Claims
- 1. A transcription factor regulating an α-amylase promoter initiated expression in filamentous fungus.
- 2. The factor of claim 1 originating from a fungus of the genus Aspergillus, Trichoderma, Penicillium, Fusarium, Humicola, etc.
- 3. The factor of claim 2 originating from the species A. oryzae, A. niger, A. awamori, especially A. oryzae IFO4177.
- 4. The factor of claim 3 comprising one or more truncated forms of the factor wherein the factor comprises the amino acid sequence depicted as SEQ ID NO: 3.
- 5. A DNA construct having a DNA sequence coding for the factor of any of the claims 1 to 4.
- 6. The DNA sequence of claim 5 comprising one or more fragments or a combination of fragments of the DNA sequence depicted as SEQ ID NO: 1 and SEQ ID NO: 2.
- 7. A DNA construct comprising a DNA sequence encoding a transcription factor exhibiting activity in regulating the expression of an α-amylase promoter in a filamentous fungus, which DNA sequence comprises
a) the transcription factor encoding part of the DNA sequence cloned into plasmid pToC320 present in E. coli ToC1058, DSM 10666, or b) an analogue of the DNA sequence defined in a), which
i) is at least 60% homologous with the DNA sequence defined in a), or ii) hybridizes with the same nucleotide probe as the DNA sequence defined in a), or iii) encodes a transcription factor which is at least 50% homologous with the transcription factor encoded by a DNA sequence comprising the DNA sequence defined in a), or iv) encodes a transcription factor which is immunologically reactive with an antibody raised against the purified transcription factor encoded by the DNA sequence defined in a), or v) complements the mutation in DSM No. 10671, i.e. makes DSM No. 10671 able to grow on cyclodextrin and produce lipase when transformed with said DNA sequence.
- 8. The DNA construct of any of the claims 5 to 7, in which said DNA sequence is obtainable from a filamentous fungus.
- 9. The DNA construct of claim 8, in which said filamentous fungus belongs to any of the groups Phycomycetes, Zygomycetes, Ascomycetes, Basidiomycetes and fungi imperfecti, including Hyphomycetes such as the genera Aspergillus, Penicillium, Trichoderma, Fusarium and Humicola, in particular a strain from Aspergillus sp., and especially from A. oryzae.
- 10. The DNA construct of claim 9, in which said DNA sequence is isolated from or produced on the basis of a DNA library of an Aspergillus oryzae strain.
- 11. The DNA construct of claims 5 to 8, in which said DNA sequence is obtainable from a yeast strain, especially of, Saccharomyces.
- 12. The DNA construct of claim 7, in which the DNA sequence is isolated from Eschericia coli DSM 10666.
- 13. A recombinant expression vector comprising a DNA construct of any of claims 5 to 12.
- 14. A cell comprising a DNA construct of any of claims 5 to 12, or a recombinant expression vector of claim 13.
- 15. The cell of claim 14, which is a eukaryotic cell, in particular a fungal cell, such as a yeast cell or a filamentous fungal cell.
- 16. The cell of claim 15, which is a strain of Aspergillus sp, in particular a strain of A. niger or A. oryzae.
- 17. The cell of claim 15, which is a strain of Trichoderma sp., in particular T. reesei.
- 18. The cell of claim 15, which is a strain of Saccharomyces, in particular a strain of S. cerevisiae.
- 19. A method of producing a polypeptide of interest comprising growing a cell of any of the claims 14 to 18 under conditions conducive to the production of said factor and said polypeptide of interest, and recovering said polypeptide of interest.
- 20. The method of claim 19, wherein said fungus is a fungus of the genus Aspergillus, Trichoderma, Penicillium, Fusarium or Humicola.
- 21. The method of claim 20, wherein said cell is of the species A. oryzae, A. niger, or A. awamori.
- 22. The method of claim 19, 20, or 21, wherein said polypeptide of interest is a medicinal polypeptide.
- 23. The method of claim 22, wherein said medicinal polypeptide is a growth hormone, insulin, or a blood clotting factor.
