Appendix 1 & 2

Opine Promoters

1. Promoters and UAS from opine synthase genes

Patents granted to Biotechnology Research And Development Corporation & Purdue Research Foundation

Actual granted independent claims

US 5955646
Claim 1 A cassette for inducible expression of a foreign gene comprising said foreign gene operably linked to a regulatory region comprising:a) a promoter derived from a mannopine synthase gene of Agrobacterium tumefaciens, b) an upstream activating sequence derived from a mannopine synthase gene of Agrobacterium tumefaciens, and c) at least one upstream activating sequence derived from an octopine synthase gene of Agrobacterium tumefaciens.
Claim 2 A method for nematode inducible expression of a foreign gene in a plant, comprising:a) linking said foreign gene to a regulatory region comprising a promoter derived from a mannopine synthase gene of Agrobacterium tumefaciens comprising 138 bases upstream of the transcription initiation site, and an upstream activating sequence derived from a mannopine synthase gene of Agrobacterium tumefaciens; b) inserting said foreign gene and said regulatory region in said plant, wherein expression is induced by nematode attack on the plant.
Claim 3 A method for nematode inducible expression of a foreign gene in a plant, comprising:a) linking said foreign gene to a regulatory region comprising:i) a promoter derived from a mannopine synthase gene of Agrobacterium tumefaciens, ii) an upstream activating sequence derived from a mannopine synthase gene of Agrobacterium tumefaciens, and iii) at least one upstream activating sequence derived from an octopine synthase gene of Agrobacterium tumefaciens;b) inserting said foreign gene and said regulatory region in said plant, wherein expression is induced by nematode attack on the plant.
Claim 4 A chimeric regulatory region for expressing genes in plants comprising at least three upstream activating sequences derived from an Agrobacterium tumefaciens octopine synthase gene operably linked to a promoter derived from an Agrobacterium tumefaciens mannopine synthase gene.
Claim 5 A cassette for expressing a foreign gene comprising the foreign gene operably linked to a chimeric regulatory region comprising at least three upstream activating sequences derived from Agrobacterium tumefaciens octopine synthase genes operably linked to a promoter derived from an Agrobacterium tumefaciens mannopine synthase gene.
Claim 6 A plasmid comprising a cassette comprising a foreign gene operably linked to a chimeric regulatory region comprising at least three upstream activating sequences derived from Agrobacterium tumefaciens octopine synthase genes operably linked to a promoter derived from an Agrobacterium tumefaciens mannopine synthase gene.
Claim 7 A method of expressing a foreign gene in a plant, comprising:a) linking said foreign gene to a chimeric regulatory region comprising at least three upstream activating sequences derived from an Agrobacterium tumefaciens octopine synthase gene operably linked to a promoter derived from an Agrobacterium tumefaciens mannopine synthase gene; and b) inserting said foreign gene and said chimeric regulatory region into a plant, wherein said plant expresses said foreign gene.
Claim 8 A transgenic plant comprising a cassette comprising a foreign gene operably linked to a chimeric regulatory region comprising at least three upstream activating sequences derived from Agrobacterium tumefaciens octopine synthase genes operably linked to a promoter derived from an Agrobacterium tumefaciens mannopine synthase gene.
Claim 9 A chimeric regulatory region for expressing genes in plants comprising at least three upstream activating sequences derived from an Agrobacterium tumefaciens octopine synthase gene operably linked to an upstream activat ing sequence derived from an Agrobacterium tumefaciens mannopine synthase gene that is operably linked to a promoter derived from an Agrobacterium tumefaciens mannopine synthase gene.
Claim 10 A cassette for expressing a foreign gene comprising the foreign gene operably linked to a chimeric regulatory region comprising at least three upstream activating sequences derived from Agrobacterium tumefaciens octopine synthase genes operably linked to an upstream activating sequence derived from an Agrobacterium tumefaciens mannopine synthase gene that is operably linked to a promoter derived from an Agrobacterium tumefaciens mannopine synthase gene.
Claim 11 A method of expressing a foreign gene in a plant, comprising:a) linking said foreign gene to a chimeric regulatory region comprising at least three upstream activating sequences derived from an Agrobacterium tumefaciens octopine synthase gene operably linked to an upstream activating sequence derived from an Agrobacterium tumefaciens mannopine synthase gene that is operably linked to a promoter derived from an Agrobacterium tumefaciens mannopine synthase gene; and b) inserting said foreign gene and said chimeric regulatory region into a plant, wherein said plant expresses said foreign gene.
Claim 12 A transgenic plant comprising a chimeric regulatory region for expressing genes in plants comprising at least three upstream activating sequences derived from an Agrobacterium tumefaciens octopine synthase gene operably linked to an upstream activating sequence derived from an Agrobacterium tumefaciens mannopine synthase gene that is operably linked to a promoter derived from an Agrobacterium tumefaciens mannopine synthase gene.

AU 687961 B
Claim 1 A chimeric regulatory region for expressing genes in plants comprising an upstream activating sequences derived from an Agrobacterium tumefaciens octopine synthase gene operably linked to a promoter derived from an Agrobacterium tumefaciens mannopine synthase gene.
Claim 2 A cassette for expressing a gene comprising a gene operably linked to a chimeric regulatory region comprising an upstream activating sequences derived from an Agrobacterium tumefaciens octopine synthase gene operably linked to a promoter derived from an Agrobacterium tumefaciens mannopine synthase gene.
Claim 4 A chimeric regulatory region for expressing genes in plants comprising at least two upstream activating sequences derived from an Agrobacterium tumefaciens opine synthase gene operably linked to a promoter derived from an Agrobacterium tumefaciens opine synthase gene, wherein at least one of said upstream activating elements are derived from a different opine synthase gene than said promoter.
Claim 8 A cassette for expressing a gene comprising a gene operably linked to a chimeric regulatory region comprising at least two upstream activating sequences derived from an Agrobacterium tumefaciens opine synthase gene operably linked to a promoter derived from an Agrobacterium tumefaciens opine synthase gene, wherein at least one of said upstream activating elements are derived from a different opine synthase gene than said promoter.
Claim 13 A cassette for inducible expression of a foreign gene comprising said foreign gene operably linked to a regulatory region comprising:a) a promoter derived from a mannopine synthase gene of Agrobacterium tumefaciens by deletion to nucleotide position -138, and b) an upstream activating sequence derived from a mannopine synthase gene of Agrobacterium tumefaciens.
Claim 15 A cassette for inducible expression of a foreign gene comprising said foreign gene operably linked to a regulatory region comprising:a) a promoter derived from a mannopine synthase gene of Agrobacterium tumefaciens, b) an upstream activating sequence derived from a mannopine synthase gene of Agrobacterium tumefaciens, and c) an upstream activating sequence derived from an octopine synthase gene of Agrobacterium tumefaciens.
Claim 16 A method for expressing a gene in a plant, comprising the steps of:a) linking said gene to a chimeric regulatory region comprising an upstream activating sequence derived from an Agrobacterium tumefaciens octopine synthase gene operably linked to a promoter derived from an Agrobacterium tumefaciens mannopine synthase gene; b) inserting said gene and said chimeric regulatory region into a plant; and c) allowing said plant to express said gene.
Claim 17 A method for expressing a gene in a plant, comprising the steps of:a) linking said gene to a chimeric regulatory region comprising at least two upstream activating sequences derived from an Agrobacterium tumefaciens opine synthase gene operably linked to a promoter derived from an Agrobacterium tumefaciens opine synthase gene, wherein at least one of said upstream activating elements are derived from a different opine synthase gene than said promoter; b) inserting said gene and said chimeric regulatory region into a plant; and c) allowing said plant to express said gene.
Claim 21 A method of inducible expression of a foreign gene in a plant, comprising:a) linking said foreign gene to a regulatory region comprising:i. a promoter derived from a mannopine synthase gene of Agrobacterium tumefaciens by deletion to nucleotide position -138, andii. an upstream activating sequence derived from a mannopine synthase gene of Agrobacterium tumefaciens;b) inserting said foreign gene and said regulatory region in said plant; and c) inducing expression of said foreign gene.
Claim 23 A method for inducible expression of a foreign gene in a plant, comprising:a) linking said foreign gene to a regulatory region comprising:i. a promoter derived from a mannopine synthase gene of Agrobacterium tumefaciens,ii. an upstream activating sequence derived from a mannopine synthase gene of Agrobacterium tumefaciens; andiii. an upstream activating sequence derived from an octopine synthase gene of Agrobacterium tumefaciens; b) inserting said foreign gene and said regulatory region in said plant; and c) inducing expression of said foreign gene.
Claim 32 A chimeric regulatory region for expressing genes in plants comprising at least three upstream activating sequences derived from an Agrobacterium tumefaciens octopine synthase gene operably linked to a promoter derived from an Agrobacterium tumefaciens mannopine synthase gene.
Claim 33 A cassette for expressing a gene comprising a gene operably linked to a chimeric regulatory region comprising at least three upstream activating sequences derived from Agrobacterium tumefaciens octopine synthase genes operably linked to a promoter derived from an Agrobacterium tumefacie ns mannopine synthase gene.
Claim 34 A method of expressing a gene in a plant, comprising the steps of:a) linking said gene to a chimeric regulatory region comprising at least three upstream activating sequences derived from an Agrobacterium tumefaciens octopine synthase gene operably linked to a promoter derived from an Agrobacterium tumefaciens mannopine synthase; b) inserting said gene and said chimeric regulatory region into a plant; and c) allowing said plant to express said gene.
Claim 37 A chimeric regulatory region for expressing a gene in a plant comprising at least three upstream activating sequences derived from an Agrobacterium tumefaciens opine synthase gene operably linked to a promoter derived from an Agrobacterium tumefaciens opine synthase gene, wherein at least one of said upstream activating elements are derived from a different opine synthase gene than said promoter.
Claim 39 A cassette for expressing a gene in a plant comprising a gene operably linked to a chimeric regulatory region comprising at least three upstream activating sequences derived from Agrobacterium tumefaciens opine synthase genes operably linked to a promoter derived from an Agrobacterium tumefaciens opine synthase gene, wherein at least one of said upstream activating elements are derived from a different opine synthase gene than said promoter.
Claim 40 A chimeric regulatory region for expressing a gene in a plant comprising at lea st three upstream activating sequences derived from an Agrobacterium tumefaciens opine synthase gene operably linked to a promoter derived from an Agrobacterium tumefaciens opine synthase gene.
Claim 41 A chimeric regulatory region for expressing a gene in a plant comprising at least two upstream activating sequences derived from an Agrobacterium tumefaciens opine synthase gene operably linked to a promoter derived from an Agrobacterium tumefaciens opine synthase gene.

