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Production of spider silk proteins in tobacco and potato

Nature Biotechnology volume 19, pages 573–577 (2001)Cite this article

Abstract

Spider dragline silk is a proteinaceous fiber with remarkable mechanical properties that make it attractive for technical applications. Unfortunately, the material cannot be obtained in large quantities from spiders. We have therefore generated transgenic tobacco and potato plants that express remarkable amounts of recombinant Nephila clavipes dragline proteins. Using a gene synthesis approach, the recombinant proteins exhibit homologies of >90% compared to their native models. Here, we demonstrate the accumulation of recombinant silk proteins, which are encoded by synthetic genes of 420–3,600 base pairs, up to a level of at least 2% of total soluble protein in the endoplasmic reticulum (ER) of tobacco and potato leaves and potato tubers, respectively. Using the present expression system, spider silk proteins up to 100 kDa could be detected in plant tissues. When produced in plants, the recombinant spidroins exhibit extreme heat stability—a property that is used to purify the spidroins by a simple and efficient procedure.

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Figure 1: Basic elements for synthetic spider silk genes.
Figure 2: Synthetic spidroins and plant expression cassettes.
Figure 3: Detection of synthetic spider silk proteins in transgenic tobacco and potato plant organs.
Figure 4: Solubility, heat stability, and acid solubility of plant-produced synthetic silk proteins from tobacco leaves.

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References

  1. Tirrell, D.A. Putting a new spin on spider silk. Science 271, 39–40 (1996).

    Article  CAS  Google Scholar 

  2. Hinman, M.B., Jones, J.A. & Lewis R.V. Synthetic spider silk: a modular fibre. TIBTECH 18, 374–379 (2000).

    Article  CAS  Google Scholar 

  3. Gosline, J.M., Guerette, P.A., Ortlepp, C.S. & Savage, K.N. The mechanical design of spider silks: from fibroin sequence to mechanical function. J. Exp. Biol. 23, 3295–3303 (1999).

    Google Scholar 

  4. Hood, E.E. & Jilka, J.M. Plant-based production of xenogenic proteins. Curr. Opin. Biotechnol. 10, 382–386 (2000).

    Article  Google Scholar 

  5. Fiedler, U., Phillips, J., Artsaenko, O. & Conrad, U. Optimization of scFv production in transgenic plants. Immunotechnology 3, 205–216 (1997).

    Article  CAS  Google Scholar 

  6. Munro, S. & Pelham, H.R. A C-terminal signal prevents secretion of luminal ER proteins. Cell 48, 899–907 (1987).

    Article  CAS  Google Scholar 

  7. Saito, K., Keneko, H., Yyamazaki, M., Yoshida, M. & Murakoshi, I. Stable transfer and expression of chimeric genes in licorice (Glycyrrhiza uralensis) using an Ri plasmid binary vector. Plant Cell Rep. 6, 718–721 (1990).

    Article  Google Scholar 

  8. Bevan, M. Binary Agrobacterium vectors for plant transformation. Nucleic Acid Res. 12, 8711–8721 (1984).

    Article  CAS  Google Scholar 

  9. Fahnestock, S.R. & Irwin, S.L. Synthetic spider dragline silk proteins and their production in Escherichia coli. Appl. Microbiol. Biotechnol. 47, 23–32 (1997).

    Article  CAS  Google Scholar 

  10. Wandelt, C.I. et al. Vicilin with carboxy-terminal KDEL is retained in the endoplasmic reticulum and accumulates to high levels in the leaves of transgenic plants. Plant J. 2, 181–192 (1992).

    CAS  PubMed  Google Scholar 

  11. Prince, J.T., McGrath, K.P., DiGirolamo, C.M. & Kaplan, D.L. Construction, cloning, and expression of synthetic genes encoding spider dragline silk. Biochemistry 34, 10879–10885 (1995).

    Article  CAS  Google Scholar 

  12. Artsaenko, O. et al. Expression of a single-chain Fv antibody against abscisic acid creates a wilty phenotype in transgenic tobacco. Plant J. 8, 745–750 (1995).

    Article  CAS  Google Scholar 

  13. Zambriski, P. et al. Ti-plasmid vector for introduction of DNA into plant cells without altering their normal regeneration capacity. EMBO J. 2, 2143–2150 (1983).

    Article  Google Scholar 

  14. Conrad, U., Fiedler, U., Artsaenko, O. & Phillips, J. Single-chain Fv antibodies expressed in plants. In Methods in biotechnology—recombinant proteins from plants: production and isolation of clinically useful compounds. (eds Cunningham, C. & Porter, A.) 103–127 (Humana Press, Totowa, NJ; 1997).

    Google Scholar 

Download references

Acknowledgements

The authors thank K. Heinemann (Rudolstadt) for initial ideas and discussions, and I. Tiroke, I. Pfort, and C. Helmold for excellent technical assistance. This study was supported by the Bundesministerium für Landwirtschaft und Forsten represented by Fachagentur für Nachwachsende Rohstoffe (98NR050).

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Authors and Affiliations

  1. Institut für Pflanzengenetik und Kulturpflanzenforschung Gatersleben, Corrensstraße 3, Gatersleben, D-06466, Germany

    Jürgen Scheller & Udo Conrad

  2. Institut für Molekulare Biotechnologie e. V., PF 100 813, Jena, D-07708, Germany

    Karl-Heinz Gührs & Frank Grosse

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  1. Jürgen Scheller
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  2. Karl-Heinz Gührs
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  3. Frank Grosse
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  4. Udo Conrad
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Correspondence to Udo Conrad.

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Scheller, J., Gührs, KH., Grosse, F. et al. Production of spider silk proteins in tobacco and potato. Nat Biotechnol 19, 573–577 (2001). https://doi.org/10.1038/89335

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