title: Secretion of a heterologous protein from Bacillus subtilis with the aid of protease signal sequences.
abstract: Secretion vectors based on the genes from Bacillus amyloliquefaciens P for alkaline protease (aprBamP) and neutral protease (nprBamP) were constructed. With both aprBamP and nprBamP, a unique restriction site was introduced 3' of the predicted signal coding region by using the technique of oligonucleotide-directed mutagenesis. The new sites enabled us to fuse a heterologous gene to the expression and secretion elements. We used the protein A gene (spa) from Staphylococcus aureus as a heterologous gene. Bacillus subtilis cells carrying the resulting apr-spa or npr-spa gene fusions synthesized the fusion protein. B. subtilis cells were also capable of removing the signal peptide from the fusion protein, as indicated by the appearance of processed protein A into the growth medium. In addition, these gene fusions allowed us to identify the signal processing site of both the APR-SPA and NPR-SPA proteins.
authors.forename: N
authors.initials: N
authors.lastname: Vasantha
authors.forename: L D
authors.initials: LD
authors.lastname: Thompson
mesh_terms: D001407:Bacillus / Q000201:enzymology; D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D004274:DNA, Recombinant; D010450:Endopeptidases / Q000235:genetics* / Q000378:metabolism*; D005822:Genetic Vectors; D009154:Mutation; D015260:Neprilysin; D021382:Protein Sorting Signals / Q000235:genetics / Q000378:metabolism*; D012697:Serine Endopeptidases; D013205:Staphylococcal Protein A / Q000235:genetics / Q000378:metabolism*
chemical_list: D004274:DNA, Recombinant; D021382:Protein Sorting Signals; D013205:Staphylococcal Protein A; D010450:Endopeptidases; D012697:Serine Endopeptidases; C032923:microbial serine proteinases; D015260:Neprilysin