Claims
- 1. A method of reducing expression of a target gene in a cell, the method comprising the step of expressing in the cell an expression cassette comprising a promoter operably linked to a sense or antisense targeting sequence having substantial identity to at least a subsequence of the target gene, and an inverted repeat of a subsequence of an NOS gene, wherein the inverted repeat is heterologous to the targeting sequence, thereby reducing expression of the target gene.
- 2. The method of claim 1, wherein the inverted repeat is in a position 3′ to the targeting sequence.
- 3. The method of claim 1, wherein the inverted repeat is in a position 5′ to the targeting sequence.
- 4. The method of claim 1, wherein the inverted repeat is from the 3′ untranslated region of the NOS gene.
- 5. The method of claim 4, wherein the inverted repeat is from the terminator region of the NOS gene.
- 6. The method of claim 1, wherein the inverted repeat is from the 5′ untranslated region of the NOS gene.
- 7. The method of claim 1, wherein the inverted repeat is from the coding region of the NOS gene.
- 8. The method of claim 1, wherein the NOS gene is from an Agrobacterium sp.
- 9. The method of claim 1, wherein the inverted repeat comprises a sense region, a linker region, and an antisense region.
- 10. The method of claim 1, wherein the inverted repeat is from about 30 to about 200 nucleotides in length.
- 11. The method of claim 1, wherein the targeting sequence is a sense sequence.
- 12. The method of claim 1, wherein the targeting sequence is an antisense sequence.
- 13. The method of claim 1, wherein the targeting sequence has substantial identity to a plant pathogen target gene.
- 14. The method of claim 13, wherein the targeting sequence is a viral sequence, a bacterial sequence, an insect sequence, a fungal sequence, or a nematode sequence.
- 15. The method of claim 1, wherein the targeting sequence has substantial identity to a plant target gene.
- 16. The method of claim 1, wherein the targeting sequence is from about 100 to about 1000 nucleotides in length.
- 17. The method of claim 1 wherein the targeting sequence is from a coding region of the target gene.
- 18. The method of claim 1, wherein the targeting sequence is from a 5′ untranslated region of the target gene.
- 19. The method of claim 1, wherein the targeting sequence is from a 3′ untranslated region of the target gene.
- 20. The method of claim 1, wherein the target gene is polygalacturonase.
- 21. The method of claim 1, wherein the promoter is a tissue specific promoter.
- 22. The method of claim 1, wherein the promoter is a plant promoter.
- 23. The method of claim 22, wherein the promoter is a cauliflower mosaic virus 35S promoter or a figwort mosaic virus 34S promoter.
- 24. The method of claim 1, wherein the cell is a plant cell.
- 25. The method of claim 24, wherein the plant is selected from the group consisting of wheat, corn, rice, sorghum, pepper, tomato, squash, banana, strawberry, carrot, bean, cabbage, beet, cotton, grape, pea, pineapple, potato, soybean, yam, and alfalfa.
- 26. The method of claim 1, wherein the expression cassette has a nucleotide sequence of SEQ ID NO:1.
- 27. The method of claim 1, wherein the targeting sequence comprises a premature stop codon that inhibits translation of the targeting sequence.
- 28. An expression cassette comprising a promoter operably linked to a sense or antisense targeting sequence having substantial identity to at least a subsequence of the target gene, and an inverted repeat of a subsequence of an NOS gene, wherein the inverted repeat is heterologous to the targeting sequence.
- 29. The expression cassette of claim 28, wherein the inverted repeat is in a position 3′ to the targeting sequence.
- 30. The expression cassette of claim 28, wherein the inverted repeat is in a position 5′ to the targeting sequence.
- 31. The expression cassette of claim 28, wherein the inverted repeat is from the 3′ untranslated region of the NOS gene.
- 32. The expression cassette of claim 31, wherein the inverted repeat is from the terminator region of the NOS gene.
- 33. The expression cassette of claim 28, wherein the inverted repeat is from the 5′ untranslated region of the NOS gene.
- 34. The expression cassette of claim 28, wherein the inverted repeat is from the coding region of the NOS gene.
- 35. The expression cassette of claim 28, wherein the NOS gene is from an Agrobacterium sp,
- 36. The expression cassette of claim 28, wherein the inverted repeat comprises a sense region, a linker region, and an antisense region.
- 37. The expression cassette of claim 28, wherein the inverted repeat is from about 30 to about 200 nucleotides in length.
- 38. The expression cassette of claim 28, wherein the targeting sequence is a sense sequence.
- 39. The expression cassette of claim 28, wherein the targeting sequence is an antisense sequence.
- 40. The expression cassette of claim 28, wherein the targeting sequence has substantial identity to a plant pathogen target gene.
- 41. The expression cassette of claim 40, wherein the targeting sequence is a viral sequence, a bacterial sequence, an insect sequence, a fungal sequence, or a nematode sequence.
- 42. The expression cassette of claim 28, wherein the targeting sequence has substantial identity to a plant target gene.
- 43. The expression cassette of claim 28, wherein the targeting sequence is from about 100 to about 1000 nucleotides in length.
- 44. The expression cassette of claim 28, wherein the targeting sequence is from a coding region of the target gene.
- 45. The expression cassette of claim 28, wherein the targeting sequence is from a 5′ untranslated region of the target gene.
- 46. The expression cassette of claim 28, wherein the targeting sequence is from a 3′ untranslated region of the target gene.
- 47. The expression cassette of claim 42, wherein the target gene is polygalacturonase.
- 48. The expression cassette of claim 28, wherein the promoter is a tissue specific promoter.
- 49. The expression cassette of claim 28, wherein the promoter is a plant promoter.
- 50. The expression cassette of claim 49, wherein the promoter is a cauliflower mosaic virus 35S promoter or a figwort mosaic virus 34S promoter.
- 51. The expression cassette of claim 28, wherein the expression cassette has a nucleotide sequence of SEQ ID NO: 1.
- 52. The expression cassette of claim 28, wherein the targeting sequence comprises a premature stop codon that inhibits translation of the targeting sequence.
- 53. A transgenic plant comprising the expression cassette of claim 28.
CROSS-REFERENCES TO RELATED APPLICATIONS
[0001] The present application claims the benefit of U.S. Ser. No. 60/225,508, filed Aug. 15, 2000, herein incorporated by reference in its entirety.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60225508 |
Aug 2000 |
US |