CA2379498A1 - Plant gene expression, controlled by constitutive plant v-atpase promoters - Google Patents

Plant gene expression, controlled by constitutive plant v-atpase promoters Download PDF

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Publication number
CA2379498A1
CA2379498A1 CA002379498A CA2379498A CA2379498A1 CA 2379498 A1 CA2379498 A1 CA 2379498A1 CA 002379498 A CA002379498 A CA 002379498A CA 2379498 A CA2379498 A CA 2379498A CA 2379498 A1 CA2379498 A1 CA 2379498A1
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Prior art keywords
plant
dna construct
promoter
protoplast
atpase
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Granted
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CA002379498A
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French (fr)
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CA2379498C (en
Inventor
Elke Duwenig
Thomas Rausch
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BASF Plant Science GmbH
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Basf Plant Science Gmbh
Elke Duwenig
Thomas Rausch
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Publication of CA2379498A1 publication Critical patent/CA2379498A1/en
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
    • C12N15/8237—Externally regulated expression systems
    • C12N15/8238—Externally regulated expression systems chemically inducible, e.g. tetracycline
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
    • C12N15/8237—Externally regulated expression systems
    • C12N15/8239—Externally regulated expression systems pathogen inducible
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)

Abstract

To express selection and resistance genes, it is desirable to have plant constitutive promoters available which have strong, homogenous constitutive activity, if possible in all plant tissues or cell types and which, in addition, become more active or cannot be repressed in conditions of stress.
The invention concerns DNA syntheses which contain a plant V-ATPase promoter which is operatively bound to a heterologous gene. The invention also relates to the use of these syntheses in the form of expression cassettes, recombinant vectors and in transgenic plants, plant cells or protoplasts. The invention relates, in particular, to the promoter of the Beta vulgaris V-ATPase subunit c isoform 2.

Claims (52)