Patent applications filed by Biotechnology Research and Development Corporation & Purdue Research Foundation

Actual pending independent claims

EP 729514 A1
Claim 1 A chimeric regulatory region for expressing genes in plants comprising an upst ream activating sequence derived from a first Agrobacterium tumefaciens opine synthase gene operably linked to a promoter derived from a second Agrobacterium tumefaciens opine synthase gene that is different from said first Agrobacterium tumefaciens opine synthase gene.
Claim 4 A cassette for expressing a gene comprising a gene operably linked to chimeric regulatory region comprising an upstream activating sequence derived from a first Agrobacterium tumefaciens opine synthase gene operably linked to a promoter derived from a second Agrobacterium tumefaciens opine synthase gene that is different from said first Agrobacterium tumefaciens opine synthase gene.
Claim 8 A chimeric regulatory region for expressing genes in plants comprising at least two upstream activating sequences derived from an Agrobacterium tumefaciens opine synthase gene operably linked to a promoter derived from an Agrobacterium tumefaciens opine synthase gene.
Claim 12 A cassette for expressing a gene comprising a gene operably linked to a chimeric regulatory region comprising at least two upstream activating sequences derived from Agrobacterium tumefaciens opine synthase genes operably linked to a promoter derived from a Agrobacterium tumefaciens opine synthase gene.
Claim 17 A cassette for inducible expression of a foreign gene comprising said foreign gene operably linked to a regulatory region comprising a promoter derived from a mannopine synthase gene of Agrobacterium tumefaciens by deletion to nucleotide position-138 and an upstream activating sequence derived from a mannopine synthase gene of Agrobacterium tumefaciens.
Claim 19 A cassette for inducible expression of a foreign gene comprising said foreign gene operably linked to a regulatory region comprising a promoter derived from a mannopine synthase gene of Agrobacterium tumefaciens, an upstream activating sequence derived from a mannopine synthase gene of Agrobacterium tumefaciens, and an upstream activating sequence derived from an octopine synthase gene of Agrobacterium tumefaciens.
Claim 20 A method of expressing a gene in a plant, comprising the steps of: linking said gene to a chimeric regulatory region comprising an upstream activating sequence derived from a first Agrobacterium tumefaciens opine synthase gene operably linked to a promoter derived from a second Agrobacterium tumefaciens opine synthase gene that is different from said first Agrobacterium tumefaciens opine synthase gene; inserting said gene and said chimeric regulatory region into a plant; and allowing said plant to express said gene.
Claim 23 A method of expressing a gene in a plant, comprising the steps of: linking said gene to a chimeric regulatory region comprising at least two upstream activating sequences derived from an Agrobacterium tumefaciens opine synthase gene operably linked to a promoter derived from an Agrobacterium tumefaciens opine synthase; inserting said gene and said chimeric regulatory region into a plant; and allowing said plant to express said gene.
Claim 27 A method of inducible expressing a foreign gene in a plant, comprising: linking said foreign gene to a regulatory region comprising a promoter derived from a mannopine synthase gene of Agrobacterium tumefaciens by deletion to nucleotide position-138 and an upstream activating sequence derived from a mannopine synthase gene of Agrobacterium tumefaciens; inserting said foreign gene and said regulatory region in said plant; and inducing expression of said foreign gene.
Claim 29 A method for inducible expression of a foreign gene in a plant, comprising: linking said foreign gene to a regulatory region comprising a promoter derived from a mannopine synthase gene of Agrobacterium tumefaciens, an upstream activating sequence derived from a mannopine synthase gene of Agrobacterium tumefaciens, and an upstream activating sequence derived from an octopine synthase gene of Agrobacterium tumefaciens; inserting said foreign gene and said regulatory region in said plant; and inducing expression of said foreign gene.

CA 2174954
Claim 1 A chimeric regulatory region for expressing genes in plants comprising an upstream activating sequence derived from an Agrobacterium tumefaciens octopine synthase gene operably linked to a promoter derived from an Agrobacterium tumefaciens mannopine synthase gene.
Claim 2 A cassette for expressing a gene comprising a gene operably linked to chimeric regulatory region comprising an upstream activating sequence derived from a Agrobacterium tumefaciens octopine synthase gene operably linked to a promoter derived from a Agrobacterium tumefaciens mannopine synthase gene.
Claim 4 A chimeric regulatory region for expressing genes in plants comprising at least two upstream activating sequences derived from an Agrobacterium tumefaciens opine synthase gene operably linked to a promoter derived from an Agrobacterium tumefaciens opine synthase gene, wherein at least one of said upstream activating elements are derived from a different opine synthase gene than said promoter.
Claim 8 A cassette for expressing a gene comprising a gene operably linked to a chimeric regulatory region comprising at least two upstream activating sequences derived from Agrobacterium tumefaciens opine synthase genes operably linked to a promoter derived from a Agrobacterium tumefaciens opine synthase gene, wherein at least one of said upstream activating elements are derived from a different opine synthase gene than said promoter.
Claim 13 A cassette for inducible expression of a foreign gene comprising said foreign gene operably linked to a regulatory region comprising a promoter derived from a mannopine synthase gene of Agrobacterium tumefaciens by deletion to nucleotide position -138 and an upstream activating sequence derived from a mannopine synthase gene of Agrobacterium tumefaciens.
Claim 15 A cassette for inducible expression of a foreign gene comprising said foreign gene operably linked to a regulatory region comprising a promoter derived from a mannopine synthase gene of Agrobacterium tumefaciens, an upstream activating sequence derived from a mannopine synthase gene of Agrobacterium tumefaciens, and an upstream activating sequence derived from an octopine synthase gene of Agrobacterium tumefaciens.
Claim 16 A method of expressing a gene in a plant, comprising the steps of: linking said gene to a chimeric regulatory region comprising an upstream activating sequence derived from a Agrobacterium tumefaciens octopine synthase gene operably linked to a promoter derived from a Agrobacterium tumefaciens mannopine synthase gene; inserting said gene and said chimeric regulatory region into a plant; and allowing said plant to express said gene.
Claim 17 A method of expressing a gene in a plant, comprising the steps of: linking said gene to a chimeric regulatory region comprising at least two upstream activating sequences derived from an Agrobacterium tumefaciens opine synthase gene operably linked to a promoter derived from an Agrobacterium tumefaciens opine synthase, wherein at least one of said upstream activating elements are derived from a different opine synthase gene than said promoter; inserting said gene and said chimeric regulatory region into a plant; and allowing said plant to express said gene.
Claim 21 A method of inducible expressing a foreign gene in a plant, comprising: linking said foreign gene to a regulatory region comprising a promoter derived from a mannopine synthase gene of Agrobacterium tumefaciens by deletion to nucleotide position -138 and an upstream activating sequence derived from a mannopine synthase gene of Agrobacterium tumefaciens; inserting said foreign gene and said regulatory region in said plant; and inducing expression of said foreign gene.
Claim 23 A method for inducible expression of a foreign gene in a plant, comprising: linking said foreign gene to a regulatory region comprising a promoter derived from a mannopine synthase gene of Agrobacterium tumefaciens, an upstream activating sequence derived from a mannopine synthase gene of Agrobacterium tumefaciens, and an upstream activating sequence derived from an octopine synthase gene of Agrobacterium tumefaciens; inserting said foreign gene and said regulatory region in said plant; and inducing expression of said foreign gene.
Claim 32 A chimeric regulatory region for expressing genes in plants comprising at least three upstream activating sequences derived from an Agrobacterium tumefaciens octopine synthase gene operably linked to a promoter derived from an Agrobacterium tumefaciens mannopine synthase gene.
Claim 33 A cassette for expressing a gene comprising a gene operably linked to a chimeric regulatory region comprising at least three upstream activating sequences derived from Agrobacterium tumefaciens octopine synthase genes operably linked to a promoter derived from a Agrobacterium tumefaciens mannopine synthase gene.
Claim 34 A method of expressing a gene in a plant, comprising the steps of: linking said gene to a chimeric regulatory region comprising at least three upstream activating sequences derived from an Agrobacterium tumefaciens octopinesynthase gene operably linked to a promoter derived from an Agrobacterium tumefaciens mannopine synthase; inserting said gene and said chimeric regulatory region into a plant; and allowing said plant to express said gene.

