Patent EP0332104A2 successfully refreshed from Google Patents.
EP0332104A2
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EP
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Chemically regulatable DNA sequences and genes and uses thereof
Assignee: Syngenta Participations AG
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Filing: 1989-03-06
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Granted: —
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Expires: 2009-03-06
Abstract
Abstract The present invention provides chemically regulatable DNA sequences capable of regulating transcription of an associated DNA sequence in plants or plant tissues, chimeric constructions containing such sequences, vectors containing such sequences and chimeric constructions, and transgenic plants and plant tissues containing these chimeric constructions. Also provided are a novel signal peptide sequence, genes which code for this sequence, and newly identified PR protein genes. The chimeric constructions and transgenic plants and plant tissues may be used in an assay for new chemical regulators.
Inventors
- 👤 John Ryals
- 👤 Alice Montoya
- 👤 Christian Harms
- 👤 John Duesing
- 👤 Christoph Sperisen
- 👤 Fred Meins
- 👤 George Payne
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-29 14:05
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/66; C07; C07D; C07D285/00; C07D285/01; C07D285/02; C07D285/14; C07K; C07K14/00; C07K14/415; C12N; C12N15/00; C12N15/09; C12N15/11; C12N15/62; C12N15/625; C12N15/63; C12N15/79; C12N15/82; C12N15/8216; C12N15/8237; C12N15/8238; C12N15/8241; C12N15/8261; C12N15/8271; C12N15/8274; C07K2319/00; C07K2319/01; C07K2319/02; C07K2319/55; C07K2319/61; C12Q2600/00; C12Q2600/13; C12Q2600/142
Claims Analysis
Total Claims: 133
Claim 1
Independent Claim
29 words
A non-coding DNA sequence capable of regulating the transcription of an associated DNA sequence in a plant or plant tissue wherein this regulation is dependent upon a chemical regulator.
Claim 2
Dependent Claim
29 words
A non-coding DNA sequence capable of regulating the transcription of an associated DNA sequence in a plant or plant tissue wherein this regulation is dependent upon a chemical regulator.
Claim 3
Dependent Claim
10 words
A substantially pure non-coding DNA sequence according to claim 1.
Claim 4
Dependent Claim
16 words
A DNA sequence according to claim 2 wherein said associated DNA sequence is a coding sequence.
Claim 5
Dependent Claim
17 words
A DNA sequence according to claim 2 wherein said regulation is dependent upon an inducing chemical regulator.
Claim 6
Dependent Claim
33 words
A substantially pure DNA sequence according to claim 1 which is, or has substantial sequence homology to, a non-coding chemically regulatable DNA sequence which is part of a naturally occurring chemically regulatable gene.
Claim 7
Dependent Claim
32 words
A DNA sequence according to claim 5 capable of regulating the transcription of an associated DNA sequence in a plant or plant tissue wherein said associated DNA sequence is a coding sequence.
Claim 8
Dependent Claim
17 words
A DNA sequence according to claim 6 wherein said transcription is regulated by an inducing chemical regulator.
Claim 9
Dependent Claim
17 words
A DNA sequence according to claim 5 wherein said chemically regulatable gene is a PR protein gene.
Claim 10
Dependent Claim
35 words
A DNA sequence according to claim 8 wherein said PR protein gene is a tobacco PR-1a, PR-1b, PR-1c, PR-1′, PR-Q or PR-R gene, a cucumber chitinase gene, or a tobacco basic or acidic β-1,3-glucanase gene.
Claim 11
Dependent Claim
27 words
A DNA sequence according to claim 8 wherein said PR protein gene is a tobacco PR-1a, PR-1b, PR-1c, PR-1′ or PR-R gene or a cucumber chitinase gene.
Claim 12
Dependent Claim
17 words
A DNA sequence according to claim 8 wherein said PR protein gene is a tobacco PR-1a gene.
Claim 13
Dependent Claim
17 words
A DNA sequence according to claim 8 wherein said PR protein gene is a tobacco PR-1′ gene.
Claim 14
Dependent Claim
18 words
A DNA sequence according to claim 8 wherein said PR protein gene is a basic tobacco β-1,3-glucanase gene.
Claim 15
Dependent Claim
19 words
A DNA sequence according to claim 8 which is located in the 5′-flanking region of said PR protein gene.
Claim 16
Dependent Claim
33 words
A DNA sequence according to claim 6 wherein said transcription is regulated by a regulator selected from the group consisting of benzoic acid, salicylic acid, acetylsalicylic acid, polyacrylic acid and substituted derivatives thereof.
