Claims
- 1. A chimeric polypeptide comprising:
a. a first polypeptide segment that exhibits histone acetyltransferase activity; and b. a second polypeptide segment, wherein said second polypeptide segment has 40% or greater sequence identity to a subunit of a histone deacetylase chromatin-associated protein complex, wherein a terminus of said second polypeptide segment is linked to a terminus of said first polypeptide segment via at least one covalent bond.
- 2. The chimeric polypeptide of claim 1, wherein said second polypeptide has 60% or greater sequence identity to a subunit of a histone deacetylase chromatin-associated protein complex.
- 3. The chimeric polypeptide of claim 1, wherein said second polypeptide has 80% or greater sequence identity to a subunit of a histone deacetylase chromatin-associated protein complex.
- 4. The chimeric polypeptide of claim 1, wherein said second polypeptide has 90% or greater sequence identity to a subunit of a histone deacetylase chromatin-associated protein complex.
- 5. The chimeric polypeptide of claim 1, wherein said subunit exhibits scaffold activity.
- 6. The chimeric polypeptide of claim 1, wherein said subunit exhibits DNA binding activity.
- 7. The chimeric polypeptide of claim 1, wherein said subunit exhibits ATPase-dependent helicase activity.
- 8. The chimeric polypeptide of claim 1, wherein said subunit exhibits histone deacetylase activity.
- 9. The chimeric polypeptide of claim 1, wherein said first and said second polypeptide segments are directly linked via a peptide bond.
- 10. The chimeric polypeptide of claim 9, wherein the C-terminal amino acid of said first polypeptide segment is linked to the N-terminal amino acid of said second polypeptide segment.
- 11. The chimeric polypeptide of claim 9, wherein the N-terminal amino acid of said first polypeptide segment is linked to the C-terminal amino acid of said second polypeptide segment.
- 12. The chimeric polypeptide of claim 1, wherein said first and said second polypeptide segments are indirectly linked via one or more intervening amino acids that are situated between said first and said second polypeptide segments.
- 13. The chimeric polypeptide of claim 12, wherein the C-terminal amino acid of said first polypeptide segment is linked to one of said one or more intervening amino acids, and wherein the N-terminal amino acid of said second polypeptide segment is linked to one of said one or more intervening amino acids.
- 14. The chimeric polypeptide of claim 12, wherein the N-terminal amino acid of said first polypeptide segment is linked to one of said one or more intervening amino acids, and wherein the C-terminal amino acid of said second polypeptide segment is linked to one of said one or more intervening amino acids.
- 15. The chimeric polypeptide of claim 12, wherein said first and said second polypeptide segments are indirectly linked via 1 to 50 intervening amino acids.
- 16. The chimeric polypeptide of claim 15, wherein said first and said second polypeptide segments are indirectly linked via 10 to 50 intervening amino acids.
- 17. The chimeric polypeptide of claim 15, wherein said intervening amino acids comprise at least one alanine residue.
- 18. The chimeric polypeptide of claim 15, wherein said intervening amino acids comprise at least one glycine residue.
- 19. A nucleic acid construct encoding the polypeptide of claim 1.
- 20. A eukaryotic organism comprising the chimeric polypeptide of claim 1.
- 21. A eukaryotic organism comprising a nucleic acid encoding a chimeric polypeptide, said chimeric polypeptide comprising:
a. a first polypeptide segment that exhibits histone acetyltransferase activity; and b. a second polypeptide segment, wherein said second polypeptide segment has 40% or greater sequence identity to a subunit of a histone deacetylase chromatin-associated protein complex, wherein a terminus of said second polypeptide segment is covalently linked to a terminus of said first polypeptide segment.
