Filamentous fungi with inactivated protease genes for altered protein production
AssigneeDanisco US Inc., Genencor Division
InventorHuaming Wang
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Abstract
The invention relates to a filamentous fungal cell (e.g., Aspergillus sp.) comprising at least one inactivated protease gene chosen from apsB, a homolog of apsB, cpsA, a homolog cpsA, and combinations thereof. Nucleic acids and methods for making the inactivated mutant filamentous fungal cells are provided as well as methods for using the cells for the altered production of endogenous or heterologous proteins of interest.
Claims65
A filamentous fungal cell comprising at least one inactivated gene, wherein the inactivated gene is chosen from apsB, a homolog of apsB, cpsA, a homolog of cpsA, and combinations thereof.
A filamentous fungal cell comprising at least one inactivated gene, wherein the inactivated gene encodes an intracellular protein, and wherein production of at least one other protein by the cell is at least about 10% to about 60% greater than the production of the other protein in a corresponding parent strain of the filamentous fungal cell.
A method for producing a protein comprising:
- a) introducing a nucleic acid encoding a protein into a filamentous fungal cell, wherein said cell comprises at least one inactivated gene chosen from:
- apsB, a homolog of apsB, cpsA, a homolog cpsA, and combinations thereof;
- and
- b) growing the cell under conditions suitable for producing the protein.
A method for making a filamentous fungal strain for protein production comprising:
- a) transforming a filamentous fungal cell with a disruption sequence, wherein the disruption sequence comprises at least one inactivated gene chosen from:
- apsB, a homolog of apsB, cpsA, a homolog cpsA, and combinations thereof;
- and
- b) selecting the transformed cells wherein said disruption sequence is chromosomally integrated.
An isolated nucleic acid comprising a disruption sequence of a gene, wherein the gene is cpsA or a homolog of cpsA and wherein the disruption sequence comprises a selectable marker gene sequence reversely inserted at a restriction site in the coding region sequence of the gene.
An isolated nucleic acid comprising a disruption sequence of a gene, wherein the gene is apsB or a homolog of cpsA and wherein the disruption sequence comprises a selectable marker gene sequence reversely inserted at a restriction site in the coding region sequence of the gene.
Dependencies are inferred from the claim text.