The CaMV promoters: 35S and 19S

Actual granted independent claims

Patents granted to Monsanto

US 5352605
Claim 1 A chimeric gene which is expressed in plant cells comprising:(i) a promoter from a cauliflower mosaic virus, said promoter selected from the group consisting of a CaMV (35S) promoter isolated from CaMV protein-encoding DNA sequences and a CaMV (19S) promoter isolated from CaMV protein-encoding DNA sequences, and (ii) a structural sequence which is heterologous with respect to the promoter.
Claim 4 A plant cell which comprises a chimeric gene that contains:(i) a promoter from cauliflower mosaic virus, said promoter selected from the group consisting of a CaMV (35S) promoter and a CaMV (19S) promoter, wherein said promoter is isolated from CaMV protein-encoding DNA sequences, and (ii) a structural sequence which is heterologous with respect to the promoter.
Claim 7 An intermediate plant transformation plasmid which comprises:(i) a region of homology to an Agrobacterium tumefaciens vector, (ii) a T-DNA border region from Agrobacterium tumefaciens and (iii) a chimeric gene, wherein the chimeric gene is located between the T-DNA border and the region of homology,said chimeric gene comprising:(i) a promoter from cauliflower mosaic virus, said promoter selected from the group consisting of a CaMV(35S) promoter and a CaMV(19S) promoter, and (ii) a structural sequence which is heterologous with respect to the promoter.
Claim 8 A plant transformation vector which comprises:(i) a disarmed plant tumor inducing plasmid of Agrobacterium tume faciens and (ii) a chimeric gene, wherein the chimeric gene contains a promoter from cauliflower mosaic virus, said promoter selected from the group consisting of a CaMV(35S) promoter and a CaMV(19S) promoter, and (iii) a structural sequence which is heterologous with respect to the promoter.
Claim 13 A DNA construct comprising:(i) a CaMV promoter selected from the group consisting of(a) a CaMV 35S promoter isolated from CaMV protein-encoding DNA sequences and (b) a CaMV 19S promoter isolated from CaMV protein-encoding DNA sequences, and(ii) a DNA sequence of interest heterologous to (i), wherein (ii) is under the regulatory control of (i) when said construct is transcribed in a plant cell.
Claim 14 A chimeric gene which is transcribed and translated in plant cells, said chimeric gene comprising:(i) a promoter from cauliflower mosaic virus, said promoter selected from the group consisting of:(a) a CaMV 35S promoter region free of CaMV protein-encoding DNA sequences and (b) a CaMV 19S promoter region free of CaMV protein-encoding DNA sequences, and(ii) a DNA sequence which is heterologous with respect to the promoter.
Claim 15 A chimeric gene which is expressed in plants cells comprising:(i) a promoter from a cauliflower mosaic virus, said promoter selected from the group consisting of(a) a CaMV(35S) promoter region free of CaMV protein-encoding DNA sequences and (b) a CaMV(19S) promoter region free of CaMV protein-encoding DNA sequences, and(ii) a DNA sequence which is heterologous with respect to the promoter.
Claim 16 A chimeric gene which is transcribed in plants cells comprising:(i) a promoter from a cauliflower mosaic virus, said promoter selected from the group consisting of(a) a CaMV(35S) promoter free of CaMV protein-encoding DNA sequences and (b) a CaMV(19S) promoter free of CaMV protein-encoding DNA sequences,(ii) a DNA sequence which is heterologous with respect to the promoter and (iii) a 3′ non-translated polyadenylation signal sequence.
Claim 17 A plant cell which comprises a chimeric gene where said chimeric gene comprises:(i) a promoter from cauliflower mosaic virus, said promoter selected from the group consisting of a CaMV(35S) promoter and a CaMV(19S) promoter, wherein said promoter is free of CaMV protein-encoding DNA sequences, and (ii) a DNA sequence which is heterologous with respect to the promoter and (iii) a 3′ non-translated polyadenylation signal sequence.

Patents on entire 35S and 19S promoters granted to Monsanto

Actual granted independent claims

US 5530196
Claim 1 A differentiated dicotyledonous plant comprising plant cells containing a chime ric gene which comprises:a) a promoter from cauliflower mosaic virus, said promoter selected from the group consisting of a CaMV(35S) promoter free of CaMV protein-encoding DNA sequences and a CaMV(19S) promoter free of protein-encoding DNA sequences, and b) a structural sequence which is heterologous with respect to the promoter.
Claim 4 A differentiated dicotyledonous plant comprising plant cells containing in the plant genome a chimeric gene which comprises: a promoter from cauliflower mosaic virus, said promoter selected from the group consisting of a CaMV(35S) promoter and a CaMV(19S) promoter, and a DNA sequence which is heterologous with respect to the promoter.
Claim 5 A differentiated dicotyledonous plant regenerated from plant cells, said plant cells containing a chimeric gene which comprises:i. a promoter from cauliflower mosaic virus, said promoter selected from the group consisting of a CaMV(35S) promoter and a CaMV(19S) promoter, andii. a DNA sequence which is heterologous with respect to the promoter.

US 5530196

Claim 1
A differentiated dicotyledonous plant comprising plant cells containing a chime ric gene which comprises:a) a promoter from cauliflower mosaic virus, said promoter selected from the group consisting of

1. a CaMV(35S) promoter free of CaMV protein-encoding DNA sequences and
2. a CaMV(19S) promoter free of protein-encoding DNA sequences, and

b) a structural sequence which is heterologous with respect to the promoter.

Claim 4
A differentiated dicotyledonous plant comprising plant cells containing in the plant genome a chimeric gene which comprises:

1. a promoter from cauliflower mosaic virus, said promoter selected from the group consisting of a CaMV(35S) promoter and a CaMV(19S) promoter, and
2. a DNA sequence which is heterologous with respect to the promoter.

Claim 5
A differentiated dicotyledonous plant regenerated from plant cells, said plant cells containing a chimeric gene which comprises:i. a promoter from cauliflower mosaic virus, said promoter selected from the group consisting of a CaMV(35S) promoter and a CaMV(19S) promoter, andii. a DNA sequence which is heterologous with respect to the promoter.

US 5858742
Claim 1 A method for transforming a plant cell which comprises transforming a plant cell with a chimeric DNA construct containing:a) a promoter isolated from cauliflower mosaic virus (CaMV), said promoter selected from the group consisting of a CaMV(19S) promoter derived from the CaMV(19S) gene and a CaMV(35S) promoter derived from the CaMV(35S) gene, and b) a DNA sequence which is heterologous with respect to the promoter; wherein the promoter regulates the transcription of the DNA sequence.

US 5858742

Claim 1
A method for transforming a plant cell which comprises transforming a plant cell with a chimeric DNA construct containing:a) a promoter isolated from cauliflower mosaic virus (CaMV), said promoter selected from the group consisting of

1. 1. a CaMV(19S) promoter derived from the CaMV(19S) gene and
  2. a CaMV(35S) promoter derived from the CaMV(35S) gene, and

b) a DNA sequence which is heterologous with respect to the promoter;
wherein the promoter regulates the transcription of the DNA sequence.

US 6255560
Claim 1 A chimeric gene which is expressed in plant cells comprising:a) a promoter from cauliflower mosaic virus (CaMV), wherein said promoter is the CaMV(19S) promoter or the CaMV(35S) promoter, operably linked to b) a DNA sequence which is heterologous with respect to the promoter, wherein: the promoter regulates the transcription of the DNA sequence, and the DNA sequence encodes a polypeptide conferring increased antibiotic resistance to a plant or plant cell containing the DNA sequence relative to a wild-type plant or plant cell.
Claim 3 A plant cell comprising a chimeric gene which comprises:a) a promoter from cauliflower mosaic virus (CaMV), wherein said promoter is the CaMV(19S) promoter or the CaMV(35S) promoter, operably linked to b) a DNA sequence which is heterologous with respect to the promoter, wherein: the promoter regulates the transcription of the DNA sequence, and the DNA sequence encodes a polypeptide conferring increased antibiotic resistance to the plant cell relative to a wild-type plant cell.
Claim 6 An intermediate plant transformation plasmid which comprises:a) a region of homology to an A. tumefaciens vector; b) a T-DNA border from A. tumefaciens, and c) a chimeric gene,wherein the chimeric gene is located between the T-DNA border and the region of homology, said chimeric gene comprising a promoter from cauliflower mosaic virus (CaMV), wherein said promoter is the CaMV(19S) promoter or the CaMV(35S) promoter, operably linked to a DNA sequence which is heterologous with respect to the promoter, wherein:a) the promoter regulates the transcription of the DNA sequence, and b) the DNA sequence encodes a polypeptide conferring increased antibiotic resistance to a plant or plant cell containing the DNA sequence relative to a wild-type plant or plant cell.
Claim 9 A plant transformation vector which comprises a modified plant tumor inducing plasmid of A. tumefaciens which is capable of inserting a chimeric gene into susceptible plant cells, wherein the chimeric gene comprises a promoter from cauliflower mosaic virus (CaMV), wherein said promoter is the CaMV(19S) pr omoter or the CaMV(35S) promoter, operably linked to a DNA sequence which is heterologous with respect to the promoter, wherein:a) the promoter regulates the transcription of the DNA sequence, and b) the DNA sequence encodes a polypeptide conferring increased antibiotic resistance to a plant or plant cell containing the DNA sequence relative to a wild-type plant or plant cell.
Claim 12 A differentiated dicotyledonous plant comprising plant cells containing a chimeric gene which comprises a promoter from cauliflower mosaic virus (CaMV), wherein said promoter is the CaMV(19S) promoter or the CaMV(35S) promoter, operably linked to a DNA sequence encoding said polypeptide which is heterologous with respect to the promoter, wherein:a) the promoter regulates the transcription of the DNA sequence, and b) the DNA sequence encodes a polypeptide conferring increased antibiotic resistance to the plant relative to a wild-type plant.

EP 131623 B2
Claim 1 A chimeric gene capable of expressing a neomycin phosphotransferase polypeptide in plant cell conferring antibiotic resistance to the plant when inserted into the plant genome, comprising in sequence:a) a promoter region from a ribulose-1,5-bis-phosphate carboxylase small subunit gene;b) a 5′ non-translated region;c)a structural coding sequence encoding neomycin phosphotransferase I oy II; andd) a 3′ non-translated region of a gene naturally expressed in plant cells, said region encoding a signal sequence for polyadnylation of mRNA; said promoter being heterologous with respect to the structural coding sequence. Claim 4 A chimeric gene capable of expressing a polypeptide in plant cells comprising in sequence: a) a full-length transcript promoter region isolated from cauliflower mosaic virus b) a 5′ non-translated region c) a structural coding sequence d) a 3′ non-translated region of a gene naturally expressed in plants, said region encoding a signal sequence for polyadenylation of mRNA, said structural coding sequence being heterologous with respect to said promoter region.

