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US 10,059,952 B2Granted
Yeast strains for protein production
AssigneeGlycoFi, Inc.
InventorJuergen Nett
Life of this patent
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Abstract
Method and system for expression systems, based on ade1 and ade2 auxotrophic strains of yeast and fungi, including P. pastoris are disclosed. The expression systems are useful for increased cellular productivity of transformed cell lines and for production of recombinant glycoproteins at industrial scale.
Claims9
A method for expressing a recombinant peptide, proteins, and/or nucleic acid of interest in a lower eukaryote host cell comprising:
- (a) providing the host cell in which the endogenous ADE2 gene has been removed from the genome of the host cell;
- and
- (b) transforming the host cell with an integration vector comprising:
- (1) a nucleic acid having an open reading frame (ORF) encoding Ade2p and which is operably linked to
- (i) a truncated endogenous ADE2 promoter that consists of 3′ 51 nucleotides or less,
- (ii) a truncated endogenous ADE2 promoter that consists of 3′ 13 nucleotides or less, or
- (iii) no promoter;
- (2) a nucleic acid having one or more expression cassettes comprising a nucleic acid encoding a peptide, polypeptide, protein, or functional nucleic acid of interest, wherein the functional nucleic acid is selected from the group consisting of ribozyme, antisense nucleic acid, and siRNA molecule;
- and
- (3) a targeting nucleic acid that directs insertion of the integration vector into a particular location of the genome of the host cell by homologous recombination, wherein the transformed host cell produces the recombinant peptide, protein and/or nucleic acid of interest.
7 more elements
Dependencies are inferred from the claim text.