Claim 17
Dependent Claim
35 words
A DNA sequence according to claim 6 wherein said transcription is regulated by a regulator selected from the group consisting of benzo-1,2,3-thiadiazolecarboxylic acid, benzo-1,2,3-thiadiazolethiocarboxylic acid, cyanobenzo-1,2,3-thiadiazole, benzo-1,2,3-thiadiazolecarboxylic acid amide, benzo-1,2,3-thiadiazolecarboxylic acid hydrazide, and derivatives thereof.
Claim 18
Dependent Claim
15 words
A DNA sequence according to claim 16 wherein said transcription is regulated by methyl benzo-1,2,3-thiadiazole-7-carboxylate.
Claim 19
Dependent Claim
19 words
A DNA sequence according to claim 6 wherein said transcription is regulated by dichloroisonicotinic acid or a derivative thereof.
Claim 20
Dependent Claim
98 words
A DNA sequence which comprises a first DNA component sequence which is, or has substantial sequence homology to, a non-coding, chemically regulatable DNA sequence of a naturally occurring chemically regulatable gene, this first component sequence being capable of regulating the transcription of an associated DNA sequence in a plant or plant tissue, wherein this regulation is dependent upon a chemical regulator, and a second DNA component sequence which is, or has substantial sequence homology to, part but not all of a transcribable DNA sequence with which the first component is associated in the naturally occurring chemically regulatable gene.
Claim 21
Dependent Claim
98 words
A DNA sequence which comprises a first DNA component sequence which is, or has substantial sequence homology to, a non-coding, chemically regulatable DNA sequence of a naturally occurring chemically regulatable gene, this first component sequence being capable of regulating the transcription of an associated DNA sequence in a plant or plant tissue, wherein this regulation is dependent upon a chemical regulator, and a second DNA component sequence which is, or has substantial sequence homology to, part but not all of a transcribable DNA sequence with which the first component is associated in the naturally occurring chemically regulatable gene.
Claim 22
Dependent Claim
9 words
A substantially pure DNA sequence according to claim 19.
Claim 23
Dependent Claim
28 words
A DNA sequence according to claim 20 wherein said naturally occurring chemically regulatable gene occurs in a plant and said second DNA component sequence is a coding sequence.
Claim 24
Dependent Claim
15 words
A DNA sequence according to claim 21 wherein said gene is a PR protein gene.
Claim 25
Dependent Claim
16 words
A DNA sequence according to claim 21 wherein the coding sequence codes for a signal peptide.
Claim 26
Dependent Claim
56 words
A chimeric DNA sequence comprising a first non-coding chemically regulatable DNA component sequence which is capable of regulating the transcription of an associated DNA sequence in a plant or plant tissue, wherein this regulation is dependent upon a chemical regulator, and a second DNA component sequence capable of being transcribed in a plant or plant tissue.
Claim 27
Dependent Claim
56 words
A chimeric DNA sequence comprising a first non-coding chemically regulatable DNA component sequence which is capable of regulating the transcription of an associated DNA sequence in a plant or plant tissue, wherein this regulation is dependent upon a chemical regulator, and a second DNA component sequence capable of being transcribed in a plant or plant tissue.
Claim 28
Dependent Claim
28 words
A chimeric DNA sequence according to claim 24 wherein said second DNA component sequence is capable of regulating the expression of a phenotypic trait by an anti-sense mechanism.
Claim 29
Dependent Claim
35 words
A chimeric DNA sequence according to claim 24 wherein said second DNA component sequence is a coding sequence that is capable of being transcribed and translated to produce a polypeptide resulting in a phenotypic trait.
Claim 30
Dependent Claim
21 words
A chimeric DNA sequence according to claim 24 wherein said second DNA component sequence is adjacent to the first component sequence.
Claim 31
Dependent Claim
21 words
A chimeric DNA sequence according to claim 24 wherein said first DNA component sequence is regulated by a repressing chemical regulator.
Claim 32
Dependent Claim
21 words
A chimeric DNA sequence according to claim 24 wherein said first DNA component sequence is regulated by an inducing chemical regulator.
Claim 33
Dependent Claim
36 words
A chimeric DNA sequence according to claim 24 wherein said first DNA component sequence is, or has substantial sequence homology to, a chemically regulatable DNA sequence in a naturally occurring chemically regulatable gene from a plant.
Claim 34
Dependent Claim
63 words
A chimeric DNA sequence according to claim 24 wherein said first DNA component sequence is, or has substantial sequence homology to, a chemically regulatable DNA sequence in a naturally occurring chemically regulatable gene from a plant and said second DNA component sequence is a coding sequence that is capable of being transcribed and translated to produce a polypeptide resulting in a phenotypic trait.
