A novel Bacillus subtilis expression vector based on bacteriophage phi 105.
We have developed a novel expression vector based on the bacteriophage phi 105, and employed it for the production of mutant beta-lactamases in Bacillus subtilis. Expression of the beta-lactamase-encoding gene was low when cloned into the prophage under the control of its own promoter. However, expression was considerably elevated when the gene was inserted into the phage genome in the same orientation as phage transcription. A defective phi 105 vector was constructed with a deletion removing a region needed for cell lysis, and with a mutation in the immunity repressor, rendering it temperature sensitive. Production of beta-lactamase could then be induced by a shift in temperature and without concomitant cell lysis, facilitating purification of the protein from the culture supernatant. This phage has considerable potential for development as a vector for controllable production of heterologous proteins in B. subtilis.
Gibson
R M
RM
Sir William Dunn School of Pathology, University of Oxford, UK.
Errington
J
J
D001412:Bacillus subtilis / Q000235:genetics*; D001435:Bacteriophages / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D005798:Genes, Bacterial; D005822:Genetic Vectors; D009154:Mutation; D011401:Promoter Regions, Genetic; D001618:beta-Lactamases / Q000096:biosynthesis / Q000235:genetics
D001618:beta-Lactamases