A novel yeast secretion vector utilizing secretion signal of killer toxin encoded on the yeast linear DNA plasmid pGKL1.
The NH2-terminal signal region comprising of approximately 70% length of the prepro-sequence of the pGKL killer precursor protein was found to direct an efficient secretion of the mouse alpha-amylase into the culture medium of Saccharomyces cerevisiae. The alpha-amylase molecule secreted into the culture medium was identified by both immuno-blotting and assay of the enzyme activity. The amount of alpha-amylase secreted via the killer toxin signal was comparable to that directed by the leader sequence of mating factor alpha. The secretion of alpha-amylase using the killer toxin signal was blocked at 37C but not at 25C in sec18-1 host, indicating that alpha-amylase is exported through the normal secretion pathway of S. cerevisiae.
D000595:Amino Acid Sequence; D000818:Animals; D005656:Fungal Proteins / Q000235:genetics*; D005796:Genes; D005800:Genes, Fungal; D005822:Genetic Vectors; D055646:Killer Factors, Yeast; D051379:Mice; D009183:Mycotoxins / Q000235:genetics*; D010735:Phosphoglycerate Kinase / Q000235:genetics*; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D000516:alpha-Amylases / Q000235:genetics
D005656:Fungal Proteins; D055646:Killer Factors, Yeast; D009183:Mycotoxins; D029701:Saccharomyces cerevisiae Proteins; D010735:Phosphoglycerate Kinase; D000516:alpha-Amylases