1 Claims 1. A DNA construct with the plant B. vulgaris V-ATPase promoter subunit c isoform 2 [SEQ ID No.1] or its functional equivalent, operatively linked with a heterologous gene.
2. A DNA construct as claimed in claim 1, wherein the nucleotide sequence of the plant B. vulgaris V-ATPase promoter subunit c isoform 2 [SEQ ID No.1] contains a deletion or wherein the promoter sequence is a hybrid of the B. vulgaris V-ATPase promoter subunit c isoform 2 sequence [SEQ ID No.1] and of the sequences of the other homologous V-ATPase promoters.
3. A DNA construct as claimed in claim 1 or 2, wherein the plant B. vulgaris V-ATPase promoter subunit c isoform 2 [SEQ ID No.1] is derived from dicotyledonous plants.
4. A DNA construct as claimed in claim 1 or 2, wherein the plant B. vulgaris V-ATPase promoter subunit c isoform 2 [SEQ ID No.1] is derived from monocotyledonous plants.
5. A DNA construct as claimed in claim 3 or 4, wherein the plant V-ATPase promoter is derived from sugar beet, tobacco, barley, rice, potatoes, sunflowers, soya, tomatoes, Canola, wheat, oilseed rape, sorghum, carrots, maize, Mesembranthemum crystallinum or Arabidopsis thaliana.
6. A DNA construct as claimed in any of claims 1 to 5, which encompasses a second promoter which can be regulated in a different manner than the first promoter.
7. A DNA construct as claimed in any of claims 1 to 6, wherein at least one further pyrimidine stretch is inserted in the plant promoter.
8. A DNA construct as claimed in any of claims 1 to 7, which is an expression cassette.
9. A DNA construct as claimed in any of claims 1 to 8, wherein the heterologous gene is a selection marker or a resistance-mediating gene or a gene of other medicinal, agronomical or other interest.
10. A polynucleotide encompassing the sequence of the promoter of B. vulgaris V-ATPase subunit c isoform 2 [SEQ ID No.1] or its functional equivalent.
11. The polynucleotide as claimed in claim 11, encompassing the sequence of a deleted or hybrid B. vulgaris V-ATPase subunit c isoform 2 promoter [SEQ ID No.1] or its functional equivalent.
12. A recombinant vector which encompasses a DNA construct as claimed in any of claims 1 to 9.
13. A recombinant vector as claimed in claim 12, which is a shuttle vector.
14. A recombinant vector as claimed in claim 12 or 13, which is an expression vector.
15. A microorganism which is transformed with a recombinant vector as claimed in any of claims 12 to 14.
16. A transgenic plant cell or protoplast whose genome encompasses a DNA
construct as claimed in any of claims 1 to 9.
17. A transgenic plant cell or protoplast as claimed in claim 16 derived from a monocotyledonous plant.
18. A transgenic plant cell or protoplast as claimed in claim 16 derived from a dicotyledonous plant.
19. A transgenic plant whose genome encompasses a DNA construct as claimed in any of claims 1 to 10.
20. A transgenic plant as claimed in claim 19, which is a monocotyledonous plant.
21. A transgenic plant as claimed in claim 19, which is a dicotyledonous plant.
22. A transgenic plant as claimed in claim 20 or 21, which is sugar beet, tobacco, barley, rice, potato, sunflower, soya, tomato, Canola, wheat, oilseed rape, sorghum, carrot, maize, Mesembranthemum crystallinum or Arabidopsis thaliana.
23. A method for the controlled expression of a heterologous gene in a plant cell or a protoplast, which comprises transforming the cell or the protoplast with a DNA construct as claimed in any of claims 1 to 9 and subsequently exposing the transformed cell or the protoplast to such a biotic or abiotic stress that the expression of the heterologous gene which has been transformed by means of the DNA construct is controlled.
24. The method as claimed in claim 23, wherein the plant cell or the protoplast is derived from a monocotyledonous plant.
25. The method as claimed in claim 23, wherein the plant cell or the protoplast is derived from a dicotyledonous plant.
26. The process as claimed in claim 24 or 25, wherein the plant cell or the protoplast is derived from sugar beet, tobacco, barley, rice, potatoes, sunflowers, soya, tomatoes, Canola, wheat, oilseed rape, sorghum, carrots, maize, Mesembranthemum crystallinum or Arabidopsis thaliana.
27. A method for the controlled expression of a heterologous gene in a plant, which comprises regenerating cells or protoplasts transformed with a DNA construct as claimed in any of claims 1 to 9 to give rise to a transgenic plant and subsequently exposing the plant transformed in this way to such a biotic or abiotic stress that the expression of the heterologous gene which has been transformed by means of the DNA construct is controlled.
28. The method as claimed in claim 27, wherein the transgenic plant is a monocotyledonous plant.
29. The method as claimed in claim 27, wherein the transgenic plant is a dicotyledonous plant.
30. Method as claimed in claim 28 or 29, wherein the transgenic plant is sugar beet, tobacco, barley, rice, potato, sunflower, soya, tomato, Canola, wheat, oilseed rape, sorghum, carrot, maize, Mesembranthemum crystadlinum or Arabidopsis thaliana.
31. A method for producing a recombinant protein, which comprises transforming a plant cell or a protoplast with a DNA construct as claimed in any of claims 1 to 9 and subsequently exposing the transformed cell or protoplast to such a biotic or abiotic stress that the recombinant protein transformed by means of the DNA construct is expressed.
32. The method as claimed in claim 31, wherein the plant cell or the protoplast is derived from a monocotyledonous plant.
33. The method as claimed in claim 31, wherein the plant cell or the protoplast is derived from a dicotyledonous plant.
34. The process as claimed in claim 32 or 33, wherein the plant cell or the protoplast is derived from sugar beet, tobacco, barley, rice, potatoes, sunflowers, soya, tomatoes, Canola, wheat, oilseed rape, sorghum, carrots, maize, Mesembranthemum crystallinum or Arabidopsis thaliana.
35. A method of producing a recombinant protein in a plant, which comprises regenerating cells or protoplasts transformed with a DNA construct as claimed in any of claims 1 to 9 to give rise to a transgenic plant and subsequently exposing the resulting transgenic plant to such. a biotic or abiotic stress that the recombinant protein transformed by means of the DNA construct is expressed.
36. The method as claimed in claim 35, wherein the transgenic plant is a monocotyledonous plant.
37. The method as claimed in claim 35, wherein the transgenic plant is a dicotyledonous plant.
38. Method as claimed in claim 36 or 37, wherein the transgenic plant is sugar beet, tobacco, barley, rice, potato, sunflower, soya, tomato, Canola, wheat, oilseed rape, sorghum, carrot, maize, Mesembranthemum crystallinum or Arabidopsis thaliana.
39. The use of a DNA construct as claimed in any of claims 1 to 9 for producing a recombinant protein in a plant cell or a protoplast.
40. The use of a DNA construct as claimed in any of claims 1 to 9 for producing a recombinant protein in a plant.
41. A recombinant protein prepared as claimed in any of claims 31 to 38.
42. The use of a DNA construct as claimed in any of claims 1 to 9 for expressing a gene in a plant under biotic or abiotic stress.
43. The use of a plant V-ATPase promoter for expressing a gene in a plant under biotic or abiotic stress.
44. The use as claimed in claim 43, wherein the plant B. vulgaris V-ATPase promoter subunit c isoform 2 contains a deletion or wherein the promoter sequence is a hybrid of the B. vulgaris V-ATPase promoter subunit c isoform 2 sequence [SEQ ID
No.1]
and of the sequences of the other homologous V-ATPase promoters.
45. The use as claimed in claim 44, wherein the plant V-ATPase promoter is derived from dicotyledonous plants.
46. The use as claimed in claim 44, wherein the plant V-ATPase promoter is derived from monocotyledonous plants.
47. The use as claimed in claim 45 or 46, wherein the plant V-ATPase promoter is derived from sugar beet, tobacco, barley, rice, potatoes, sunflowers, soya, tomatoes, Canola, wheat, oilseed rape, sorghum, carrots, maize, Mesembranthemum crystallinum or Arabidopsis thaliana.
48. The use as claimed in one of claims 43 to 47, wherein at least one further pyrimidine stretch is inserted into the plant promoter.
49. A plant cell or protoplast which is transformed with a DNA construct as claimed in any of claims 1 to 9 and which is resistant to biotic or abiotic stress.
50. A plant cell or protoplast as claimed in claim 49 which is resistant to salt stress.
51. A plant which is transformed with a DNA construct as claimed in any of claims 1 to 9 and which is resistant to biotic or abiotic stress.
52. A plant as claimed in claim 51 which is resistant to salt stress.
CA2379498A 1999-08-26 2000-08-10 Plant gene expression, controlled by constitutive plant v-atpase promoters Expired - Fee Related CA2379498C (en)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
US15088799P 1999-08-26 1999-08-26
DE19940648.0 1999-08-26
DE19940648 1999-08-26
US60/150,887 1999-08-26
PCT/EP2000/007774 WO2001014572A2 (en) 1999-08-26 2000-08-10 Plant gene expression, controlled by constitutive plant v-atpase promoters