EP 131623 B2

Claim 1
A chimeric gene capable of expressing a neomycin phosphotransferase polypeptide in plant cell conferring antibiotic resistance to the plant when inserted into the plant genome, comprising in sequence:a) a promoter region from a ribulose-1,5-bis-phosphate carboxylase small subunit gene;b) a 5′ non-translated region;c)a structural coding sequence encoding neomycin phosphotransferase I oy II; andd) a 3′ non-translated region of a gene naturally expressed in plant cells, said region encoding a signal sequence for polyadnylation of mRNA; said promoter being heterologous with respect to the structural coding sequence.

Claim 4
A chimeric gene capable of expressing a polypeptide in plant cells comprising in sequence:

a) a full-length transcript promoter region isolated from cauliflower mosaic virus
b) a 5′ non-translated region
c) a structural coding sequence
d) a 3′ non-translated region of a gene naturally expressed in plants,
said region encoding a signal sequence for polyadenylation of mRNA,
said structural coding sequence being heterologous with respect to said promoter region.

Patents granted to the Rockefeller University

Actual granted independent claims

US 5097025
Claim 1 An isolated DNA segment consisting of the nucleotide sequence: 5′-CGACCAGCAT CGTGGAAAAA GAAGACGTTC CAACCACGTC TTCAAAGC-3’.sequence of subdomain B2 of 35S CaMV promoter.
Claim 2 A DNA sequence consisting of: a) a first nucleotide sequence 5′-CGAGGAGCAT CGTGGAAAAA GAAGACGTTC CAACCACGTC TTCAAAGC-3’* and b) a second nucleotide sequence corresponding to domain A of the CaMV 35S promoter coupled to said first nucleotide sequence by means of a synthetic multilinker.*sequence of subdomain B2 of 35S CaMV promoter.
Claim 3 A DNA sequence consisting of: a) a first nucleotide sequence 5′-CATCGTTGAAG ATGCCTCTGC CGACAGTGGT CCCAAAGATG GACCCCCACC CAC-3’* and b) a second nucleotide sequence corresponding to domain A of the CaMV35S promoter coupled to said first nucleotide sequence by means of a synthetic multilinker.*sequence of subdomain B3 of 35S CaMV promoter.
Claim 4 A DNA sequence consisting of: a) a first nucleotide sequence 5′-ATTCC ATTGCCC AGCTATCTGT CACTTTATTG TGAAGATAGT GGAAAAGGAA GGTGGCTCCT ACAAATGCCA TCATTGCGAT AAAGGAAAGG CC-3’* and b) a second nucleotide sequence corresponding to domain A of the CaMV 35S promoter coupled to said first nucleotide sequence by means of a synthetic multilinker.*sequence of subdomain B4 of 35S CaMV promoter.
Claim 5 A DNA sequence consisting of: a) a first nucleotide sequence 5′-TGAGACTTTT CAACAAAGGG TAATATCCGG AAACCTCCTC GGATT-3’* and b) a second nucleotide sequence corresponding to domain A of the CaMV 35S promoter coupled to said first nucleotide sequence by means of a synthetic multilinker.*sequence of subdomain B5 of 35S CaMV 35S promoter.
Claim 6 An isolated DNA segment consisting of the nucleotide sequence: 5′-CATCGTTGAAG ATGCCTCTGC CGACAGTGGT CCCAAAGATG GACCCCCACC CAC-3’.sequence of subdomain B3 of 35S CaMV promoter.
Claim 7 An isolated DNA segment consisting of the nucleotide sequence: 5′-ATTCC ATTGCCC AGCTATCTGT CACTTTATTG TGAAGATAGT GGAAAAGGAA GGTGGCTCCT ACAAATGCCA TCATTGCGAT AAAGGAAAGG CC-3’.sequence of subdomain B4 of 35S CaMV promoter.
Claim 8 An isolated DNA segment consisting of the nucleotide sequence: 5′-TGAGACTTTT CAACAAAGGG TAATATCCGG AAACCTCCTC GGATT-3’.sequence of subdomain B5 of 35S CaMV promoter.
Claim 9 A DNA sequence consisting of: a) a first nucleotide sequence 5′-CGAGGAGCAT CGTGGAAAAA GAAGACGTTC CAACCACGTC TTCAAAGC-3’* and b) a second nucleotide sequence corresponding to the minimal promoter region of the CaMV 35S promoter coupled to said first nucleotide sequence by means of a synthetic multilinker.*sequence of subdomain B2 of 35S CaMV promoter.
Claim 10 A DNA sequence consisting of: a) a first nucleotide sequence 5′-CATCGTTGAAG ATGCCTCTGC CGACAGTGGT CCCAAAGATG GACCCCCACC CAC-3’* and b) a second nucleotide sequence corresponding to the minimal promoter region of the CaMV 35S promoter coupled to said first nucleotide sequence by means of a s ynthetic multilinker.*sequence of subdomain B3 of CaMV 35S promoter
Claim 11 A DNA sequence consisting of: a) a first nucleotide sequence 5′-ATTCC ATTGCCC AGCTATCTGT CACTTTATTG TGAAGATAGT GGAAAAGGAA GGTGGCTCCT ACAAATGCCA TCATTGCGAT AAAGGAAAGG CC-3’* and b) a second nucleotide sequence corresponding to the minimal promoter region of the CaMV 35S promoter coupled to said first nucleotide sequence by means of a synthetic multilinker.*sequence of subdomain B4 of 35S CaMV promoter
Claim 12 A DNA sequence consisting of: a) a first nucleotide sequence 5′-TGAGACTTTT CAACAAAGGG TAATATCCGG AAACCTCCTC GGATT-3’* and b) a second nucleotide sequence corresponding to the minimal promoter region of the CaMV 35S promoter coupled to said first nucleotide sequence by means of a synthetic multilinker.*sequence of subdomain B5 of 35S CaMV promoter

Patent granted to the Rockefeller University

Actual granted independent claims

US 5110732
Claim 1 In a method for the expression of a chimeric plant gene, the improvement which comprises the use of a tissue-specific promoter fragment which causes tissue-specific expression in leaves, stems, cotyledons, and vascular tissue of the hypocotyl while causing detectable levels of expression in root vascular tissue when operably coupled directly to a DNA segment corresponding to the -72 to +8 promoter fragment of the Cauliflower Mosaic Virus 35S gene, said tissue-specific promoter fragment having the sequence:5′-TGAGACTTTT CAACAAAGGG TAATATCCGG AAACCTCCTC GGATTCCATT GCCCAGCTAT CTGTCACTTT ATTGTGAAGA TAGTGGAAAA GGAAGGTGGC TCCTACAAAT GCCATCATTG CGATAAAGGA AAGGCCATCG TTGAAGATGC CTCTGCCGAC AGTGGTCCCA AAGATGGACC CCCACCCCAC GAGGAGCATC GTGGAAAAAG AAGACGTTCC AACCACGTCT TCAAAGCAAG TGGATTGATG TGATA-3’**sequence corresponds to the complete domain B -from -343 to -90 nucleotides- of the 35S CaMV promoter
Claim 2 A chimeric plant gene comprising in sequence in the 5′ to 3′ direction a tissue-specific promoter fragment consisting essentially of the sequence:5′-TGAGACTTTT CAACAAAGGG TAATATCCGG AAACCTCCTC GGATTCCATT GCCCAGCTAT CTGTCACTTT ATTGTGAAGA TAGTGGAAAA GGAAGGTGGC TCCTACAAAT GCCATCATTG CGATAAAGGA AAGGCCATCG TTGAAGATGC CTCTGCCGAC AGTGGTCCCA AAGATGGACC CCCACCCCAC GAGGAGCATC GTGGAAAAAG AAGACGTTCC AACCACGTCT TCAAAGCAAG TGGATTGATG TGATA-3’,operably coupled directly to the -72 to +8 promoter fragment of the CaMV 35S gene, said -72 to +8 promoter fragment operably coupled to a structural gene.sequence corresponds to the complete domain B from -343 to -90 nucleotides of the 35S CaMV promoter.
Claim 5 A tissue-specific promoter fragment which functions in plants to cause tissue-specific expression in the leaves, stems, cotyledons and the vascular tissue of the hypocotyl and detachable levels of expression in root vascular tissue operably coupled directly to a DNA segment corresponding to the -72 to +8 promoter fragment of the Cauliflower Mosaic Virus 35S gene, said tissue-specific promoter fragment having the sequence from its 5′ to 3′ termini:5′-TGAGACTTTT CAACAAAGGG TAATATCCGG AAACCTCCTC GGATTCCATT GCCCAGCTAT CTGTCACTTT ATTGTGAAGA TAGTGGAAAA GGAAGGTGGC TCCTACAAAT GCCATCATTG CGATAAAGGA AAGGCCATCG TTGAAGATGC CTCTGCCGAC AGTGGTCCCA AAGATGGACC CCCACCCCAC GAGGAGCATC GTGGAAAAAG AAGACGTTCC AACCACGTCT TCAAAGCAAG TGGATTGATG TGATA-3’**sequence corresponds to the complete domain B -from -343 to -90 nucleotides- of the 35S CaMV promoter.

Plant Ubiquitin Promoter System

Patent application filed by Monsanto

Actual pending independent claims

EP 1210446
Claim 1 A DNA sequence comprising an ubiquitin regulatory system lacking heatshock elements.
Claim 2 A DNA sequence comprising an ubiquitin regulatory system that is not heat inducible.

