Vectors using the phospho-alpha-(1,1)-glucosidase-encoding gene treA of Bacillus subtilis as a reporter.
The intracellular phospho-alpha-(1,1)-glucosidase, TreA, from Bacillus subtilis (Bs) hydrolyses trehalose 6-phosphate into glucose and glucose 6-phosphate. The enzyme is also able to cleave p-nitrophenyl alpha-D-glucopyranoside (PNPG). This enzymatic reaction can be easily monitored in a beta-galactosidase analogous enzyme assay. The vectors we have constructed can be used to study promoter activity in transcriptional treA fusions and may prove especially useful under high-salt conditions due to the halophilic character of TreA. The treA gene is useful as a reporter in either Bs or Escherichia coli (Ec). Such fusions can be integrated in the Bs amyE locus and selected on either kanamycin or chloramphenicol, or used as plasmids in Ec. As an example of the general utility, we demonstrate treA expression under xylA-operator-promoter control.
Schöck
F
F
Lehrstuhl für Mikrobiologie, Institut für Mikrobiologie, Biochemie und Genetik, Friedrich-Alexander Universität Erlangen-Nürnberg, Germany.
Gotsche
S
S
Dahl
M K
MK
D018528:ATP-Binding Cassette Transporters / Q000235:genetics; D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D002702:Chloramphenicol Resistance; D004186:Disaccharidases / Q000235:genetics* / Q000378:metabolism; D005798:Genes, Bacterial; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics*; D007613:Kanamycin Resistance; D009598:Nitrophenylgalactosides / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012641:Selection, Genetic
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D009598:Nitrophenylgalactosides; C059200:4-nitrophenylgalactoside; D004186:Disaccharidases; C040631:alpha, alpha-phosphotrehalase