Proteins from yeast host strains with defects in N-glycosylation
AssigneeBoehringer Mannheim GmbH
InventorsLudwig Lehle, Klaus Lehnert, Erhard Kopetzki
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Abstract
Saccharomyces mutants with defects in N-glycosylation which are obtainable by .sup.3 H!-mannose suicide selection, introduction of one or several selective markers, selection of those strains which, after transformation with the plasmid YEpL/glucose oxidase, secrete 10 mg/l GOD or more into the culture medium after culture under standard conditions, are allelic to the ngd mutations in Saccharomyces cerevisiae, DSM 7042, DSM 7338, DSM 7160 and/or 7340 and express proteins with a uniform carbohydrate structure.
Claims5
A hypoglycosylated recombinant protein obtained by transformation of a Saccharomyces cerevisiae mutant with DNA which codes for the desired protein, said mutant being of a phenotype which is not temperature sensitive, said mutant having defects in N-glycosylation resulting from alteration in the ngd 29 and/or ngd 62 genes and yielding a diploid having defects in the N-glycosylation when crossed with a strain of Saccharomyces cerevisiae selected from the group consisting of DSM 7042, DSM 7338, DSM 7160, and DSM 7340, fermentation of the cells of said transformed mutant, and isolation of said protein from the cells or from the culture supernatant, wherein said protein would be hyperglycosylated if expressed in wild-type Saccharomyces cerevisiae.
Dependencies are inferred from the claim text.