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Novel and useful properties of a chimeric plant promoter combining CaMV 35S and MAS elements

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Abstract

The CaMV 35S and Ti plasmid mannopine synthetase (mas) promoters are commonly used by plant genetic engineers. To combine their useful properties, we constructed hybrid promoters incorporating elements from both. These promoters were spliced to the beta-glucuronidase reporter gene and introduced into tobacco and tomato plants by Agrobacterium cocultivation. T1 and T2 transgenic plant populations transformed with different constructs were assayed for the marker enzyme. Comparisons were made based on the range of expression levels found for each promoter construct. We found that a hybrid promoter incorporating the mas region from +65 to -301 and the 35S enhancer region from -90 to -941 had new and interesting properties. This promoter, called Mac, expressed gus at a level three to five times that expressed by a double 35S promoter in the leaves, and 10 to 15 times in hypocotyls and roots. The Mac promoter, however, showed only marginal wound inducibility. Five- to seven-fold wound induction required the presence of the region from -301 to -613 of mas. Reiteration of the 35S enhancer region, from -90 to -430, behind the 35S TATA box region or the mas +65 to -301 region had a smaller effect on expression, ranging from equal to twice the level of the single enhancer control.

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References

  1. Atchison M: Enhancers: mechanism of action and cell specificity. Ann Rev Cell Biol 4: 127–153 (1988).

    PubMed  Google Scholar 

  2. Barker RF, Idler KB, Thompson DV, Kemp JD: Nucleotide sequence of the T-DNA region from the Agrobacterium tumefaciens octopine Ti plasmid pTi15955. Plant Mol Biol 2: 335–350 (1983).

    Google Scholar 

  3. Bienz M, Pelham HRB: Heat shock regulatory element function as an independent enhancer in the Xenopus HSP70 gene and when linked to a heterologous promoter. Cell 45: 753–760 (1986).

    Article  PubMed  Google Scholar 

  4. Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72: 248–254 (1976).

    Article  PubMed  Google Scholar 

  5. Comai L, Facciotti D, Hiatt WR, Thompson G, Rose RE, Stalker DM: Expression in plants of a mutant aroA gene from Salmonella typhymurium confers tolerance to glyphosate. Nature 317: 741–744 (1985).

    Google Scholar 

  6. DiRita VJ, Gelvin SB: Deletion analysis of the mannopine synthase gene promoter in sunflower crown gall tumors and Agrobacterium tumefaciens. Mol Gen Genet 207: 233–241 (1987).

    Google Scholar 

  7. Ellis JG, Llewellyn DJ, Dennis ES, Peacock WJ: Maize ADH-1 promoter sequences control anaerobic regulation: addition of upstream promoter elements from constitutive genes is necessary for expression in tobacco. EMBO J 6: 11–16 (1987).

    Google Scholar 

  8. Fang R-X, Nagy F, Jivasubramanian AM, 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).

    Article  PubMed  Google Scholar 

  9. Fillatti JJ, Kiser J, Rose R, Comai L: Efficient transfer of a glyphosate tolerance gene into tomato using a binary Agrobacterium tumefaciens vector. Bio/Technology 5: 725–730 (1987).

    Article  Google Scholar 

  10. Franck A, Guilley H, Jonard G, Richards K, Hirth: Nucleotide sequence of Cauliflower Mosaic Virus DNA. Cell 21: 285–294 (1980).

    Article  PubMed  Google Scholar 

  11. Gallie DR, Sleat DE, Watts JW, Turner PC, Wilson TMA: A comparison of eukaryotic viral 5′ leader sequences as enhancers of mRNA expression in vivo. Nucleic Acid Res 15: 8693–8711 (1987).

    PubMed  Google Scholar 

  12. Gardner RC, Howarth AJ, Hahn P, Brown-Luedi M, Shepherd RJ, Messing J: The complete nucleotide sequence of an infectious clone of Cauliflower Mosaic Virus by M13mp7 shotgun cloning. Nucl Acids Res 9: 2871–2889 (1981).