Patents granted to Mycogen

Actual granted independent claims

US 5510474
Claim 1 An isolated DNA fragment, useful in effecting expression in both monocots and dicots of coding sequences placed 3′ to said fragment, wherein said DNA is approximately 2 kb in length, and said fragment further comprises, in the following order beginning with the 5′ most element and proceeding toward the 3′ terminus of said DNA fragment:a) two heat shock elements, which overlap; b) a promoter comprising a transcription start site; c) an intron of about 1 kb in length; d) and a translation start site; wherein said DNA fragment comprising said elements (i)-(iv) regulates gene expression in both dicots and monocots, and wherein DNA fragment comprises the nucleotide sequence shown from position -899 to 1092 of the maize ubiquitin sequence listed in FIG. 2.

US 5510474

Claim 1
An isolated DNA fragment, useful in effecting expression in both monocots and dicots of coding sequences placed 3′ to said fragment, wherein said DNA is approximately 2 kb in length, and said fragment further comprises, in the following order beginning with the 5′ most element and proceeding toward the 3′ terminus of said DNA fragment:a) two heat shock elements, which overlap;
b) a promoter comprising a transcription start site;
c) an intron of about 1 kb in length;
d) and a translation start site;
wherein said DNA fragment comprising said elements (i)-(iv) regulates gene expression in both dicots and monocots, and
wherein DNA fragment comprises the nucleotide sequence shown from position -899 to 1092 of the maize ubiquitin sequence listed in FIG. 2.

US 5614399
Claim 1 A method for selective heat shock induced enhancement of the constitutive expression of a structural gene in a plant cell comprising the steps of:a) transforming said plant cell with a DNA construct comprising an approximately 2 kb plant ubiquitin regulatory region operably joined to a DNA sequence of interest, wherein said plant ubiquitin regulatory region is from a plant ubiquitin gene and comprises: at least one heat shock element, a promoter, a transcription start site, and an intron; and b) selectively applying stress conditions of high temperature to said transformed plant cell thereby inducing enhancement in expression of said DNA sequence of interest.

US 5614399

Claim 1
A method for selective heat shock induced enhancement of the constitutive expression of a structural gene in a plant cell comprising the steps of:a) transforming said plant cell with a DNA construct comprising an approximately 2 kb plant ubiquitin regulatory region operably joined to a DNA sequence of interest, wherein said plant ubiquitin regulatory region is from a plant ubiquitin gene and comprises:

1. 1. at least one heat shock element,
  2. a promoter,
  3. a transcription start site, and
  4. an intron; and

b) selectively applying stress conditions of high temperature to said transformed plant cell thereby inducing enhancement in expression of said DNA sequence of interest.

US 6020190
Claim 1 A DNA construct comprising:a) a DNA sequence no larger than 2 kb, said DNA sequence comprising a plant ubiquitin regulatory system, wherein said regulatory system contains: a heat shock element, and an intron, said intron being located at 3′ to said heat shock element, and b) a plant-expressible structural gene wherein said structural gene is placed under the regulatory control of said plant ubiquitin regulatory system.

US 6020190

Claim 1
A DNA construct comprising:a) a DNA sequence no larger than 2 kb, said DNA sequence comprising a plant ubiquitin regulatory system, wherein said regulatory system contains:

1. 1. a heat shock element, and
  2. an intron, said intron being located at 3′ to said heat shock element, and

b) a plant-expressible structural gene wherein said structural gene is placed under the regulatory control of said plant ubiquitin regulatory system.

US 6054574
Claim 1 A DNA fragment of approximately 2 kb in length, said DNA fragment comprising a plant ubiquitin regulatory system, wherein said regulatory system contains:a) a promoter comprising a transcription start site, b) one or more heat shock elements positioned 5′ to said transcription start site, and c) an intron positioned 3′ to said transcription start site, wherein said regulatory system is capable of regulating constitutive and inducible gene expression in both dicots and monocots such that the level of said constitutive gene expression in monocots is about one-third that obtained in said inducible gene expression in monocots.
Claim 9 A recombinant DNA construct comprising:a) a DNA fragment of approximately 2 kb in length, said DNA fragment comprising a plant ubiquitin regulatory system, wherein said plant ubiquitin regulatory system contains: a promoter comprising a transcription start site, one or more heat shock elements positioned 5′ to said transcription start site, a translational start site, and an intron positioned 3′ to said transcription start site and 5′ to said translational start site, wherein said plant ubiquitin regulatory system is capable of regulating constitutive and inducible gene expression in both dicots and monocots such that said constitutive gene expression in monocots is at a level about one-third that obtained in said inducible gene expression in monocots, and b) a plant-expressible heterologous structural gene positioned 3′ to said plant ubiquitin regulatory system and c) a polyadenylation signal positioned 3′ to said structural gene, wherein said heterologous gene is placed under the regulatory control of said plant ubiquitin regulatory system.
Claim 18 A DNA fragment, useful in effecting expression in both monocots and dicots of coding sequences placed 3′ to said fragment, wherein said DNA is isolated or incorporated into a larger piece of DNA but in a position other than in the 5′ sequence of a plant ubiquitin gene, is approximately 2 kb in length, and said DNA fragment further comprises, in the following order beginning with the 5′ most element and proceeding toward the 3′ terminus of said DNA fragment:(a) one or more heat shock elements, which elements may or may not be overlapping; (b) a promoter comprising a transcription start site; and (c) an intron of about 1 kb in length; and wherein said DNA fragment comprising said elements (a)-(c) is capable of regulating gene expression in both dicots and monocots.wherein said DNA is isolated or incorporated into a larger piece of DNA but in a position other than in the 5′ sequence of a plant ubiquitin gene, meaning of this?? Does not regulate the plant ubiquitin gene, is it that? or is not related to the 5’sequence of a plant ubiquitin gene at all??

US 6054574

Claim 1
A DNA fragment of approximately 2 kb in length, said DNA fragment comprising a plant ubiquitin regulatory system, wherein said regulatory system contains:a) a promoter comprising a transcription start site,
b) one or more heat shock elements positioned 5′ to said transcription start site, and
c) an intron positioned 3′ to said transcription start site,
wherein said regulatory system is capable of regulating constitutive and inducible gene expression in both dicots and monocots such that the level of said constitutive gene expression in monocots is about one-third that obtained in said inducible gene expression in monocots.

Claim 9
A recombinant DNA construct comprising:a) a DNA fragment of approximately 2 kb in length, said DNA fragment comprising a plant ubiquitin regulatory system, wherein said plant ubiquitin regulatory system contains:

1. 1. a promoter comprising a transcription start site,
  2. one or more heat shock elements positioned 5′ to said transcription start site,
  3. a translational start site, and
    an intron positioned 3′ to said transcription start site and 5′ to said translational start site,
    wherein said plant ubiquitin regulatory system is capable of regulating constitutive and inducible gene expression in both dicots and monocots such that said constitutive gene expression in monocots is at a level about one-third that obtained in said inducible gene expression in monocots, and

b) a plant-expressible heterologous structural gene positioned 3′ to said plant ubiquitin regulatory system and
c) a polyadenylation signal positioned 3′ to said structural gene,
wherein said heterologous gene is placed under the regulatory control of said plant ubiquitin regulatory system.

Claim 18
A DNA fragment, useful in effecting expression in both monocots and dicots of coding sequences placed 3′ to said fragment, wherein said DNA is isolated or incorporated into a larger piece of DNA but in a position other than in the 5′ sequence of a plant ubiquitin gene, is approximately 2 kb in length, and said DNA fragment further comprises, in the following order beginning with the 5′ most element and proceeding toward the 3′ terminus of said DNA fragment:(a) one or more heat shock elements, which elements may or may not be overlapping;
(b) a promoter comprising a transcription start site; and
(c) an intron of about 1 kb in length;
and wherein said DNA fragment comprising said elements (a)-(c) is capable of regulating gene expression in both dicots and monocots.wherein said DNA is isolated or incorporated into a larger piece of DNA but in a position other than in the 5′ sequence of a plant ubiquitin gene, meaning of this?? Does not regulate the plant ubiquitin gene, is it that? or is not related to the 5’sequence of a plant ubiquitin gene at all??

CA 1339684
Claim 1 A DNA fragment approximately 2kb in length, said DNA fragment comprising a plant ubiquitin regulatory system, wherein said regulatory system contains: a) overlapping heat shock elements and b) an intron, and wherein said regulatory system is capable of regulating constitutive and inducible gene expression in both dicots and monocots.
Claim 11 A recombinant DNA construct comprising: a) a DNA fragment approximately 2kb in length, said DNA fragment comprising a plant ubiquitin regulatory system, wherein said regulatory system contains: overlapping heat shock elements and an intron; and wherein said regulatory system is capable of regulating constitutive and inducible gene expression in both dicots and monocots, and b) a plant-expressible heterologous structural gene wherein said heterologous structural gene is placed under the regulatory control of said plant ubiquitin regulatory system.
Claim 23 A method for the constitutive expression of a structural gene and the selected stress-induced enhancement in expression of said structural gene in a plant cell comprising the steps of: a) transforming said plant cell with a DNA construct comprising an approximately 2kb plant ubiquitin regulatory system, wherein is found a heat shock element and an intron, and b) a plant-expressible structural gene that is under the regulatory control of said plant regulatory system, and c) selectively applying stress conditions of high temperature to said transformed plant cell thereby inducing enhancement in expression of said structural gene.

CA 1339684

Claim 1
A DNA fragment approximately 2kb in length, said DNA fragment comprising a plant ubiquitin regulatory system, wherein said regulatory system contains:
a) overlapping heat shock elements and
b) an intron, and
wherein said regulatory system is capable of regulating constitutive and inducible gene expression in both dicots and monocots.