Claim 35
Dependent Claim
22 words
A chimeric DNA sequence according to claim 31 wherein said second DNA component sequence is adjacent to said first DNA coding sequence.
Claim 36
Dependent Claim
21 words
A chimeric DNA sequence according to claim 31 wherein said first DNA component sequence is regulated by an inducing chemical regulator.
Claim 37
Dependent Claim
31 words
A chimeric DNA sequence according to claim 31 wherein said first DNA component sequence is, or has substantial sequence homology to, the chemically inducible DNA sequence in a PR protein gene.
Claim 38
Dependent Claim
36 words
A chimeric DNA sequence according to claim 34 wherein said PR protein gene is a tobacco PR-1a, PR-1b, PR-1c, PR-1′, PR-Q or PR-R gene, a cucumber chitinase gene or a tobacco basic or acidic β-1,3-glucanase gene.
Claim 39
Dependent Claim
28 words
A chimeric DNA sequence according to claim 34 wherein said PR protein gene is a tobacco PR-1a, PR-1b, PR-1c, PR-1′ or PR-R gene or a cucumber chitinase gene.
Claim 40
Dependent Claim
20 words
A chimeric DNA sequence according to claim 34 wherein said PR protein gene is a tobacco PR-1a or PR-1′ gene.
Claim 41
Dependent Claim
19 words
A chimeric DNA sequence according to claim 34 wherein said PR protein gene is a basic tobacco β-1,3-glucanase gene.
Claim 42
Dependent Claim
38 words
A chimeric DNA sequence according to claim 31 wherein said first DNA component sequence is the 5′ flanking region of the tobacco PR-1a gene and contains about 300 or more base pairs adjacent to the transcriptional start site.
Claim 43
Dependent Claim
28 words
A chimeric DNA sequence according to claim 39 wherein said first DNA component sequence contains between about 600 and 1000 base pairs adjacent to the transcriptional start site.
Claim 44
Dependent Claim
19 words
A chimeric DNA sequence according to claim 31 wherein the second DNA component sequence codes for a phenotypic trait.
Claim 45
Dependent Claim
49 words
A chimeric DNA sequence according to claim 41 wherein said phenotypic trait is selected from the group consisting of tolerance or resistance to a herbicide, fungus, virus, bacterium, insect, nematode or arachnid; production of secondary metabolites; male or female sterility; and production of an enzyme or other reporter compound.
Claim 46
Dependent Claim
22 words
A chimeric DNA sequence according to claim 41 wherein said phenotypic trait is tolerance or resistance to herbicides or resistance to insects.
Claim 47
Dependent Claim
30 words
A chimeric DNA sequence according to claim 43 wherein said tolerance or resistance to herbicides is caused by a DNA sequence coding for wild-type or herbicide resistant acetohydroxyacid synthase (AHAS).
Claim 48
Dependent Claim
25 words
A chimeric DNA sequence according to claim 43 wherein said resistance to insects is caused by a DNA sequence coding for Bacillus thuringiensis endotoxin (BT).
Claim 49
Dependent Claim
46 words
A chimeric DNA sequence according to claim 41 wherein said phenotypic trait is a reporter compound selected from the group consisting of luciferase (LUX), chloramphenicol acetyltransferase (CAT), neomycin phosphotransferase (NPT), nopaline synthase (NOS), octopine synthase (OCS), beta-l,3-glucuronidase (GUS), acetohydroxyacid synthase (AHAS), and Bacillus thuringiensis endotoxin (BT).
Claim 50
Dependent Claim
32 words
A chimeric DNA sequence according to claim 41 wherein said phenotypic trait is a reporter compound selected from the group consisting of beta-l,3-glucuronidase (GUS), acetohydroxyacid synthase (AHAS), and Bacillus thuringiensis endotoxin (BT).
Claim 51
Dependent Claim
26 words
A chimeric DNA sequence according to claim 24 wherein said first DNA component sequence is regulated by a chemical regulator inducing PR protein genes in plants.
Claim 52
Dependent Claim
31 words
A chimeric DNA sequence according to claim 48 wherein the chemical regulator is selected from the group consisting of benzoic acid, salicylic acid, acetylsalicylic acid, polyacrylic acid and substituted derivatives thereof.
Claim 53
Dependent Claim
33 words
A chimeric DNA sequence according to claim 48 wherein the chemical regulator is selected from the group consisting of benzo-1,2,3-thiadiazolecarboxylic acid, benzo-1,2,3-thiadiazolethiocarboxylic acid, cyanobenzo-1,2,3-thiadiazole, benzo-1,2,3-thiadiazolecarboxylic acid amide, benzo-1,2,3-thiadiazolecarboxylic acid hydrazide, and derivatives thereof.
