Claims
- 1. A recombinant nucleic acid molecule having a structure
- 2. The recombinant nucleic acid molecule of claim 1, wherein the polypeptide is other than a seed storage protein.
- 3. The recombinant nucleic acid molecule of claim 1, wherein the polypeptide is other than a seed specific protein.
- 4. The recombinant nucleic acid molecule of claim 1, wherein the signal sequence targets a linked polypeptide to a protein storage body.
- 5. A transgenic plant comprising a nucleic acid molecule according to claim 1.
- 6. Seed of a transgenic plant according to claim 5.
- 7. The plant of claim 5 wherein the plant is a stably transformed monocotyledenous plant.
- 8. The plant of claim 7, wherein the monocotyledenous plant is a cereal selected from the group of rice, barley, maize, wheat, oat, rye, sorghum, millet, or tricalate.
- 9. The plant of claim 8 wherein the plant is a barley, corn or wheat plant.
- 10. Seed of a transgenic wheat or barley plant according to claim 9.
- 11. Seed of a transgenic plant of claim 8.
- 12. A transgenic plant according to claim 5 wherein, in the nucleic acid molecule, P is a barley hordein promoter, and SS is a barley hordein signal peptide.
- 13. A transgenic plant according to claim 12 wherein the barley hordein promoter is selected from the group consisting of barley B1 and D hordein promoters and the barley hordein signal peptide is selected from the group consisting of barley B1 and D hordein signal peptides.
- 14. The seed of claim 10, wherein the polypeptide is purified from the seed at a preselected time during seed maturation at which the polypeptide is expressed at a desired concentration.
- 15. The transgenic plant of claim 5, wherein expression of the polypeptide increases the concentration of a plant metabolite.
- 16. A recombinant nucleic acid molecule having a structure
- 17. A transgenic plant comprising a nucleic acid molecule according to claim 16.
- 18. Seed of a transgenic plant according to claim 17.
- 19. The plant of claim 16 wherein the plant is a stably transformed monocotyledenous plant.
- 20. The plant of claim 19, wherein the monocotyledenous plant is a cereal selected from the group of rice, barley, maize, wheat, oat, rye, sorghum, millet, or tricalate.
- 21. The plant of claim 20 wherein the plant is a barley, corn or wheat plant.
- 22. Seed of a transgenic wheat or barley plant according to claim 21.
- 23. A transgenic barley plant according to claim 20 wherein the barley plant is a genotype selected from the group consisting of: Harrington, Morex, Crystal, Golden Promise, Stander, Moravian III, Galena, Salome, Steptoe, Klages and Baronesse.
- 24. Seed of a transgenic barley plant according to claim 22.
- 25. A transgenic wheat plant according to claim 22 wherein the wheat plant is a genotype selected from the group consisting of: Anza, Karl, Bobwhite and Yecora Rojo.
- 26. Seed of a transgenic wheat plant according to claim 25.
- 27. A method of expressing a polypeptide in seed of a monocotyledenous plant, comprising:
(a) providing a monocotyledenous plant that is stably transformed with a nucleic acid molecule according to claim 1; and (b) growing the seeds under conditions effective to produce seed and express the polypeptide in the seed.
- 28. A method of expressing a polypeptide in seed of a monocotyledenous plant, comprising:
(a) providing a monocotyledenous plant that is stably transformed with a nucleic acid molecule according to claim 15; and (b) growing the seeds under conditions effective to produce seed and express the polypeptide in the seed.
- 29. The method of claim 28 wherein the plant is a barley or a wheat plant.
- 30. A method of producing a stably transformed monocotyledenous plant expressing a selected polypeptide in seeds of the plant, comprising:
(a) placing an immature zygotic embryo of the plant on plant growth medium comprising maltose as a sugar source, an auxin at a concentration of about 0.1 mg/L to about 5 mg/L, a cytokinin at a concentration of 0 mg/L to about 5 m/L and copper at a concentration of about 0.1 μM to about 50 μM, and incubating in dim light conditions so as to form green regenerative tissue; (b) introducing a nucleic acid molecule into the tissue to produce transformed tissue, wherein the nucleic acid molecule has a structurePh-hSS-X,wherein Ph is a hordein promoter, hSS is a hordein signal sequence, and X is a nucleic acid molecule encoding the selected polypeptide, and where Ph, hSS and X are operably linked; (c) incubating the transformed tissue on the plant growth medium such that green structures are observed on the transformed material; (d) regenerating at least one transformed plant from the green structures; and (f) growing the transformed plant to produce seed.
- 31. The method of claim 30, wherein the monocotyledonous plant is a cereal selected from the group of rice, barley, maize, wheat, oat, rye, sorghum, millet, or tricalate.
- 32. The method of claim 31 wherein the plant is a barley or a wheat plant.
- 33. The method of claim 32 wherein the barley plant is a genotype selected from the group consisting of: Harrinton, Kiorex, Crystal, Golden Promise, Stander, Moravian III, Galena, Salome, Steptoe, Klages and Baronesse.