- 22. A eukaryotic organism comprising:
a. a first nucleic acid construct comprising a first promoter and a transcription activator element operably linked to a coding sequence, said coding sequence encoding:
i) a first polypeptide segment that exhibits histone acetyltransferase activity; and ii) a second polypeptide segment, wherein said second polypeptide segment has 40% or greater sequence identity to a subunit of a histone deacetylase chromatin-associated protein complex, wherein a terminus of said second polypeptide segment is covalently linked to a terminus of said first polypeptide segment; and b. a second nucleic acid construct comprising a second promoter conferring cell type-specific transcription, said second promoter operably linked to a coding sequence for a polypeptide that binds said transcription activator element.
- 23. The eukaryotic organism of claim 22, wherein said eukaryotic organism is an animal.
- 24. The eukaryotic organism of claim 22, wherein said eukaryotic organism is a plant.
- 25. The eukaryotic organism of claim 24, wherein said plant is a monocot.
- 26. The eukaryotic organism of claim 25, wherein said monocot is selected from the group consisting of corn and rice.
- 27. The eukaryotic organism of claim 24, wherein said plant is a dicot.
- 28. The eukaryotic organism of claim 27, wherein said dicot is selected from the group consisting of soybean and rape.
- 29. The eukaryotic organism of claim 24, wherein said plant comprises an agent or mutation that alters the DNA methylation state in said plant relative to a corresponding plant that lacks said agent or mutation.
- 30. The eukaryotic organism of claim 29, wherein said DNA methylation state is decreased relative to a corresponding organism that lacks said agent or mutation.
- 31. The eukaryotic organism of claim 30, wherein said mutation is in a C5 DNA methyltransferase gene.
- 32. The eukaryotic organism of claim 30, wherein said agent affects expression of a C5 DNA methyltransferase gene.
- 33. The eukaryotic organism of claim 32, wherein said agent is an antisense nucleic acid.
- 34. A method for detecting the expression of one or more genes in a eukaryote, said method comprising:
a. isolating macromolecules from one or more specific cells in said eukaryote, said eukaryote comprising a nucleic acid construct that has a promoter operably linked to a coding sequence that encodes:
i) a first polypeptide segment that exhibits histone acetyltransferase activity; and ii) a second polypeptide segment, wherein said second polypeptide segment has 40% or greater sequence identity to a subunit of a histone deacetylase chromatin-associated protein complex, wherein a terminus of said second polypeptide segment is covalently linked to a terminus of said first polypeptide segment; and b. determining the presence or amount of at least one of said macromolecules in at least one of said specific cells.
- 35. The method of claim 34, wherein said macromolecules are polypeptides.
- 36. The method of claim 34, wherein said macromolecules are nucleic acids.
- 37. The method of claim 34, wherein said eukaryote is an animal.
- 38. The method of claim 34, wherein said eukaryote is a plant.
- 39. The method of claim 38, wherein said promoter confers cell-type specific transcription in a plant reproductive tissue.
- 40. The method of claim 39, wherein said reproductive tissue is a flower.
- 41. The method of claim 40, wherein said reproductive tissue is selected from the group consisting of: ovule, and central cell.
- 42. The method of claim 39, wherein said reproductive tissue is a seed.
- 43. The method of claim 42, wherein said reproductive tissue is selected from the group consisting of: endosperm, embryo, and zygote.
- 44. The method of claim 38, wherein said promoter confers cell-type specific transcription in a plant vegetative tissue.
- 45. A method for detecting the expression of one or more genes in a eukaryote, said method comprising:
a. isolating macromolecules from one or more specific cells in said eukaryote, said eukaryote comprising:
i) a first nucleic acid construct having a first promoter and a transcription activator element operably linked to a coding sequence, said coding sequence encoding:
a) a first polypeptide segment that exhibits histone acetyltransferase activity; and b) a second polypeptide segment, wherein said second polypeptide segment has 40% or greater sequence identity to a subunit of a histone deacetylase chromatin-associated protein complex, wherein a terminus of said second polypeptide segment is covalently linked to a terminus of said first polypeptide segment; and
ii) a second nucleic acid construct comprising a second promoter conferring cell type-specific transcription, said second promoter operably linked to a coding sequence for a polypeptide that binds said transcription activator element; and b. determining the presence or amount of at least one of said macromolecules in at least one of said specific cells.
