Expression of the neutral protease gene from a thermophilic Bacillus sp. BT1 strain in Bacillus subtilis and its natural host: identification of a functional promoter.
The expression of the neutral protease gene (npr) from the thermophilic Bacillus sp. BT1 strain was studied in its natural host and in mesophilic Bacillus subtilis. In the thermophilic BT1 strain, the transcription of the protease gene is initiated from its own promoter, just 5' to the gene. In contrast, in heterologous B. subtilis this thermophilic npr promoter does not function, and expression of the npr gene results from transcription originating upstream of an adjacent gene, open reading frame X (ORF X). A functional promoter was identified 5' to ORF X that is required for efficient expression of the npr gene in Bacillus subtilis as verified by primer extension, reverse transcription-PCR, and 5' rapid amplification of cDNA ends experiments. These data suggest that transcriptional signals used in thermophilic Bacillus sp. BT1 strain are different from those used in B. subtilis.
Vecerek
B
B
Laboratory of Enzyme Technology, Institute of Microbiology CAS, Prague, Czech Republic. vecerek@bimed.cas.cz
Venema
G
G
D001407:Bacillus / Q000201:enzymology / Q000235:genetics*; D001412:Bacillus subtilis; D001483:Base Sequence; D005784:Gene Amplification; D015964:Gene Expression Regulation, Bacterial; D006358:