Open AccessSynthesis, secretion and processing of α-factor-interferon fusion proteins Id yeast
Author(s) -
Arjun Singh,
June M. Lugovoy,
William J. Kohr,
Perry Lj
Publication year - 1984
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/12.23.8927
Subject(s) - biology , plasmid , gene , oligonucleotide , yeast , fusion protein , amino acid , fusion gene , coding region , microbiology and biotechnology , mutagenesis , site directed mutagenesis , chimeric gene , dna , peptide sequence , saccharomyces cerevisiae , secretion , biochemistry , gene expression , recombinant dna , mutation , mutant
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.