- 46. The method of claim 45, wherein said macromolecules are polypeptides.
- 47. The method of claim 45, wherein said macromolecules are nucleic acids.
- 48. The method of claim 45, wherein said eukaryote is an animal.
- 49. The method of claim 45, wherein said eukaryote is a plant.
- 50. The method of claim 49, wherein said second promoter confers cell-type specific transcription in a plant reproductive tissue.
- 51. The method of claim 50, wherein said reproductive tissue is a flower.
- 52. The method of claim 51, wherein said reproductive tissue is selected from the group consisting of: ovule, and central cell.
- 53. The method of claim 50, wherein said reproductive tissue is a seed.
- 54. The method of claim 53, wherein said reproductive tissue is selected from the group consisting of: endosperm, embryo, and zygote.
- 55. The method of claim 49, wherein said second promoter confers cell-type specific transcription in a plant vegetative tissue.
- 56. A method for modulating gene expression in a eukaryote, said method comprising making a eukaryote having a nucleic acid construct comprising a cell-type specific promoter operably linked to a coding sequence that encodes:
a. a first polypeptide segment that exhibits histone acetyltransferase activity; and b. a second polypeptide segment, wherein said second polypeptide segment has 40% or greater sequence identity to a subunit of a histone deacetylase chromatin-associated protein complex, wherein a terminus of said second polypeptide segment is covalently linked to a terminus of said first polypeptide segment; and wherein said eukaryote exhibits modulated gene expression in cells in which said promoter confers cell-type specific transcription.
- 57. The method of claim 56, wherein said eukaryote has compositional alterations relative to a corresponding organism that lacks said nucleic acid construct.
- 58. The method of claim 56, wherein said eukaryote has developmental alterations relative to a corresponding organism that lacks said nucleic acid construct.
- 59. The method of claim 56, wherein said eukaryote has phenotypic alterations relative to a corresponding organism that lacks said nucleic acid construct.
- 60. The method of claim 56, wherein said eukaryote is an animal.
- 61. The method of claim 56, wherein said eukaryote is a plant.
- 62. The method of claim 61, wherein said promoter confers cell-type specific transcription in a plant reproductive tissue.
- 63. The method of claim 62, wherein said reproductive tissue is a flower.
- 64. The method of claim 63, wherein said reproductive tissue is selected from the group consisting of: ovule, and central cell.
- 65. The method of claim 62, wherein said reproductive tissue is a seed.
- 66. The method of claim 65, wherein said reproductive tissue is selected from the group consisting of: endosperm, embryo, and zygote.
- 67. The method of claim 61, wherein said promoter confers cell-type specific transcription in a plant vegetative tissue.
- 68. The method of claim 61, wherein said plant comprises an agent or mutation that alters the DNA methylation state in said plant relative to a corresponding plant that lacks said agent or mutation.
- 69. The method of claim 68, wherein said DNA methylation state is decreased relative to a corresponding plant that lacks said agent or mutation.
- 70. The method of claim 69, wherein said mutation is in a C5 DNA methyltransferase gene.
- 71. The method of claim 69, wherein said agent affects expression of a C5 DNA methyltransferase gene.
- 72. The method of claim 71, wherein said agent is an antisense nucleic acid.
- 73. The method of claim 61, wherein said modulated gene expression alters seed development.
- 74. The method of claim 61, wherein said modulated gene expression alters embryo development.
- 75. The method of claim 61, wherein said modulated gene expression alters endosperm development.
- 76. The method of claim 61, wherein said modulated gene expression alters seed yield by mass.