- 34. The method of claim 32 wherein the wheat plant is a genotype selected from the group consisting of: Anza, Karl, Bobwhite and Yecora Rojo.
- 35. A method of producing a stably transformed monocotyledenous plant expressing a selected polypeptide in seeds of the plant, comprising:
(a) placing an immature zygotic embryo of the plant on plant growth medium comprising maltose as a sugar source, an auxin at a concentration of about 0.1 mg/L to about 5 mg/L, a cytokinin at a concentration of 0 mg/L to about 5 mg/L and copper at a concentration of about 0.1 μM to about 50 μM, and incubating in dim light conditions so as to form green regenerative tissue; (b) introducing a nucleic acid molecule into the tissue by to produce transformed tissue, wherein the nucleic acid molecule has a structurePh-X,wherein Ph is a hordein promoter and X is a nucleic acid molecule encoding the selected polypeptide, and where Ph and X are operably linked; (c) incubating the transformed tissue on the plant growth medium such that green structures are observed on the transformed material; (d) regenerating at least one transformed plant from the green structures; and (f) growing the transformed plant to produce seed.
- 36. The method of claim 35 wherein the plant is a barley or a wheat plant.
- 37. The method of claim 36 wherein the barley plant is a genotype selected from the group consisting of: Harrington, Morex, Crystal, Stander, Moravian III, Galena, Salome, Steptoe, Klages and Baronesse.
- 38. The method of claim 36 wherein the wheat plant is a genotype selected from the group consisting of: Anza, Karl, Bobwhite and Yecora Rojo.
- 39. A method of producing a stably transformed monocotyledenous plant expressing a selected polypeptide in seeds of the plant, comprising:
(a) placing an immature zygotic embryo of the plant on plant growth medium comprising maltose as a sugar source, an auxin at a concentration of about 0.1 mg/L to about 5 mg/L, a cytokinin at a concentration of 0 mg/L to about 5 mg/L and copper at a concentration of about 0.1 μM to about 50 μM, and incubating in dim light conditions so as to form green regenerative tissue; (b) introducing a nucleic acid molecule into the tissue by to produce transformed tissue, wherein the nucleic acid molecule has a structureP-X or P-SS-X,wherein X is a nucleic acid molecule encoding a polypeptide, P is a seed maturation-specific promoter, and SS is a signal sequence that targets a linked polypeptide to an intracellular body; (c) incubating the transformed tissue on the plant growth medium such that green structures are observed on the transformed material; (d) regenerating at least one transformed plant from the green structures; and (f) growing the transformed plant to produce seed.
- 40. The method of claim 39 wherein the plant is a barley or a wheat plant.
- 41. The method of claim 39 wherein, in the nucleic acid molecule, P is a barley hordein promoter, and SS is a barley hordein signal peptide.
- 42. The method of claim 41 wherein the barley hordein promoter is selected from the group consisting of barley B1 and D hordein promoters, and the barley hordein signal peptide is selected from the group consisting of barley B1 and D hordein signal peptides.
- 43. The recombinant nucleic acid of claim 1, wherein the promoter is selected from the group of seed maturation specific promoters comprising rice glutelins, rice oryzins, rice prolamines, barley hordeins, wheat gliadins, wheat glutelins, maize zeins, maize glutelins, oat glutelins, sorghum kasirins, millet pennisetins, and rye secalins.
- 44. The recombinant nucleic acid of claim 1 wherein the signal sequence is selected from the group of a signal peptide from a monocot seed-specific gene, including a glutelin, prolamine, hordein, gliadin, glutenin, zein, albumin, globulin, ADP glucosepyrophosphorylase, starch synthase, branching enzyme, Em, and lea.
- 45. The recombinant nucleic acid of claim 1, wherein the signal sequence is selected from signal sequences that promote secretion of polypeptides from aleurone cells during seed germination.
- 46. The recombinant nucleic acid of claim 45, wherein the signal sequence is selected from a signal sequences associated with α-amylase, protease, carboxypeptidase, endoprotease, ribonuclease, Dnase/Rnase, (1-3)-β-glucanase, (1-3)(1-4) β-glucanase, esterase, acid phosphatase, pentosamine, endoxylasase, β-xylopyranossidase, arabinofuranosidase, α-glucosidase, (1-6) α-glucanase, peroxidase or lusophospholipase.
CROSS REFERENCE TO RELATED CASE
[0001] This application claims the benefit of co-pending U.S. provisional patent application No. 60/060,510, filed Sep. 30, 1997, which is incorporated herein by reference.
ACKNOWLEDGMENT OF GOVERNMENT SUPPORT
[0002] This invention was made with Government support under Grant CA-B-PLB-5752-14 from the U.S. Department of Agriculture. The Government has certain rights to this invention.
Provisional Applications (1)
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Number |
Date |
Country |
|
60060510 |
Sep 1997 |
US |
Continuations (1)
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Number |
Date |
Country |
Parent |
09164210 |
Sep 1998 |
US |
Child |
10461634 |
Jun 2003 |
US |