- 24. The method of claim 19, 20, or 21, wherein said polypeptide is an industrial enzyme.
- 25. The method of claim 24, wherein said industrial enzyme is a carbonyl hydrolase, carbohydrase, protease, lipase, amylase, cellulase, oxido reductase, glucoamylase, or esterase.
- 26. Use of a factor of any of the claims 1 to 4 for enhancing the expression of a polypeptide of interest in a filamentous fungus.
- 27. The use of claim 26, wherein said factor is the factor of claim 4.
- 28. The use of claim 27, wherein said fungus is a fungus of the genus Aspergillus, Penicillium, Trichoderma, Fusarium and Humicola, in particular a strain from Aspergillus sp., and especially from A. oryzae sp.
- 29. The use of claim 28, wherein said fungus is of the species A. oryzae, A. niger, A. awamori, T. reesei, or T. harzianum.
- 30. The use of any of the claims 26 to 29, wherein said polypeptide of interest is a medicinal polypeptide.
- 31. The use of claim 30, wherein said medicinal polypeptide is a growth hormone, insulin, or blood clotting factor.
- 32. The use of any of the claims 26 to 29, wherein said polypeptide is an industrial enzyme.
- 33. The use of claim 32, wherein said industrial enzyme is a carbonyl hydrolase, carbohydrase, protease, lipase, amylase, cellulase, oxido reductase, glucoamylase, or esterase.
- 34. A transcription factor regulating an -amylase promoter initiated expression in filamentous fungus.
- 35. The factor of claim 34 originating from Aspergillus, Fusarium, Humicola, Penicillium, or Trichoderma.
- 36. The factor of claim 35 originating from Fusarium.
- 37. The factor of claim 35 originating from A. awamori, A. niger, or A. oryzae.
- 38. The factor of claim 37 originating from A. oryzae IFO4177.
- 39. The factor of claim 38 comprising one or more truncated forms of the factor, wherein the factor comprises the amino acid sequence of SEQ. ID. NO:3.
- 40. A DNA construct having a DNA sequence coding for the factor of claim 34.
- 41. The DNA construct of claim 40 comprising one or more fragments, or a combination of fragments of the DNA sequences of SEQ ID NO:1 and SEQ ID NO:2.
- 42. The DNA construct of claim 40, wherein the DNA sequence originates from a filamentous fungus.
- 43. The DNA construct of claim 40, wherein the filamentous fungus is an Ascomycetes, Basidiomycetes Phycomycetes, Zygomycetes, or fungi imperfecti.
- 44. A recombinant expression vector comprising the DNA construct of claim 40.
- 45. A cell comprising the expression vector of claim 44.
- 46. The cell of claim 45, wherein the cell is a strain of Aspergillus.
- 47. The cell of claim 45, wherein the cell is a strain of Trichoderma.
- 48. The cell of claim 45, wherein the cell is a strain of Fusarium.
- 49. A method of producing a polypeptide of interest comprising growing the cell of claim 47 under conditions conducive to the production of the factor and the polypeptide of interest, and recovering the polypeptide of interest.
- 50. The method of claim 49, wherein the cell is a Aspergillus, Fusarium, Humicola, Penicillium, or Trichoderma cell.
- 51. The method of claim 50, wherein the cell is a Fusarium cell.
Priority Claims (1)
Number |
Date |
Country |
Kind |
0740/96 |
Jul 1996 |
DK |
|
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional of U.S. application Ser. No. 09/197,814 filed Nov. 23, 1998 which is a continuation of PCT/DK97/00305 filed Jul. 7, 1997 (the international application was published under PCT Article 21(2) in English) and claims, under 35 U.S.C. 119, priority of Danish application no. 0740/96 filed Jul. 5, 1996, the contents of which are fully incorporated herein by reference.
Divisions (1)
|
Number |
Date |
Country |
Parent |
09197814 |
Nov 1998 |
US |
Child |
09920581 |
Aug 2001 |
US |
Continuations (1)
|
Number |
Date |
Country |
Parent |
PCT/DK97/00305 |
Jul 1997 |
US |
Child |
09197814 |
Nov 1998 |
US |