Compositions and methods for improved protein production
AssigneeDanisco US Inc.
InventorsElizabeth A. Bodie, Steve Kim
Life of this patent
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Abstract
The present invention relates to the identification of novel nucleic acid sequences, designated herein as 7p, 8k, 7E, 9G, 8Q and 203, in a host cell which effect protein production. The present invention also provides host cells having a mutation or deletion of part or all of the gene encoding 7p, 8k, 7E, 9G, 8Q and 203, which are presented in FIG. 1, and are SEQ ID NOS.: 1-6, respectively. The present invention also provides host cells further comprising a nucleic acid encoding a desired heterologous protein such as an enzyme.
Claims19
A filamentous fungus having a mutation or deletion of part or all of a gene having the sequence selected from at least one sequence set forth in any one of SEQ ID NOs:1-6, and said mutation or deletion results in the enhanced production of a desired polypeptide compared to the parent filamentous fungus.
An isolated nucleotide sequence selected from a group consisting of SEQ ID NOs:
- 1-6.
(canceled)
A method for producing a novel strain of T. reesei using insertional mutagenesis wherein said novel strain of T. reesei has enhanced cellulase production as compared to the parent strain of T. reesei, comprising:
- (a) preparing a population of competent Agrobacterium sp. cells by electroporating into competent Agrobacterium sp. cells an expression vector comprising, in operable condition, the left and right T-DNA boarder regions, pV51 plasmid origins for replication in Agrobacterium sp. and bacterial markers to confer resistance to chloramphenicol to create a population comprising transformed Agrobacterium sp. cells;
- (b) selecting for Agrobacterium sp. from said population of step (a);
- (c) inoculating a culture of T. reesei spores with the Agrobacterium sp. transformants of step (b) to create an induction culture;
- (d) culturing said induction culture of step (c) at about 18° C. and for about 24 hours to create a population;
- (e) transferring samples of said population of transformed T. reesei of step (d) to selective medium and isolating colonies of T. reesei effective in degrading cellulose;
- and
- (f) comparing the effectiveness of cellulose degradation between the T. reesei of the isolated colonies of step (e) and the non-transformed parent strain, wherein said T. reesei of the isolated colonies of step (e) are enhanced to in cellulose degradation when compared to the non-transformed parent strain.
Dependencies are inferred from the claim text.