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US5279953: In vivo production of taxanes

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Filing Information

Inventor(s) Roy W. Stahlhut ·
Assignee(s) ESCA Genetics Corporation ·
Attorney/Agent(s) Jacques M. Dulin ·
Primary Examiner David T. Fox ·
Assistant Examiner Charles Rories ·
Application Number US7904371
Filing date 06/24/1992
Issue date 01/18/1994
Predicted expiration date 06/24/2012
U.S. Classifications 435/172.3  · 435/123  ·
International Classifications C12N 1500  ·
Kind CodeA
International Classifications 435156;170;123;41;240.4;240.48 ·
12 Claims, No Drawings


Abstract

Method for improved in vivo production of taxanes by inoculation of tissue of the genus Taxus with virulent or avirulent strains of Agrobacterium. In the preferred embodiment, virulent Agrobacterium is the inoculant which induces gall formation at all stages of the plant, from saplings to mature trees. Incomplete harvesting of galls permits regeneration on the trees, resulting in a renewable resource. Unexpectedly, the inoculant tissue (both virulent and avirulent) produces approximately twice the concentration of taxanes per unit tissue weight as compared to normal tissue. In addition, the virulent strain induced gall biomass is 2-3 times greater than normal tissue per unit growth time, while avirulent inoculants show approximately the same growth as normal. The net yield increase is 2-6 times normal, (4-6 for galls), is renewable, does not require special growth hormones or media, does not result in sacrificing the tree, and can be started at the sapling stage. The post-growth phase production is uncoupled by the inoculation and concentration yield is doubled.

Independent Claims | See all claims (12)

  1. 1. A method of in vivo production of taxanes comprising the steps of:a) inoculating a plant of the genus Taxus with Agrobacterium tumefaciens virulent strain A281 in at least one site on said plant;b) sustaining said plant for a time sufficient to induce said plant to respond with in vivo growth of gall tissue exhibiting production of secondary metabolites at the site of said inoculation;c) harvesting at least some of said gall tissue from said site; andd) extracting a taxane from said harvested gall tissue.
  2. 7. A method of in vivo production of taxanes comprising the steps of:a) inoculating a plant of the genus Taxus with Agrobacterium tumefaciens avirulent strain LBA4011 in at least one site on said plant;b) sustaining said plant for a time sufficient to induce said plant respond with growth of wound response tissue at the site of said inoculation, said wound response tissue exhibiting production of at least one taxane compound in excess of the production of surrounding normal, healthy plant tissue;c) harvesting at least some of said wound response tissue from said site; andd) extracting a taxane from said harvested wound response tissue.

References Cited

U.S. Patent Documents

Document NumberAssigneesInventorsIssue/Pub Date
US5019504 The United States of America as represented by the Secretary of Agriculture Christen et al. May 1991

Foreign Patent Documents

Document NumberAssigneesInventorsIssue/Pub Date
EP0224287Rijksuniversiteit LeidenMar 1987

Other Publications

E. Weiler et al. Trends in Biochem. Sci., vol. 12 (1987) pp. 271-275.
Luckner et al, Planta Medica 56:498-499 (1990), Cardenolide Accumulation in Crown Galls and Cell Lines Obtained by Transformation of Digitalis lanata with Agrobacterium tumefaciens Wild-Type Strains.
Cosio, E., et al J. Plant Physiol 124:155-164, Production of Antibiotic Thiarubrines by a Crown gall Tumor Line of Chaenactis douglasii.
Estramareix, C., et al. Plasmid, vol. 15 (1986) pp. 245-247.
Hanold, Dagmar, In Vitro Transformation of Protoplast-Derived Hyoscyamus Muticus Cells by Agrobacterium Tumefaciens, Plant Science Letters, 30 (1983) 177-183.
Science News, Feb. 22, 1992, p. 124.
Zambryski, P., et al. Cell, vol. 56 (1989) pp. 193-201.
Gasser, C., and Fraley, R., Transgenic Crops, Scientific American, Jun. 1992, pp. 62-69.
Komari, T., Plant Science, vol. 60 (1989) pp. 223-229.
de Cleene, M., A Possibility for Increasing the Production of Physiologically Active Substances by In Vitro Plant Tumour Cultures, Speculations in Science and Technology, vol. 3, No. 3, Aug. 1988 (Elsevier).
Esau, K, Plant Anatomy, 2nd Edition, N.Y., John Wiley & Sons, 1965, pp. 404-405.
Norton, R., Finlayson, A., and Towers, G., Thiophene Production by Crown Galls and Callus Tissues of Tagetes Patula, Phytochemistry vol. 24, No. 4, pp. 719-722, 1985.
Koziel, Michael G. et al, A Cauliflower Mosaic Virus Promoter Directs Expression of Kanamycin Resistence in Morphogenic Transformed Plant Cells. Journal of Molecular & Applied Genetics, (1984) New York; 2: 549-562.
Huizing, H., et al Acta Botanica Neerlandica, 35(1):47 (1986).

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