EP0159884A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Heat shock promoter and gene
Assignee: University of Georgia Research Foundation Inc
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Filing: 1985-04-12
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Granted: —
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Expires: 2005-04-12
Abstract
Abstract A bacterial strain, for example Escherichia coli, contains and replicates a recombinant DNA plasmid comprising a vector, a fragment of plant DNA controlling expression of a heat shock gene from an Agrobacterium strain inserted into a T-DNA fragment of a Ti-plasmid and a foreign gene or a soybean gene under control of said fragment of plant DNA. A suitable vector is pBR322, and examples of suitable foreign genes include a Z-gene coating for beta -galactosidase or a crystalline endotoxin gene. Also disclosed are methods for recognising plant cells transformed by T-DNA of Agrobacterium species by expressing a transformation recognition gene under control of a soybean heat shock gene promoter.
Inventors
- 👤 Joe L. Key
- 👤 William B. Gurley
- 👤 Ronald T. Nagao
- 👤 Friedrich Schoeffl
- 👤 Eva Czarnecka
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-08 05:08
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/82; C12N15/8216; C12N15/8237; C07; C07K; C07K14/00; C07K14/195; C07K14/32; C07K14/325
Claims Analysis
Total Claims: 20
Claim 1
Independent Claim
36 words
1 . A recombinant DNA plasmid comprising; (a) a vector, and (b) a fragment of plant DNA controlling expression of a heat shock gene inserted into a T-DNA fragment of a Ti-plasmid from an Agrobacterium strain.
Claim 2
Dependent Claim
36 words
1 . A recombinant DNA plasmid comprising; (a) a vector, and (b) a fragment of plant DNA controlling expression of a heat shock gene inserted into a T-DNA fragment of a Ti-plasmid from an Agrobacterium strain.
Claim 3
Dependent Claim
14 words
A recombinant DNA plasmid as recited in Claim 1 wherein said vector is pBR322.
Claim 4
Dependent Claim
24 words
A recombinant DNA plasmid as recited in Claim 1 or Claim 2 wherein said fragment of plant DNA is a fragment of soybean DNA.
Claim 5
Dependent Claim
17 words
Claims 1 to 3 4. A recombinant DNA plasmid as recited in any of/wherein said fragment of
Claim 6
Dependent Claim
30 words
Claims 1 to 3 5. A recombinant DNA plasmid as recited in any of / wherein said fragment of plant DNA is obtained from a soybean plant, the fragment thereof
Claim 7
Dependent Claim
29 words
Claims 1 to 3 6. A recombinant DNA plasmid as recited in any off wherein said fragment of plant DNA is obtained from a soybean plant, the fragment thereof
Claim 8
Dependent Claim
29 words
Claims 1 to 3 7 . A recombinant DNA plasmid as recited in any of/wherein said fragment of plant DNA is obtained from a soybean plant, the fragment thereof
Claim 9
Dependent Claim
24 words
A recombinant DNA plasmid as recited in any preceding claim further comprising a structural gene under the control of said fragment of plant DNA.
Claim 10
Dependent Claim
17 words
A recombinant DNA plasmid as recited in Claim 8 wherein said structural gene is a foreign gene.
Claim 11
Dependent Claim
19 words
A recombinant DNA plasmid as recited in Claim 9 wherein said foreign gene is a Z-gene coding for fl-galactosidase.
Claim 12
Dependent Claim
18 words
A recombinant DNA plasmid as recited in Claim 9 wherein said foreign gene is a crystalline endotoxin gene.
Claim 13
Dependent Claim
22 words
A recombinant DNA plasmid as recited in Claim 11 wherein said crystalline endotoxin gene is a crystalline endotoxin gene of Bacillus thuringiensis.
Claim 14
Dependent Claim
17 words
A recombinant DNA plasmid as recited in Claim 8 wherein said structural gene is a soybean gene.
Claim 15
Independent Claim
287 words
A method for expressing a structural gene under control of a soybean heat shock gene promoter fragment comprising the steps: (a) isolating said soybean heat shock gene promoter fragment, (b) cloning said soybean heat shock gene promoter fragment into a T-DNA shuttle vector producing a recombinant DNA plasmid, (c) isolating a DNA fragment carrying foreign structural genes or soybean genes and inserting said DNA fragment into said recombinant DNA plasmid at a position on the 3'-side of the reading strand of said soybean heat shock gene promoter producing a heat shock expression plasmid, wherein said DNA fragment is oriented with respect to said soybean heat shock gene promoter as to be expressible under control thereof, (d) transforming said heat shock expression plasmid into a first bacterial strain capable of supporting replication of said heat shock expression plasmid, (e) mixing said bacterial strain capable of supporting replication of said heat shock expression plasmid with a second bacterial strain carrying a helper plasmid capable of transporting said heat shock expression plasmid into a third bacterial strain incapable of supporting replication of said heat shock expression plasmid, said third bacterial strain carrying a resident plasmid, (f) selecting for recombination between said heat shock expression plasmid and said resident plasmid giving a recombinant resident plasmid (g) infecting a plant or plant cell cultures with said third bacterial strain containing and replicating therein said recombinant resident plasmid, and (h) selecting plants or plant cell cultures comprising plant cells containing said foreign structural genes or soybean genes under control of said soybean heat shock gene promoter transferred from said recombinant resident plasmid to said plant cells, said foreign structural genes or soybean genes being expressed following heat shock treatment or other stress treatment.
Claim 16
Independent Claim
287 words
A method for expressing a structural gene under control of a soybean heat shock gene promoter fragment comprising the steps: (a) isolating said soybean heat shock gene promoter fragment, (b) cloning said soybean heat shock gene promoter fragment into a T-DNA shuttle vector producing a recombinant DNA plasmid, (c) isolating a DNA fragment carrying foreign structural genes or soybean genes and inserting said DNA fragment into said recombinant DNA plasmid at a position on the 3'-side of the reading strand of said soybean heat shock gene promoter producing a heat shock expression plasmid, wherein said DNA fragment is oriented with respect to said soybean heat shock gene promoter as to be expressible under control thereof, (d) transforming said heat shock expression plasmid into a first bacterial strain capable of supporting replication of said heat shock expression plasmid, (e) mixing said bacterial strain capable of supporting replication of said heat shock expression plasmid with a second bacterial strain carrying a helper plasmid capable of transporting said heat shock expression plasmid into a third bacterial strain incapable of supporting replication of said heat shock expression plasmid, said third bacterial strain carrying a resident plasmid, (f) selecting for recombination between said heat shock expression plasmid and said resident plasmid giving a recombinant resident plasmid (g) infecting a plant or plant cell cultures with said third bacterial strain containing and replicating therein said recombinant resident plasmid, and (h) selecting plants or plant cell cultures comprising plant cells containing said foreign structural genes or soybean genes under control of said soybean heat shock gene promoter transferred from said recombinant resident plasmid to said plant cells, said foreign structural genes or soybean genes being expressed following heat shock treatment or other stress treatment.
Claim 17
Dependent Claim
13 words
A method as recited in Claim 14 wherein said heat shock gene promoter
Claim 18
Dependent Claim
13 words
A method as recited in Claim 14 wherein said heat shock gene promoter
Claim 19
Independent Claim
24 words
A method as recited in any of Claims 14 to 16 wherein said foreign structural gene is a crystalline toxic protein of Bacillus thuringiensis.
Claim 20
Independent Claim
21 words
A method as recited in any of Claims 14 to 16 wherein said foreign structural gene is a herbicide resistance gene.
References Cited (Backward Citations)
No citation records indexed.
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