Claim 11
A recombinant DNA construct comprising:
a) a DNA fragment approximately 2kb in length, said DNA fragment comprising a plant ubiquitin regulatory system, wherein said regulatory system contains:

1. 1. overlapping heat shock elements and
  2. an intron; and
    wherein said regulatory system is capable of regulating constitutive and inducible gene expression in both dicots and monocots, and

b) a plant-expressible heterologous structural gene wherein said heterologous structural gene is placed under the regulatory control of said plant ubiquitin regulatory system.

Claim 23
A method for the constitutive expression of a structural gene and the selected stress-induced enhancement in expression of said structural gene in a plant cell comprising the steps of:
a) transforming said plant cell with a DNA construct comprising an approximately 2kb plant ubiquitin regulatory system, wherein is found a heat shock element and an intron, and
b) a plant-expressible structural gene that is under the regulatory control of said plant regulatory system, and
c) selectively applying stress conditions of high temperature to said transformed plant cell thereby inducing enhancement in expression of said structural gene.

EP 342926 B1
Claim 1 A DNA sequence no larger than 2 kb, said DNA sequence comprising a plant ubiquitin regulatory system, wherein said regulatory system contains:a) a heat shock element and b) an intron.
Claim 10 A DNA construct comprising:a) a DNA sequence no larger than 2 kb, said DNA sequence comprising a plant ubiquitin regulatory system, wherein said regulatory system contains: a heat shock element, and an intron; and b) a plant-expressible structural gene wherein said structural gene is placed under the regulatory control of said plant ubiquitin regulatory system.
Claim 12 A method for the constitutive expression of a structural gene and the selected stress-induced enhancement in expression of said structural gene in a plant cell comprising the steps of:a) transforming said plant cell with a DNA construct comprising:i. a plant ubiquitin regulatory system, wherein is found a heat shock element and an intron, andii. a plant-expressible structural gene that is under the regulatory control of said plant ubiquitin regulatory system, andb) selectively applying stress conditions to said transformed plant cell thereby inducing enhancement in expression of said structural gene.

EP 342926 B1

Claim 1
A DNA sequence no larger than 2 kb, said DNA sequence comprising a plant ubiquitin regulatory system, wherein said regulatory system contains:a) a heat shock element and
b) an intron.

Claim 10
A DNA construct comprising:a) a DNA sequence no larger than 2 kb, said DNA sequence comprising a plant ubiquitin regulatory system, wherein said regulatory system contains:

1. 1. a heat shock element, and
  2. an intron; and

b) a plant-expressible structural gene wherein said structural gene is placed under the regulatory control of said plant ubiquitin regulatory system.

Claim 12
A method for the constitutive expression of a structural gene and the selected stress-induced enhancement in expression of said structural gene in a plant cell comprising the steps of:a) transforming said plant cell with a DNA construct comprising:i. a plant ubiquitin regulatory system, wherein is found a heat shock element and an intron, andii. a plant-expressible structural gene that is under the regulatory control of said plant ubiquitin regulatory system, andb) selectively applying stress conditions to said transformed plant cell thereby inducing enhancement in expression of said structural gene.

Maize alcohol dehydrogenase 1 (Adh-1) promoter

1. Patents on the ARE of maize ADH and maize aldolase promoters

Actual granted independent claims

Patent granted to Lubrizol Enterprises and CSIRO

CA 1338858
Claim 1 A recombinant DNA molecule comprising: (a) an anaerobic regulatory element; (b) a plant-expressible promoter located 3′ to said anaerobic regulatory element, and (c) a plant expressible structural gene located 3′ to said plant-expressible promoter such that said structural gene is placed under the regulatory control of said promoter and said anaerobic regulatory element, wherein said structural gene is not in nature under the regulatory control of said anaerobic regulatory element.
Claim 25 A method for selective expression of a plant-expressible structural gene under anaerobic conditions in plant tissue which comprises the steps of:a) constructing a recombinant DNA molecule which comprisesi. an anaerobic regulatory control element;ii. a plant-expressible promoter located 3′ to said anaerobic regulatory element, andiii. a plant expressible structural gene located 3′ to said plant express ible promoter such that said structural gene is placed under the regulatory control of said promoter and said anaerobic regulatory element; b) transforming said plant tissue with said recombinant DNA molecule, and c) placing said transformed plant cell under anaerobic conditions so that said plant-expressible structural gene is expressed.

CA 1338858

Claim 1
A recombinant DNA molecule comprising: (a) an anaerobic regulatory element; (b) a plant-expressible promoter located 3′ to said anaerobic regulatory element, and (c) a plant expressible structural gene located 3′ to said plant-expressible promoter such that said structural gene is placed under the regulatory control of said promoter and said anaerobic regulatory element, wherein said structural gene is not in nature under the regulatory control of said anaerobic regulatory element.

Claim 25
A method for selective expression of a plant-expressible structural gene under anaerobic conditions in plant tissue which comprises the steps of:a) constructing a recombinant DNA molecule which comprisesi. an anaerobic regulatory control element;ii. a plant-expressible promoter located 3′ to said anaerobic regulatory element, andiii. a plant expressible structural gene located 3′ to said plant express ible promoter such that said structural gene is placed under the regulatory control of said promoter and said anaerobic regulatory element;

b) transforming said plant tissue with said recombinant DNA molecule, and
c) placing said transformed plant cell under anaerobic conditions so that said plant-expressible structural gene is expressed.

Patent granted to Mycogen Plant Science Inc. and CSIRO

EP 278658 B1
Claim 1 A method for selective expression of a plant-expressible structural gene under anaerobic conditions in plant tissue, which method comprises using as an anaerobic regulatory element a recombinant DNA molecule comprising a sequence selected from: 1) 5′-GCTGGTTTCT-3′ 2) 5′-CGTGGTTTGCTTGCC-3′, or a sequence having about 66% or greater homology thereto 3) 5′-CGAGCCTTTCTTCCC-3′ 4) 5′-CTGCCTCCCTGGTTTCT-3′, and 5) 5′-CTGCAGCCCCGGTTTCG-3′, or a sequence having about 66% or greater homology thereto, a plant-expressible promoter being located 3′ to said anaerobic regulatory element, and a plant-expressible structural gene being located 3′ to said plant-expressible promoter such that said structural gene is placed under the regulatory control of said promoter and said anaerobic regulatory element.

EP 278658 B1

Claim 1
A method for selective expression of a plant-expressible structural gene under anaerobic conditions in plant tissue, which method comprises using as an anaerobic regulatory element a recombinant DNA molecule comprising a sequence selected from:
1) 5′-GCTGGTTTCT-3′
2) 5′-CGTGGTTTGCTTGCC-3′, or a sequence having about 66% or greater homology thereto
3) 5′-CGAGCCTTTCTTCCC-3′
4) 5′-CTGCCTCCCTGGTTTCT-3′, and
5) 5′-CTGCAGCCCCGGTTTCG-3′, or a sequence having about 66% or greater homology thereto,
a plant-expressible promoter being located 3′ to said anaerobic regulatory element, and a plant-expressible structural gene being located 3′ to said plant-expressible promoter such that said structural gene is placed under the regulatory control of said promoter and said anaerobic regulatory element.

Patent granted to Mycogen Plant Science Inc. and Lubrizol Enterprises

US 5001060
Claim 1 A recombinant DNA molecule comprising: (a) an anaerobic regulatory element; (b) a plant-expressible promoter located 3′ to said anaerobic regulatory element, and (c) a plant-expressible structural gene located 3′ to said plant-expressible promoter such that said structural gene is placed under the regulatory control of said promoter and said anaerobic regulatory element wherein said structural gene is not in nature under the regulatory control of said anaerobic regulatory element.

US 5001060

Claim 1
A recombinant DNA molecule comprising:
(a) an anaerobic regulatory element;
(b) a plant-expressible promoter located 3′ to said anaerobic regulatory element, and
(c) a plant-expressible structural gene located 3′ to said plant-expressible promoter
such that said structural gene is placed under the regulatory control of said promoter and said anaerobic regulatory element
wherein said structural gene is not in nature under the regulatory control of said anaerobic regulatory element.

2. Patents on the recombinant promoter EMU

Patent granted to Lubrizol Genetics Inc.

AU 643521 B2
Claim 1 A recombinant promoter molecule for enhancing expression of a plant-expressible structural gene in a monocot plant cell comprising:a) a plurality of enhancer elements selected from the group consisting of the ARE and the OCS elements; b) a truncated, plant expressible promoter providing a TATA box region necessary to initiate transcription positioned 3′ to said plurality of enhancer elements; and c) a nucleotide sequence naturally found as an intron positioned between the transcription start site and the translation start site in a plant-expressible gene; whereby a plant-expressible structural gene placed 3′ to said recombinant promoter molecule is expressed in said monocot plant cell under regulatory control of said recombinant promoter molecule.

AU 643521 B2

Claim 1
A recombinant promoter molecule for enhancing expression of a plant-expressible structural gene in a monocot plant cell comprising:a) a plurality of enhancer elements selected from the group consisting of the ARE and the OCS elements;
b) a truncated, plant expressible promoter providing a TATA box region necessary to initiate transcription positioned 3′ to said plurality of enhancer elements; and
c) a nucleotide sequence naturally found as an intron positioned between the transcription start site and the translation start site in a plant-expressible gene;
whereby a plant-expressible structural gene placed 3′ to said recombinant promoter molecule is expressed in said monocot plant cell under regulatory control of said recombinant promoter molecule.