Claim 54
Dependent Claim
33 words
A chimeric DNA sequence according to claim 48 wherein the chemical regulator is selected from the group consisting of benzo-1,2,3-thiadiazole-7-carboxylic acid, benzo-1,2,3-thiadiazole-7-thiocarboxylic acid, 7-cyanobenzo-1,2,3-thiadiazole, benzo-1,2,3-thiadiazole-7-carboxylic acid amide, benzo-1,2,3-thiadiazole-7-carboxylic acid hydrazide, and derivatives thereof.
Claim 55
Dependent Claim
40 words
A chimeric DNA sequence according to claim 48 wherein the chemical regulator is selected from the group consisting of benzo-1,2,3-thiadiazolecarboxylic acid, alkyl benzo-1,2,3-thiadiazolecarboxylate in which the alkyl group contains one to six carbon atoms, and substituted derivatives of these compounds.
Claim 56
Dependent Claim
19 words
A chimeric DNA sequence according to claim 48 wherein the chemical regulator is dichloroisonicotinic acid or a derivative thereof.
Claim 57
Dependent Claim
35 words
A chimeric DNA sequence according to claim 48 wherein the chemical regulator is selected from the group consisting of benzo-1,2,3-thiadiazole-7-carboxylic acid, methyl benzo-1,2,3-thiadiazole-7-carboxylate, n-propyl benzo-1,2,3-thiadiazole-7-carboxylate, benzyl benzo-1,2,3-thiadiazole-7-carboxylate, benzo-1,2,3-thiadiazole-7-carboxylic acid sec-butylhydrazide, 2,6-dichloroisonicotinic acid, and methyl 2,6-dichloroisonicotinate.
Claim 58
Dependent Claim
15 words
A chimeric DNA sequence according to claim 48 wherein the chemical regulator is methyl benzo-1,2,3-thiadiazole-7-carboxylate.
Claim 59
Dependent Claim
118 words
A chimeric DNA sequence according to claim 24 comprising a first DNA component sequence which is, or has substantial sequence homology to, a non-coding, chemically regulatable DNA sequence of a naturally occurring chemically regulatable gene, this first DNA component sequence being capable of regulating the transcription of an associated DNA sequence in a plant or plant tissue, wherein this regulation is dependent upon a chemical regulator, a second DNA component sequence which is, or has substantial sequence homology to, part but not all of a transcribable DNA sequence with which the first component is associated in the naturally occurring chemically regulatable gene, and a third DNA component sequence capable of being transcribed in a plant or plant tissue.
Claim 60
Dependent Claim
31 words
A chimeric DNA sequence according to claim 56 wherein the first DNA component sequence is, or has substantial sequence homology to, the chemically inducible DNA sequence in a PR protein gene.
Claim 61
Dependent Claim
19 words
A chimeric DNA sequence according to claim 56 wherein the second DNA component sequence codes for a signal peptide.
Claim 62
Dependent Claim
34 words
A chimeric DNA sequence according to claim 58 wherein the second DNA component sequence codes for a peptide which is, or has substantial sequence homology to, the signal peptide from a PR protein gene.
Claim 63
Dependent Claim
17 words
A chimeric DNA sequence according to claim 59 wherein the PR protein gene is the PR-1a gene.
Claim 64
Dependent Claim
19 words
A chimeric DNA sequence according to claim 56 wherein the third DNA component sequence codes for a phenotypic trait.
Claim 65
Dependent Claim
10 words
A vector comprising the non-coding DNA sequence of claim 1.
Claim 66
Dependent Claim
14 words
A vector according to claim 62 comprising the chimeric DNA sequence of claim 24.
Claim 67
Dependent Claim
16 words
A vector according to claim 62 which is functional in plant cells or in Agrobacterium .
Claim 68
Dependent Claim
16 words
E . coli HB101/pTJS75-Km, ATCC number 67628, deposited on February 12, 1988, according to claim 62.
Claim 69
Dependent Claim
14 words
Plasmid pBS-PR1013Cla, ATCC number 40426, deposited on February 12, 1988, according to claim 62.
Claim 70
Dependent Claim
14 words
Plasmid pBS-pR1019, ATCC number 40427, deposited on February 12, 1988, according to claim 62.
Claim 71
Dependent Claim
14 words
Plasmid pBS-Gluc39.1, ATCC number 40526, deposited on December 29, 1988, according to claim 62.
Claim 72
Dependent Claim
14 words
Plasmid pBS-Gluc39.3, ATCC number 40527, deposited on December 29, 1988, according to claim 62.
