Synthesis, secretion and processing of alpha-factor-interferon fusion proteins in yeast.
A gene fusion consisting of 960 base pairs of 5'-flanking region of the yeast MF alpha 1 gene, 257 base pairs coding for alpha-factor prepro sequence, and a modified human IFN-alpha 1 gene was constructed. MAT alpha cells containing the chimeric gene synthesized and secreted active IFN-alpha 1 into the growth medium. The secreted interferon molecules contained the last 4 amino acids of alpha-factor prepro sequence and the amino acids encoded by the DNA modifications introduced at the beginning of IFN-alpha 1 gene. DNA sequences coding for these amino acids were removed by oligonucleotide-directed in vitro mutagenesis. Yeast cells transformed with expression plasmids containing the altered junction synthesized and secreted human IFN-alpha 1 with the natural NH2-terminus.
D001483:Base Sequence; D002678:Chimera; D002872:Chromosome Deletion; D003090:Coliphages / Q000235:genetics; D004262:DNA Restriction Enzymes; D004926:Escherichia coli / Q000235:genetics; D005796:Genes; D005800:Genes, Fungal; D006801:Humans; D007372:Interferons / Q000235:genetics*; D000072235:Mating Factor; D009154:Mutation; D009690:Nucleic Acid Conformation; D010455:Peptides / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014169:Transformation, Bacterial
D010455:Peptides; D000072235:Mating Factor; D007372:Interferons; D004262:DNA Restriction Enzymes