Patent granted to Mycogen Plant Science Inc. and CSIRO

EP 459643 B1
Claim 1 A recombinant promoter molecule for enhancing expression of a plant-expressible structural gene in a monocot plant cell comprising:a) a plurality of ARE enhancer elements; b) a truncated, plant expressible promoter providing a TATA box region necessary to initiate transcription positioned 3′ to said plurality of enhancer elements; and c) a nucleotide sequence naturally found as an intron positioned between the transcription start site and the translational start site in a plant-expre ssible gene; whereby a plant-expressible structural gene placed 3′ to said recombinant promoter molecule is expressed in said monocot plant cell under regulatory control of said recombinant promoter molecule.

EP 459643 B1

Claim 1
A recombinant promoter molecule for enhancing expression of a plant-expressible structural gene in a monocot plant cell comprising:a) a plurality of ARE enhancer elements;
b) a truncated, plant expressible promoter providing a TATA box region necessary to initiate transcription positioned 3′ to said plurality of enhancer elements; and
c) a nucleotide sequence naturally found as an intron positioned between the transcription start site and the translational start site in a plant-expre ssible gene;
whereby a plant-expressible structural gene placed 3′ to said recombinant promoter molecule is expressed in said monocot plant cell under regulatory control of said recombinant promoter molecule.

Patent granted to CSIRO

US 5290924
Claim 1 A recombinant promoter molecule, useful for enhancing expression of a plant-expressible structural gene in a monocot plant cell, said promoter molecule comprising:a) a plurality of enhancer elements selected from the group consisting of only ARE elements, only OCS elements, and combinations of ARE and OCS elements; b) a truncated, plant expressible promoter, providing a TATA box region and a transcription start site, said promoter selected from the group consisting of .DELTA.35S and .DELTA.ADH positioned 3′ to said plurality of enhancer elements wherein said truncated promoter excludes the presence of enhancer sequences and wherein said truncated promoter is recombined with said plurality of enhancer elements positioned 5′ to said truncated promoter; and c) a maize Adh1 intron positioned 3′ to said transcription start site whereby a plant-expressible structural gene, placed 3′ to said recombinant promoter molecule, is expressed in said monocot plant cell under regulatory control of said recombinant promoter molecule.

US 5290924

Claim 1
A recombinant promoter molecule, useful for enhancing expression of a plant-expressible structural gene in a monocot plant cell, said promoter molecule comprising:a) a plurality of enhancer elements selected from the group consisting of only ARE elements, only OCS elements, and combinations of ARE and OCS elements;
b) a truncated, plant expressible promoter, providing a TATA box region and a transcription start site, said promoter selected from the group consisting of .DELTA.35S and .DELTA.ADH positioned 3′ to said plurality of enhancer elements wherein said truncated promoter excludes the presence of enhancer sequences and wherein said truncated promoter is recombined with said plurality of enhancer elements positioned 5′ to said truncated promoter; and
c) a maize Adh1 intron positioned 3′ to said transcription start site whereby a plant-expressible structural gene, placed 3′ to said recombinant promoter molecule, is expressed in said monocot plant cell under regulatory control of said recombinant promoter molecule.

Synthetic promoters

Patents granted to Pioneer Hi-Bred International Inc.

Actual granted independent claims

US 6072050
Claim 1 A synthetic DNA promoter sequence functional in a plant cell, said promoter sequence comprising:a TATA motif, a transcription start site, and a region between said TATA motif and said start site that is at least 64% GC-rich; wherein said region is not a region between a TATA motif and a transcription start site of native maize ubiquitin promoter, and wherein said promoter sequence is set forth in SEQ ID NO:10.
Claim 2 A synthetic DNA promoter sequence functional in a plant cell, said promoter sequence comprising:a TATA motif, a transcription start site, and a region between said TATA motif and said start site that is at least 64% GC-rich; wherein said region is not a region between a TATA motif and a transcription start site of native maize ubiquitin promoter, and wherein said promoter sequence is set forth in SEQ ID NO:1.
Claim 3 An expression cassette comprisinga synthetic promoter comprising: a TATA motif, a transcription start site and a region between said TATA motif and said start site that is at least 64% GC rich, a structural gene operatively linked to said promoter, and a transcription end site polyadenylation signal; wherein said region is not a region between a TATA motif and a transcription start site of native maize ubiquitin promoter, and wherein sequence of said promoter is set forth in SEQ ID NO:1.
Claim 4 An expression cassette comprisinga synthetic promoter comprising: a TATA motif, a transcription start site and a region between said TATA motif and said start site that is at least 64% GC rich, a structural gene operatively linked to said promoter, and a transcription end site polyadenylation signal; wherein said region is not a region between a TATA motif and a transcription start site of native maize ubiquitin promoter, and wherein sequence of said promoter is set forth in SEQ ID NO:10.
Claim 5 An expression cassette comprisinga synthetic promoter comprising: a TATA motif, a transcription start site and a region between said TATA motif and said start site that is at least 64% GC rich, a structural gene operatively linked to said promoter, a transcription end site polyadenylation signal, and an upstream element operatively linked to said promoter so that transcription is enhanced; wherein said region is not a region between a TATA motif and a transcription start site of native maize ubiquitin promoter; and wherein sequence of said upstream element is set forth in SEQ ID NO:2.
Claim 7 A synthetic upstream element having a sequence set forth in SEQ ID NO:2.
Claim 8 An expression cassette comprising:a promoter sequence; a structural gene operatively linked to said promoter sequence; a polyadenylation signal; and a synthetic upstream element comprising SEQ ID NO:2 operatively linked to said promoter so that expression is enhanced.
Claim 13 An isolated nucleotide sequence comprising a DNA enhancer sequence comprising the nucleotide sequence set forth in SEQ ID No: 5.
Claim 14 A nucleotide sequence comprising a promoter construct, said construct comprising in operable linkage a core promoter sequence and a Ubi-1 UAR, wherein said Ubi-1 UAR is a maize Ubi UAR comprising the sequence set forth in SEQ ID No: 13.
Claim 15 An expression cassette comprising in operable linkage:a core promoter sequence, a Ubi UAR operably linked upstream to said core promoter to form a synthetic promoter construct, a nucleotide sequence of interest operably linked to said synthetic promoter, and a polyadenylation signal; wherein said Ubi-1 UAR comprises the sequence set forth in SEQ ID No:13.

AU 729929 B2
Claim 1 A synthetic DNA plant promoter sequence, said sequence comprising:a TATA motif, a transcription start site, and a region between said TATA motif and said start site that is at least 64% GC-rich.
Claim 3 An expression cassette comprisinga synthetic promoter comprising a TATA motif, a transcription start site and a region there between that is at least 65% GC rich, a structural gene operatively linked to said promoter, and a transcription end site polyadenylation signal.
Claim 17 A synthetic upstream element having a sequence of SEQ ID NO:2.
Claim 18 An expression cassette comprising: a promoter sequence; a structural gene operatively linked to said promoter sequence; a polyadenylation signal; and a synthetic upstream element comprising SEQ ID NO:2 operatively linked to said promoter sequence so that expression is enhanced.
Claim 23 A synthetic DNA plant promoter sequence functional in a plant cell, said sequence comprising:a TATA motif, a transcription start site, and a region between said TATA motif and said start site that is at least 64% GC-rich; wherein said promoter sequence is less than 1000 bp.

EP 1056875 A1
Claim 1 A synthetic DNA plant promoter sequence, said sequence comprising:a TATA motif ; a transcription start site; and a region between said TATA motif and said start site that is at least about 64% GC-rich.
Claim 4 An expression cassette comprising:a synthetic promoter comprising: a TATA motif ; a transcription start site and a region there between that is at least about 64% GC rich; a structural gene operatively linked to said promoter; and a transcription end site polyadenylation signal.
Claim 19 A synthetic upstream element having a sequence of SEQ ID NO: 2.
Claim 20 An expression cassette comprising:a promoter sequence; a structural gene operatively linked to said promoter sequence; a polyadenylation signal; and a synthetic upstream element homologous to SEQ ID NO: 2 operatively linked to said promoter so that expression is enhanced.
Claim 25 A DNA sequence comprising:a promoter construct, said construct comprising in operable linkage: a core synthetic promoter sequence comprising a TATA motif, a transcription start site, and a region between said TATA motif and said start site that is at least 64% GC-rich; and an upstream activating region operably linked to said core synthetic promoter.
Claim 32 An expression cassette comprising:a core synthetic promoter sequence comprising a TATA motif, a transcription start site, and a region between said TATA motif and said start site that is at least 64% GC-rich; an upstream activating region operably linked to said synthetic promoter to enhance transcription; a structural gene operably linked to said synthetic promoter; and a polyadenylation signal.
Claim 38 A method for controlling the level of expression of a transgenic nucleotide sequence in a plant cell said method comprising transforming with an expression cassette comprising a promoter having at least one Ubi-1 UAR.
Claim 40 A nucleotide sequence comprising:a promoter construct, said construct comprising in operable linkage: a core promoter sequence; and a Ubi-I UAR.
Claim 46 An expression cassette comprising in operable linkage:a core promoter sequence; a Ubi UAR operably linked to said core promoter to form a synthetic promoter construct; a nucleotide sequence of interest operably linked to said synthetic promoter; and a polyadenylation signal.

CA 2257719 AA
Claim 1 A synthetic DNA plant promoter sequence, said sequence comprising:a TATA motif; a transcription start site; and a region between said TATA motif and said start site that is at least 64% GC rich.
Claim 4 An expression cassette comprising:a synthetic promoter comprising: a TATA motif; a transcription start site and a region there between that is at least 64% GC rich; a structural gene operatively linked to said promoter; and a transcription end site polyadenylation signal.
Claim 19 A synthetic upstream element having a sequence of SEQ ID NO:2.
Claim 20 An expression cassette comprising:a promoter sequence; a structural gene operatively linked to said promoter sequence; a polyadenylation signal; and a synthetic upstream element comprising SEQ ID NO:2 operatively linked to said promoter so that expression is enhanced.
Claim 25 A synthetic DNA promoter sequence functional in plant cells, wherein said DNA sequence is synthetically created and non-naturally occurring, consisting of:a TATA motif; a transcriptional start site; and a region between said TATA motif and said start site that is at least 64% GC rich.
Claim 26 A synthetic DNA plant promoter sequence functional in plant cells, said sequence comprising:a TATA motif; a transcription start site; a region between said TATA motif and said start site that is at least 64% GC rich; wherein said promoter sequence is less than 1900 bp.