Claim 73
Dependent Claim
14 words
Plasmid pBScucchi/chitinase, ATCC number 40528, deposited on December 29, 1988, according to claim 62.
Claim 74
Dependent Claim
15 words
Plasmid pBSGL6e, ATCC number 40535 , deposited on January 18, 1989, according to claim 62.
Claim 75
Dependent Claim
13 words
Plant tissue, plant or seed containing the chimeric DNA sequence of claim 24.
Claim 76
Dependent Claim
21 words
A substantially pure signal peptide having the amino acid sequence MetGlyPheValLeuPheSerGlnLeuProSerPheLeuLeuValSerThrLeuLeuPheLeuValIleSerHisSerCysArgAla or a peptide having substantial sequence homology to this peptide.
Claim 77
Dependent Claim
21 words
A substantially pure signal peptide having the amino acid sequence MetGlyPheValLeuPheSerGlnLeuProSerPheLeuLeuValSerThrLeuLeuPheLeuValIleSerHisSerCysArgAla or a peptide having substantial sequence homology to this peptide.
Claim 78
Dependent Claim
24 words
A DNA sequence which codes for the signal peptide of claim 73 or for a peptide having substantial sequence homology to that signal peptide.
Claim 79
Dependent Claim
26 words
A substantially pure DNA sequence according to claim 19 as shown in "Sequence 1" or having substantial sequence homology to the sequence shown in "Sequence 1".
Claim 80
Dependent Claim
26 words
A substantially pure DNA sequence according to claim 19 as shown in "Sequence 2" or having substantial sequence homology to the sequence shown in "Sequence 2".
Claim 81
Dependent Claim
22 words
A substantially pure DNA sequence as shown in "Sequence 3" or having substantial sequence homology to the sequence shown in "Sequence 3".
Claim 82
Dependent Claim
22 words
A substantially pure DNA sequence as shown in "Sequence 3" or having substantial sequence homology to the sequence shown in "Sequence 3".
Claim 83
Dependent Claim
22 words
A substantially pure DNA sequence as shown in "Sequence 4" or having substantial sequence homology to the sequence shown in "Sequence 4".
Claim 84
Dependent Claim
22 words
A substantially pure DNA sequence as shown in "Sequence 4" or having substantial sequence homology to the sequence shown in "Sequence 4".
Claim 85
Dependent Claim
26 words
A substantially pure DNA sequence according to claim 19 as shown in "Sequence 5" or having substantial sequence homology to the sequence shown in "Sequence 5".
Claim 86
Dependent Claim
26 words
A substantially pure DNA sequence according to claim 19 as shown in "Sequence 6" or having substantial sequence homology to the sequence shown in "Sequence 6".
Claim 87
Dependent Claim
22 words
A substantially pure DNA sequence as shown in "Sequence 7" or having substantial sequence homology to the sequence shown in "Sequence 7".
Claim 88
Dependent Claim
22 words
A substantially pure DNA sequence as shown in "Sequence 7" or having substantial sequence homology to the sequence shown in "Sequence 7".
Claim 89
Dependent Claim
22 words
A substantially pure DNA sequence as shown in "Sequence 8" or having substantial sequence homology to the sequence shown in "Sequence 8".
Claim 90
Dependent Claim
22 words
A substantially pure DNA sequence as shown in "Sequence 8" or having substantial sequence homology to the sequence shown in "Sequence 8".
Claim 91
Dependent Claim
105 words
A process for the preparation of a substantially pure chemically regulatable DNA sequence from a chemically regulatable gene in a naturally occurring system containing that gene, which process comprises the steps of (a) activating expression in said system of RNA from the chemically regulatable gene; (b) isolating said RNA; (c) differentially screening a genomic library for RNA generated from RNA isolated from said activated system as well as RNA generated from RNA isolated from a control system that is not activated; (d) isolating a genomic clone; (e) subcloning the chemically regulatable gene from said genomic clone; and (f) isolating the desired chemically regulatable DNA sequence.
Claim 92
Dependent Claim
105 words
A process for the preparation of a substantially pure chemically regulatable DNA sequence from a chemically regulatable gene in a naturally occurring system containing that gene, which process comprises the steps of (a) activating expression in said system of RNA from the chemically regulatable gene; (b) isolating said RNA; (c) differentially screening a genomic library for RNA generated from RNA isolated from said activated system as well as RNA generated from RNA isolated from a control system that is not activated; (d) isolating a genomic clone; (e) subcloning the chemically regulatable gene from said genomic clone; and (f) isolating the desired chemically regulatable DNA sequence.