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CA2379498A1 true CA2379498A1 (en) 2001-03-01
CA2379498C CA2379498C (en) 2012-12-18

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EP (1) EP1206560B1 (en)
JP (1) JP2003507074A (en)
CN (1) CN1371425A (en)
AR (1) AR033649A1 (en)
AT (1) ATE309381T1 (en)
AU (1) AU780117B2 (en)
BR (1) BR0013537A (en)
CA (1) CA2379498C (en)
DE (1) DE50011579D1 (en)
HU (1) HUP0203693A2 (en)
IL (1) IL147950A0 (en)
MX (1) MXPA02001786A (en)
PL (1) PL354490A1 (en)
RU (1) RU2002107795A (en)
WO (1) WO2001014572A2 (en)

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US8247652B2 (en) 2002-03-19 2012-08-21 Metanomics Gmbh & Co. Kgaa Population of transgenic plants individually comprising distinct codogenic gene segments, the population having at least 50% of the codogenic gene segments from a donor organism
US8952217B2 (en) 2005-10-14 2015-02-10 Metanomics Gmbh Process for decreasing verbascose in a plant by expression of a chloroplast-targeted fimD protein

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EP1206560B1 (en) 2005-11-09
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DE50011579D1 (en) 2005-12-15
HUP0203693A2 (en) 2003-04-28
CA2379498C (en) 2012-12-18
AU7509100A (en) 2001-03-19
PL354490A1 (en) 2004-01-26
CN1371425A (en) 2002-09-25
AR033649A1 (en) 2004-01-07
AU780117B2 (en) 2005-03-03
WO2001014572A2 (en) 2001-03-01
MXPA02001786A (en) 2003-07-14
EP1206560A2 (en) 2002-05-22
WO2001014572A3 (en) 2001-10-11
ATE309381T1 (en) 2005-11-15
RU2002107795A (en) 2004-03-10
BR0013537A (en) 2002-04-30

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