Test systems to study transcriptional regulation and promoter activity in Bacillus megaterium.
Plasmid-located (multi-copy) and chromosomally located (single-copy) promoter test systems were developed for Bacillus megaterium by making use of the homologous beta-galactosidase-encoding bgaM gene. The multi-copy system facilitates rapid promoter analyses and promoter trapping, whereas the single-copy system, integrated into the chromosome, allows investigation of tightly regulated promoters. As a prerequisite for both the multi- and the single-copy systems, a beta-galactosidase-deficient B. megaterium strain was generated by deletion mutagenesis. Both test systems were verified using the promoter of the xylose operon (P( xylA )) from B. megaterium along with its repressor (XylR). As expected, expression levels in the two systems differed significantly, although expression of the bgaM reporter gene was induced by xylose in both cases, thereby proving the functionality of both the multi- and the single-copy system.
Schmidt
Silke
S
Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität Münster, Germany.
Wolf
Nadine
N
Strey
Jan
J
Nahrstedt
Hannes
H
Meinhardt
Friedhelm
F
Waldeck
Jens
J
D001410:Bacillus megaterium / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D005809:Genes, Regulator; D005822:Genetic Vectors; D009876:Operon; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic; D014994:Xylose / Q000378:metabolism*; D001616:beta-Galactosidase / Q000235:genetics* / Q000378:metabolism
D014994:Xylose; D001616:beta-Galactosidase