Claim 93
Dependent Claim
12 words
A substantially pure DNA sequence isolated by the process of claim 83.
Claim 94
Dependent Claim
114 words
A process according to claim 83 wherein said system is a plant, which process comprises the steps of (a) activating expression in said plant of polyA+ RNA from the chemically regulatable gene; (b) isolating said polyA+ RNA; (c) constructing a cDNA library from said polyA+ RNA; (d) differentially screening said cDNA library with cDNA generated from RNA in a control plant that is not activated; (e) isolating cDNA clones that are chemically regulatable; (f) isolating a genomic clone from a genomic library of said plant using as a probe the cDNA clone of step (e); (g) subcloning the chemically regulatable gene from said genomic clone; and (h) isolating the desired chemically regulatable DNA sequence.
Claim 95
Dependent Claim
16 words
A process according to claim 85 wherein said chemically reguiatable gene is a PR protein gene.
Claim 96
Dependent Claim
23 words
A process according to claim 86 wherein said PR protein gene is activated in step (a) by a chemical inducer or a pathogen.
Claim 97
Dependent Claim
41 words
A process according to claim 87 wherein said chemical inducer is selected from the group consisting of benzoic acid, salicylic acid, acetylsalicylic acid, polyacrylic acid, benzo-1,2,3-thiadiazolecarboxylic acid, benzo-1,2,3-thiadiazolethiocarboxylic acid, cyanobenzo-1,2,3-thiadiazole, benzo-1,2,3-thiadiazolecarboxylic acid amide, benzo-1,2,3-thiadiazolecarboxylic acid hydrazide, dichloroisonicotinic acid, and derivatives thereof.
Claim 98
Dependent Claim
31 words
A process for regulating transcription of a chemically regulatable gene, which process comprises applying a chemical regulator to plant tissue, plant or seed containing the chimeric DNA sequence of claim 24.
Claim 99
Dependent Claim
40 words
A process according to claim 89 wherein said regulator is selected from the group consisting of benzoic acid, salicylic acid, acetylsalicylic acid, polyacrylic acid, benzo-1,2,3-thiadiazolecarboxylic acid, benzo-1,2,3-thiadiazolethiocarboxylic acid, cyanobenzo-1,2,3-thiadiazole, benzo-1,2,3-thiadiazolecarboxylic acid amide, benzo-1,2,3-thiadiazolecarboxylic acid hydrazide, dichloroisonicotinic acid, and derivatives thereof.
Claim 100
Dependent Claim
37 words
A process according to claim 89 wherein the first DNA component sequence of said chimeric DNA is a chemically inducible sequence which is, or has substantial sequence homology to, a chemically inducible sequence of a PR gene.
Claim 101
Dependent Claim
46 words
A process according to claim 89 wherein the chemically regulatable gene codes for a phenotypic trait selected from the group consisting of a developmental state, sterility, disease resistance, pest resistance, herbicide tolerance or resistance, production of a secondary metabolite and production of an assayable enzyme marker.
Claim 102
Dependent Claim
35 words
A process according to claim 92 wherein said enzyme marker is luciferase (LUX), chloramphenicol acetyltransferase (CAT), neomycin phosphotransferase (NPT), nopaline synthase (NOS), octopine synthase (OCS), beta-l,3-glucuronidase (GUS), acetohydroxyacid synthase (AHAS), or Bacillus thuringiensis endotoxin (BT).
Claim 103
Dependent Claim
21 words
A process according to claim 93 wherein said enzyme marker is beta-l,3-glucuronidase (GUS), acetohydroxyacid synthase (AHAS), or Bacillus thuringiensis endotoxin (BT).
Claim 104
Dependent Claim
45 words
A process for identifying a chemical regulator which comprises transforming a host with the chimeric DNA sequence of claim 31 or a vector containing said chimeric DNA sequence, applying a putative chemical regulator to the transformed host, and measuring the expression of the phenotypic trait.
Claim 105
Dependent Claim
16 words
A process according to claim 95 wherein the transformed host is plant tissue or plant cells.
Claim 106
Dependent Claim
38 words
A process according to claim 96 wherein the first DNA component sequence of said chimeric DNA is a chemically inducible sequence which is, or has substantial sequence homology to, a chemically inducible sequence of a PR protein gene.
Claim 107
Dependent Claim
21 words
A process according to claim 95 wherein the phenotypic trait coded for by the chimeric DNA is an assayable enzyme marker.
