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US 9,133,464 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.
Claims12
An expression system comprising:
- (a) a Pichia host cell in which the ADE1 or ADE2 endogenous gene has been mutated;
- wherein the host cell is auxotrophic for adenine;
- and
- (b) an integration vector comprising:
- (1) a nucleic acid comprising an:
- (i) ADE2 gene operably linked to a truncated ADE2 promoter that consists of the 3′ 82 nucleotides of said promoter;
- (ii) ADE2 gene operably linked to a truncated ADE2 promoter that consists of the 3′ 13 nucleotides of said promoter;
- (iii) ADE1 gene not operably linked to a promoter;
- or
- (iv) ADE2 gene not operably linked to a promoter;
- (2) a nucleic acid having an insertion site for the insertion of one or more expression cassettes comprising a nucleic acid encoding a peptide, protein and/or functional nucleic acid interest, 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.
9 more elements
Dependencies are inferred from the claim text.