    PubMed  Google Scholar 

  13. Harpster MH, Townsend JA, Jones JDG, Bedbrook J, Dunsmuir P: Relative strengths of the 35S cauliflower mosaic virus, 1′, 2′, and nopaline synthase promoters in transformed tobacco, sugarbeet, and oilseed rape callus tissue. Mol Gen Genet 212: 182–190 (1988).

    PubMed  Google Scholar 

  14. Hoekema A, Hirsch PR, Hooykaas PJJ, Schilperoort RA: A binary vector based on separation of vir and T-region of the Agrobacterium tumefaciens Ti plasmids. Nature 303: 179–181 (1983).

    Google Scholar 

  15. Horsch RB, Fry JE, Hoffman NL, Eichholtz D, Rogers SG, Fraley RT: A simple and general method for transferring genes into plants. Science 227: 1229–1231 (1985).

    Google Scholar 

  16. Jefferson RA, Kavanagh TA, Bevan MW: Gus fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6: 3901–3907 (1987).

    PubMed  Google Scholar 

  17. Karcher SJ, DiRita VJ, Gelvin SB: Transcript analysis of Tr-DNA in octopine-type crown gall tumors. Mol Gen Genet 194: 159–165 (1984).

    Google Scholar 

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

    Google Scholar 

  19. Langridge WHR, Fitzgerald KJ, Koncz C, Schell J, Szalay AA: Dual promoter of Agrobacterium tumefaciens mannopine synthase genes is regulated by plant growth hormones. Proc Natl Acad Sci USA 86: 3219–3223 (1989).

    Google Scholar 

  20. Leisner SM, Gelvin SB: Multiple domains exist within the upstream activator sequence of the octopine synthase gene. Plant Cell (1989).

  21. Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: a Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y. (1982).

    Google Scholar 

  22. McBride KE, Summerfelt KR: Improved binary vectors for Agrobacterium mediated plant transformation. Plant Mol Biol 14: 269–276 (1990).

    PubMed  Google Scholar 

  23. Odell JT, Knowlton S, Lin W, Mauvais CJ: Properties of an isolated transcription stimulating sequence derived from the cauliflower mosaic virus 35S promoter. Plant Mol Biol 10: 263–272 (1988).

    Google Scholar 

  24. Sanger M, Daubert S, Goodman RM: Characterization of a strong promoter from Figwart Mosaic Virus, comparison with the analogous 35S promoter from Cauliflower Mosaic Virus and the regulated mannopine synthase promoter. Plant Mol Biol 14: 433–443 (1990).

    PubMed  Google Scholar 

  25. Strittmatter G, Chua N-H: Artificial combination of two cis-regulatory elements generates a unique pattern of expression in transgenic plants. Proc Natl Acad Sci USA 84: 8986–8990 (1987). lac

    PubMed  Google Scholar 

  26. Teeri TH, Lehvaslaiho H, Frank M, Uotila J, Heino P, Palva ET, VanMontagu M, Herrera-Estrella L: Gene fusions to lacZ reveal new expression patterns of chimeric genes in transgenic plants. EMBO J 8: 343–350 (1989).

    PubMed  Google Scholar 

  27. Velten J, Velten J, Hain R, Schell J: Isolation of a dual plant promoter fragment from the Ti plasmid of Agrobacterium tumefaciens. EMBO J 3: 2723–2730 (1984).

    Google Scholar 

  28. Winter JA, Wright RL, Gurley WB: Map locations of five transcripts homologous to Tr-DNA in tobacco and sunflower crown gall tumors. Nucl Acids Res 12: 2391–2406 (1984).

    PubMed  Google Scholar 

  29. Yanish-Perron C, Vieira J, Messing J: Improved M13 cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33: 103–119 (1985).

    Article  PubMed  Google Scholar 

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Comai, L., Moran, P. & Maslyar, D. Novel and useful properties of a chimeric plant promoter combining CaMV 35S and MAS elements. Plant Mol Biol 15, 373–381 (1990). https://doi.org/10.1007/BF00019155

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  • DOI: https://doi.org/10.1007/BF00019155

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