Claim 108
Dependent Claim
35 words
A process according to claim 98 wherein said enzyme marker is luciferase (LUX), chloramphenicol acetyltransferase (CAT), neomycin phosphotransferase (NPT), nopaline synthase (NOS), octopine synthase (OCS), beta-l,3-glucuronidase (GUS), acetohydroxyacid synthase (AHAS), or Bacillus thuringiensis endotoxin (BT).
Claim 109
Dependent Claim
83 words
A process for identifying a chemically regulatable DNA sequence which comprises the steps of (a) transforming a host with a putative chemically regulatable DNA sequence or a vector containing said sequence, a second DNA sequence which is a host-selectable gene marker and a third DNA sequence which codes for a phenotypic trait; (b) applying a chemical regulator to said transformed host; and (c) measuring the expression or selecting for change in expression of the phenotypic trait coded for by the third DNA sequence.
Claim 110
Dependent Claim
83 words
A process for identifying a chemically regulatable DNA sequence which comprises the steps of (a) transforming a host with a putative chemically regulatable DNA sequence or a vector containing said sequence, a second DNA sequence which is a host-selectable gene marker and a third DNA sequence which codes for a phenotypic trait; (b) applying a chemical regulator to said transformed host; and (c) measuring the expression or selecting for change in expression of the phenotypic trait coded for by the third DNA sequence.
Claim 111
Dependent Claim
19 words
A process according to claim 100 wherein said third DNA sequence is a host-selectable or an assayable gene marker.
Claim 112
Dependent Claim
16 words
A process according to claim 100 wherein the transformed host is plant tissue or plant cells.
Claim 113
Dependent Claim
23 words
A process according to claim 100 wherein the second or third DNA sequence is a gene marker for herbicide resistance or antibiotic resistance.
Claim 114
Dependent Claim
31 words
A process according to claim 100 wherein said gene marker is an antibiotic resistance gene selected from the group consisting of hygromycin, kanamycin, chloramphenicol, bleomycin, puromycin, lincomycin and methotrexate resistance genes.
Claim 115
Dependent Claim
18 words
A process according to claim 100 wherein the gene marker is an aminoglycoside phosphotransferase IV hygromycin resistance gene.
Claim 116
Dependent Claim
47 words
A process according to claim 100 wherein the third DNA sequence codes for an assayable enzyme marker selected from the group consisting of luciferase (LUX), chloramphenicol acetyltransferase (CAT), neomycin phosphotransferase (NPT), nopaline synthase (NOS), octopine synthase (OCS), beta-1,3-glucuronidase (GUS), acetohydroxyacid synthase (AHAS), and Bacillus thuringiensis endotoxin (BT).
Claim 117
Dependent Claim
33 words
A process according to claim 100 wherein the third DNA sequence codes for an assayable enzyme marker selected from the group consisting of beta-1,3-glucuronidase (GUS), acetohydroxyacid synthase (AHAS), and Bacillus thuringiensis endotoxin (BT).
Claim 118
Dependent Claim
14 words
A substantially pure chemically regulatable DNA sequence identified by the process of claim 100.
Claim 119
Independent Claim
75 words
A method for selecting transformed plant cells or tissue which have been subjected to transforming DNA comprising a first DNA component sequence that is chemically regulatable by a known chemical regulator and an associated second DNA sequence coding for a phenotypic marker, which process comprises the steps of (a) treating putative transformants with said known chemical regulator and (b) selecting transformants according to expression of the phenotypic trait coded for by the second DNA sequence.
Claim 120
Independent Claim
75 words
A method for selecting transformed plant cells or tissue which have been subjected to transforming DNA comprising a first DNA component sequence that is chemically regulatable by a known chemical regulator and an associated second DNA sequence coding for a phenotypic marker, which process comprises the steps of (a) treating putative transformants with said known chemical regulator and (b) selecting transformants according to expression of the phenotypic trait coded for by the second DNA sequence.
Claim 121
Dependent Claim
13 words
A method according to claim 109 wherein said phenotypic trait is antibiotic resistance.
Claim 122
Dependent Claim
22 words
A method according to claim 110 wherein said antibiotic resistance is resistance to hygromycin, kanomycin, chloroamphenicol, bleomycin, puromycin, lincoymcin, G418 and methotrexate.
Claim 123
Dependent Claim
15 words
A method according to claim 109 wherein said phenotypic trait is an assayable enzyme marker.
Claim 124
Dependent Claim
41 words
A method according to claim 112 wherein said enzyme marker is selected from the group consisting of luciferase (LUX), chloramphenicol acetyltransferase (CAT), neomycin phosphotransferase (NPT), nopaline synthase (NOS), octopine synthase (OCS), beta-1,3-glucuronidase (GUS), acetohydroxyacid synthase (AHAS), and Bacillus thuringiensis endotoxin (BT).