US 2001/0047092 A1 and WO 01/53476 A2
Claim 1 A plant promoter comprising at least one synthetic multimeric promoter element region having a nucleotide sequence selected from the group consisting of:a) a nucleotide sequence comprising six DRE 1 (SEQ ID NO.: 59), two ABRE1 (SEQ ID NO.: 2), three As-1 (SEQ ID NO.: 7), one GT-2 (SEQ ID NO.: 24), and two PCNA IIA (SEQ ID NO.: 45) promoter elements; b) a nucleotide sequence comprising three DRE 1 (SEQ ID NO.: 59), three ABRE1 (SEQ ID NO.: 2), one As-1 (SEQ ID NO.: 7), two GT-2 (SEQ ID NO.: 24), and two PCNA IIA (SEQ ID NO.: 45) promoter elements; c) a nucleotide sequence comprising five DRE 1 (SEQ ID NO.: 59), three ABRE1 (SEQ ID NO.: 2), two As-1 (SEQ ID NO.: 7), and five GT-2 (SEQ ID NO.: 24) promoter elements; d) a nucleotide sequence comprising four DRE 1 (SEQ ID NO.: 59), three ABRE1 (SEQ ID NO.: 2), three GT-2 (SEQ ID NO.: 24), and one PCNA IIA (SEQ ID NO.: 45) promoter elements; e) a nucleotide sequence comprising two DRE 1 (SEQ ID NO.: 59), one ABRE1 (SEQ ID NO.: 2), five As-1 (SEQ ID NO.: 7), one GT-2 (SEQ ID NO.: 24), and three PCNA IIA (SEQ ID NO.: 45) promoter elements; f) a nucleotide sequence comprising five DRE 1 (SEQ ID NO.: 59), two ABRE1 (SEQ ID NO.: 2), one As-1 (SEQ ID NO.: 7), one GT-2 (SEQ ID NO.: 24), and two PCNA IIA (SEQ ID NO.: 45) promoter elements; g) a nucleotide sequence comprising one DRE 1 (SEQ ID NO.: 59), two ABRE1 (SEQ ID NO.: 2), two As-1 (SEQ ID NO.: 7), and one GT-2 (SEQ ID NO.: 24) promoter elements; h) a nucleotide sequence comprising two DRE 1, one ABRE 1 (SEQ ID NO.: 2), three As-1 (SEQ ID NO.: 7), and one GT-2 (SEQ ID NO.: 24) promoter elements; and i) a nucleotide sequence that hybridizes under stringent conditions to any of the nucleotide sequences of a), b), c), d), e), f), g), and h).
Claim 7 A plant, or its parts, having stably incorporated into its genome a DNA construct comprising a plant promoter operably linked to a coding sequence, said plant promoter comprising at least one synthetic multimeric promoter element region (SMPER) that enhances expression of said coding sequence.
Claim 8 A plant, or its parts, having stably incorporated into its genome a DNA construct comprising a plant promoter operably linked to a coding sequences, said plant promoter comprising at least one synthetic multimeric promoter element region having a nucleotide sequence selected from the group consisting of:a) a nucleotide sequence comprising promoter elements DRE1, ABRE1, DRE1, As-1, ABRE1, DRE1, GT-2, As-1, DRE1, PCNA IIA, PCNA IIA, DRE1, As-1, and DRE1 sequentially (SEQ ID NO.: 66); b) a nucleotide sequence comprising promoter elements DRE1, DRE1, As-1, PCNA IIA, ABRE1, PCNA IIA, ABRE1, DRE1, GT-2, GT-2, and ABRE1 sequentially (SEQ ID NO.: 67); c) a nucleotide sequence comprising promoter elements GT-2, ABRE1, ABRE1, GT-2, As-1, GT-2, GT-2, DRE1, GT-2, DRE1, DRE1, As-1, DRE1, DRE1, and ABRE1 sequentially (SEQ ID NO.: 65); d) a nucleotide sequence comprising promoter elements ABRE1, ABRE1, GT-2, GT-2, GT-2, DRE1, DRE1, DRE1, DRE1, ABRE1, and PCNA IIA sequentially (SEQ ID NO.: 68); e) a nucleotide sequence comprising promoter elements PCNA IIA, As-1, GT-2, As-1, DRE1, As-1, As-1, PCNA IIA, As-1, PCNA IIA, DRE1, and ABRE1 sequentially (SEQ ID NO.: 69); f) a nucleotide sequence comprising promoter elements As-1, GT-2, DRE1, DRE1, ABRE1, PCNA IIA, DRE1, PCNA IIA, ABRE1, DRE1, and DRE1 sequentially (SEQ ID NO.: 71); g) a nucleotide sequence comprising promoter elements As-1, ABRE 1, GT-2, As-1, ABRE1, and DRE1 sequentially (SEQ ID NO.: 72); h) a nucleotide sequence comprising promoter elements DRE 1, ABRE1, GT-2, DRE1, As-1, As-1, and As-1 sequentially (SEQ ID NO.: 70); i) a nucleotide sequence set forth in FIGS. 7, 8, 9, 10, 11, 12, 13, or 14 (SEQ ID NOS.: 65-72); j) a nucleotide sequence that comprises a variant of a nucleotide sequence set forth in FIGS. 7, 8, 9, 10, 11, 12, 13, or 14 (SEQ ID NOS.: 65-72); and k) a nucleotide sequence that hybridizes under stringent conditions to a nucleotide sequence of (a), (b), (c), (d), (e), (f), (g), (h), (i), or (j).
Claim 12 A plant cell having stably incorporated into its genome a DNA construct comprising a plant promoter operably linked to a coding sequence, said plant promoter comprising at least one synthetic multimeric promoter element region having a nucleotide sequence selected from the group consisting of:a) a nucleotide sequence comprising promoter elements DRE1, ABRE1, DRE1, As-1, ABRE1, DRE1, GT-2, As-1, DRE1, PCNA IIA, PCNA IIA, DRE1, As-1, and DRE1 sequentially (SEQ ID NO.: 66); b) a nucleotide sequence comprising promoter elements DRE1, DRE1, As-1, PCNA IIA, ABRE1, PCNA IIA, ABRE1, DRE1, GT-2, GT-2, and ABRE1 sequentially (SEQ ID NO.: 67); c) a nucleotide sequence comprising promoter elements GT-2, ABRE1, ABRE1, GT-2, As-1, GT-2, GT-2, DRE1, GT-2, DRE1, DRE1, As-1, DRE1, DRE1, and ABRE1 sequentially (SEQ ID NO.: 65); d) a nucleotide sequence comprising promoter elements ABRE 1, ABRE1, GT-2, GT- 2, GT-2, DRE1, DRE1, DRE1, DRE1, ABRE1 and PCNA IIA sequentially (SEQ ID NO.: 68); e) a nucleotide sequence comprising promoter elements PCNA IIA, As-1, GT-2, As-1, DRE1, As-1, As-1, PCNA IIA, As-1, PCNA IIA, DRE1, and ABRE1 sequentially (SEQ ID NO.: 69); f) a nucleotide sequence comprising promoter elements As-1, GT-2, DRE1, DRE1, ABRE1, PCNA IIA, DRE1, PCNA IIA, ABRE1, DRE1, and DRE1 sequentially (SEQ ID NO.: 71); g) a nucleotide sequence comprising promoter elements As-1, ABRE1, GT-2, As-1, ABRE1, and DRE1 sequentially (SEQ ID NO.: 72); h) a nucleotide sequence comprising promoter elements DRE1, ABRE1, GT-2, DRE1, As-1, As-1, and As-1 sequentially (SEQ ID NO.: 70); i) a nucleotide sequence set forth in FIGS. 7, 8, 9, 10, 11, 12, 13, or 14 (SEQ ID NOS.: 65-72); j) a nucleotide sequence that comprises a variant of a nucleotide sequence set forth in FIGS. 7, 8, 9, 10, 11, 12, 13, or 14 (SEQ ID NOS.: 65-72); and k) a nucleotide sequence that hybridizes under stringent conditions to the nucleotide sequence of (a), (b), (c), (d), (e), (f), (g), (h), (i), or (j).
Claim 17 A method of selecting promoter elements active in a tissue of interest, comprising:a) isolating or synthesizing oligonucleotides representing known or putative promoter elements or transcription factor binding sites; b) labeling said oligonucleotides; c) pooling said oligonucleotides to create an array which facilitates screening; d) hybridizing said oligonucleotides with nuclear extracts of said tissue of interest; and e) selecting those oligonucleotides exhibiting preferential binding to said nuclear extracts.
Claim 18 A method of creating synthetic multimeric promoter element regions active in a tissue of interest, comprising:a) selecting known or putative promoter elements or transcription factor binding sites which exhibit preferential binding to nuclear extract prepared from said tissue of interest; b) combining said selected oligonucletides in novel arrangements encompassing variation in number of copies, sequential order, orientation, and spacer regions; and c) testing said novel arrangements for their effect on transcription and selecting those demonstrating enhancement or suppression of linked gene expression.

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Appendix 1 & 2

Opine Promoters

The CaMV promoters: 35S and 19S

Plant Ubiquitin Promoter System

Maize alcohol dehydrogenase 1 (Adh-1) promoter

Synthetic promoters