- 77. A method for modulating gene expression in a eukaryote, said method comprising making a eukaryote, said eukaryote comprising:
a. a first nucleic acid construct having a first promoter and a transcription activator element operably linked to a coding sequence, said coding sequence encoding:
i) a first polypeptide segment that exhibits histone acetyltransferase activity; and ii) a second polypeptide segment, wherein said second polypeptide segment has 40% or greater sequence identity to a subunit of a histone deacetylase chromatin-associated protein complex, wherein a terminus of said second polypeptide segment is covalently linked to a terminus of said first polypeptide segment; and b) a second nucleic acid construct comprising a second promoter conferring cell type-specific transcription, said second promoter operably linked to a coding sequence for a polypeptide that binds said transcription activator element, wherein said eukaryote exhibits modulated gene expression in cells in which said second promoter confers cell-type specific expression.
- 78. The method of claim 77, wherein said eukaryote has compositional alterations relative to a corresponding organism that lacks said nucleic acid construct.
- 79. The method of claim 77, wherein said eukaryote has developmental alterations relative to a corresponding organism that lacks said nucleic acid construct.
- 80. The method of claim 77, wherein said eukaryote has phenotypic alterations relative to a corresponding organism that lacks said nucleic acid construct.
- 81. The method of claim 77, wherein said eukaryote is an animal.
- 82. The method of claim 77, wherein said eukaryote is a plant.
- 83. The method of claim 82, wherein said promoter confers cell-type specific transcription in a plant reproductive tissue.
- 84. The method of claim 83, wherein said reproductive tissue is a flower.
- 85. The method of claim 84, wherein said reproductive tissue is selected from the group consisting of: ovule, and central cell.
- 86. The method of claim 83, wherein said reproductive tissue is a seed.
- 87. The method of claim 86, wherein said reproductive tissue is selected from the group consisting of: endosperm, embryo, and zygote.
- 88. The method of claim 82, wherein said promoter confers cell-type specific transcription in a plant vegetative tissue.
- 89. The method of claim 82, wherein said plant comprises an agent or mutation that alters the DNA methylation state in said plant relative to a corresponding plant that lacks said agent or mutation.
- 90. The method of claim 89, wherein said DNA methylation state is decreased relative to a corresponding plant that lacks said agent or mutation.
- 91. The method of claim 90, wherein said mutation is in a C5 DNA methyltransferase gene.
- 92. The method of claim 90, wherein said agent affects expression of a C5 DNA methyltransferase gene.
- 93. The method of claim 92, wherein said agent is an antisense nucleic acid.
- 94. The method of claim 82, wherein said modulated gene expression alters seed development.
- 95. The method of claim 82, wherein said modulated gene expression alters embryo development.
- 96. The method of claim 82, wherein said modulated gene expression alters endosperm development.
- 97. The method of claim 82, wherein said modulated gene expression alters seed yield by mass.
- 98. A method for making a genetically modified eukaryote, said method comprising:
a. providing a first eukaryote comprising a first nucleic acid construct, said first nucleic acid construct comprising a first promoter and a transcription activator element operably linked to a coding sequence, said coding sequence encoding:
i) a first polypeptide segment that exhibits histone acetyltransferase activity; and ii) a second polypeptide segment, wherein said second polypeptide segment has 40% or greater sequence identity to a subunit of a histone deacetylase chromatin-associated protein complex, wherein a terminus of said second polypeptide segment is covalently linked to a terminus of said first polypeptide segment; and b. providing a second eukaryote comprising a second nucleic acid construct, said second nucleic acid construct comprising a second promoter conferring embryo-specific transcription, said second promoter operably linked to a coding sequence for a polypeptide that binds said transcription activator element, c. crossing said first eukaryote and said second eukaryote to form genetically modified progeny that are sterile.
- 99. The method of claim 98, wherein said eukaryote is an animal.
- 100. The method of claim 98, wherein said eukaryote is a plant.
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority to U.S. Application Serial No. 60/300,135, filed on Jun. 22, 2001.
Provisional Applications (1)
|
Number |
Date |
Country |
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60300135 |
Jun 2001 |
US |