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PMID: 3081490 Published · ppublish English Journal Article

Secretion of a heterologous protein from Bacillus subtilis with the aid of protease signal sequences.

Journal of bacteriology ·Vol. 165 ·No. 3 ·1986-03-00 ·Pages 837-42

Vasantha N, Thompson LD

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.

MeSH Terms
Bacillus/enzymology Bacillus subtilis/enzymology,genetics,metabolism DNA, Recombinant Endopeptidases/genetics,metabolism Genetic Vectors Mutation Neprilysin Protein Sorting Signals/genetics,metabolism Serine Endopeptidases Staphylococcal Protein A/genetics,metabolism
Chemicals
DNA, Recombinant Protein Sorting Signals Staphylococcal Protein A Endopeptidases Serine Endopeptidases microbial serine proteinases Neprilysin
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Vasantha N
Thompson L D
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.165.3.837-842.1986
Published
1986-03-00
Pages
837-42
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC214504
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.165.3.837-842.1986 PMC Full Text View journal details
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