a caulimovirus is a polycistronic mRNA whose genes are transactivated by the product of gene VI. J. Virol. 66, 3131-9.

    PubMed  Google Scholar 

  • Shepherd, R.J., Richins, R.D., Duffus, J.E., and Handley, M.K. (1987) Figwort mosaic virus: properties of the virus and its adaptation to a new host. Phytopathology 77, 1668-73.

    Google Scholar 

  • Thomson, D., and Henry, R. (1993) Use of DNA from dry leaves for PCR and RAPD analysis. Plant Mol. Biol. Rep. 11, 202-6.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Maiti, I.B., Gowda, S., Kiernan, J. et al. Promoter/leader deletion analysis and plant expression vectors with the figwort mosaic virus (FMV) full length transcript (FLt) promoter containing single or double enhancer domains. Transgenic Res 6, 143–156 (1997). https://doi.org/10.1023/A:1018477705019

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1018477705019

Navigation