EXPRESSION AND HIGH-THROUGHPUT SCREENING OF COMPLEX EXPRESSED DNA LIBRARIES IN FILAMENTOUS FUNGI

摘要

The invention is generally directed to modified filamentous fungal host cells comprising one or more nucleic acids encoding one or more polypeptides under the control of one or more promoters that are functional in said cells. Methods of using the modified cells to express one or more polypeptides are also disclosed, including methods of screening cells transformed with one or more expression vectors comprising nucleic acids derived from synthetic or genomic nucleic acids including, cDNAs. Methods of purifying one or more polypeptides or complexes comprising one or more polypeptides expressed in the modified cells, intended for use as substrates in structure/function studies, as therapeutic agents, as diagnostic reagents, or as human or animal vaccines, are also disclosed.

主权项

1. A modified cell from a filamentous fungus comprising one or more nucleic acids, each nucleic acid comprising at least one coding region encoding at least one polypeptide, wherein at least one coding region is operably-linked to at least one regulatory region comprising at least one promoter active in said cell;
wherein said fungus is in a genus selected from the group consisting of Aspergillus, Fusarium, Chrysosporium, Myceliophthora, and Trichoderma; wherein said fungus has been modified to have less than 50% of the protease activity of a parental or non-modified fungus; wherein said fungus has a phenotype characterized by a culture viscosity of less than 200 cP at the end of fermentation, when cultured in suspension, in the presence of adequate nutrients, under optimal or near-optimal conditions.