Patent ID: 6068994
Filing Date: 2000-05-30
Classification: C07K,C12N

Abstract:
A method for expression in yeast of a heterologous protein to a level of at least 5% of total cell protein, wherein the protein comprises a destabilizing N-terminal amino acid residue selected from the group consisting of isoleucine, glutamic acid, histidine, tyrosine, glutamine, aspartic acid, asparagine, phenylalanine, leucine, tryptophan, lysine, and arginine, comprising:(a) providing a DNA molecule encoding a ubiquitin fusion protein, said DNA molecule comprising a first DNA sequence that encodes ubiquitin and is immediately adjacent to, upstream of and in reading frame with a second DNA sequence that encodes the heterologous protein,wherein said first and second DNA sequences are operably linked to a promoter capable of promoting expression of the fusion protein to a level of at least 5% of total cell protein;(b) transforming a yeast host cell with said DNA molecule; and(c) culturing the transformed yeast host cell under conditions capable of inducing the expression and quantitative in vivo processing of the fusion protein, thereby yielding expression of the heterologous protein to a level of at least 5% of total cell protein.