Establishment of a simple and rapid method to screen for strong promoters in Bacillus subtilis.
Using published plasmid vectors containing the bgaB gene encoding a heat-stable beta-galactosidase, we have been unable to fuse strong promoters to this reporter gene. In addition, we could not analyze the promoter strength by a plate assay. Therefore, we constructed an Escherichia coli-Bacillus subtilis shuttle vector to allow the cloning of strong promoters and their rapid analysis in B. subtilis by plating on X-Gal plates in the presence of the inducer IPTG. We show that the blue color of the colonies reflects the strength of the promoters, which was verified by measuring the beta-galactosidase activities.
Phan
Trang Thi Phuong
TT
Chemical Genomics Centre, Max-Planck Institute of Molecular Physiology, Otto-Hahn-Str. 15, Dortmund, Germany.
Nguyen
Hoang Duc
HD
Schumann
Wolfgang
W
D001412:Bacillus subtilis / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005697:Galactosides / Q000235:genetics* / Q000378:metabolism; D017930:Genes, Reporter / Q000235:genetics*; D005822:Genetic Vectors; D006358:Hot Temperature; D007211:Indoles / Q000378:metabolism; D007544:Isopropyl Thiogalactoside / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics* / Q000378:metabolism
D005697:Galactosides; D007211:Indoles; D007544:Isopropyl Thiogalactoside; D001616:beta-Galactosidase; C044888:5-bromo-4-chloro-3-indolyl beta-galactoside