Claim 125
Dependent Claim
27 words
A method according to claim 113 wherein said enzyme marker is selected from the group consisting of beta-l,3-glucuronidase (GUS), acetohydroxyacid synthase (AHAS), and Bacillus thuringiensis endotoxin (BT).
Claim 126
Dependent Claim
44 words
An assay for beta-1,3-glucuronidase (GUS) activity which comprises reacting plant tissue extracts in samples of less than 0.5 ml containing 1.5 to 3 mM 4-methyl umbelliferyl glucuronide at around 37°C for 1 to 5 hours and determining the concentration of the fluorescent indicator released.
Claim 127
Dependent Claim
44 words
An assay for beta-1,3-glucuronidase (GUS) activity which comprises reacting plant tissue extracts in samples of less than 0.5 ml containing 1.5 to 3 mM 4-methyl umbelliferyl glucuronide at around 37°C for 1 to 5 hours and determining the concentration of the fluorescent indicator released.
Claim 128
Independent Claim
106 words
A method for the determination of the amount of a specific RNA in a solution of RNA comprising (a) labeling a RNA specific primer oligonucleotide of a length of between 12 and 18 nucleotides to high specific radioactivity, (b) hybridizing the RNA with the labeled oligonucleotide at a concentration of between 0.1 and 20 nM for between 2 minutes and 24 hours at a temperature around 37°C, (c) elongating the primer oligonucleotide with reverse transcriptase in the presence of nucleotide triphosphates at a concentration of between 0.003 and 1 mM for between 1 and 120 minutes, and (d) separating and visualizing the elongation products with autoradiography.
Claim 129
Independent Claim
106 words
A method for the determination of the amount of a specific RNA in a solution of RNA comprising (a) labeling a RNA specific primer oligonucleotide of a length of between 12 and 18 nucleotides to high specific radioactivity, (b) hybridizing the RNA with the labeled oligonucleotide at a concentration of between 0.1 and 20 nM for between 2 minutes and 24 hours at a temperature around 37°C, (c) elongating the primer oligonucleotide with reverse transcriptase in the presence of nucleotide triphosphates at a concentration of between 0.003 and 1 mM for between 1 and 120 minutes, and (d) separating and visualizing the elongation products with autoradiography.
Claim 130
Dependent Claim
73 words
A process for the preparation of a chemically regulatable chimeric DNA sequence comprising a first non-coding, chemically regulatable DNA component sequence which is capable of regulating the transcription of an associated DNA sequence in a plant or plant tissue, wherein this regulation is dependent upon a chemical regulator, and a second DNA component sequence capable of being transcribed in a plant or plant tissue, characterized in that the DNA component sequences are ligated.
Claim 131
Dependent Claim
73 words
A process for the preparation of a chemically regulatable chimeric DNA sequence comprising a first non-coding, chemically regulatable DNA component sequence which is capable of regulating the transcription of an associated DNA sequence in a plant or plant tissue, wherein this regulation is dependent upon a chemical regulator, and a second DNA component sequence capable of being transcribed in a plant or plant tissue, characterized in that the DNA component sequences are ligated.
Claim 132
Dependent Claim
132 words
A process for the prepartion of a chimeric DNA sequence comprising a first DNA component sequence which is, or has substantial sequence homology to, a non-coding, chemically regulatable DNA sequence of a naturally occurring chemically regulatable gene, this first DNA component sequence being capable of regulating the transcription of an associated DNA sequence in a plant or plant tissue, wherein this regulation is dependent upon a chemical regulator, a second DNA component sequence which is, or has substantial sequence homology to, part but not all of a transcribable DNA sequence with which the first component is associated in the naturally occurring chemically regulatable gene, and a third DNA component sequence capable of being transcribed in a plant or plant tissue, characterized in that the DNA component sequences are ligated concurrently or consecutively.
Claim 133
Dependent Claim
132 words
A process for the prepartion of a chimeric DNA sequence comprising a first DNA component sequence which is, or has substantial sequence homology to, a non-coding, chemically regulatable DNA sequence of a naturally occurring chemically regulatable gene, this first DNA component sequence being capable of regulating the transcription of an associated DNA sequence in a plant or plant tissue, wherein this regulation is dependent upon a chemical regulator, a second DNA component sequence which is, or has substantial sequence homology to, part but not all of a transcribable DNA sequence with which the first component is associated in the naturally occurring chemically regulatable gene, and a third DNA component sequence capable of being transcribed in a plant or plant tissue, characterized in that the DNA component sequences are ligated concurrently or consecutively.
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