Secretion of staphylococcal nuclease by Bacillus subtilis.
The staphylococcal nuclease (nuc) gene from Staphylococcus aureus has been cloned and expressed in Bacillus subtilis. The nuclease protein was expressed either from its own promoter and translation start signals, or from a combination of a B. subtilis promoter, ribosome binding site, and a signal peptide sequence. Greater than 80% of the active gene product was secreted into the medium, whereas, when a signal peptide sequence was absent, as little as 4% of the nuclease activity was found in the culture medium. Intracellular (or cell-bound) nuclease, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, was shown to have the molecular weight of the predicted precursor protein with the signal peptide. Levels of nuclease reached 50 mg per liter in the culture medium, depending on the growth medium and the strain used. These findings indicate the prospective use of nuclease as a model system for studying secretion of heterologous proteins in B. subtilis.
D001412:Bacillus subtilis / Q000235:genetics*; D002118:Calcium / Q000378:metabolism; D003001:Cloning, Molecular; D003062:Codon; D004591:Electrophoresis, Polyacrylamide Gel; D007163:Immunosorbent Techniques; D008836:Micrococcal Nuclease / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D010455:Peptides / Q000378:metabolism; D014176:Protein Biosynthesis; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals; D014158:Transcription, Genetic
D003062:Codon; D010455:Peptides; D021382:Protein Sorting Signals; D008836:Micrococcal Nuclease; D002118:Calcium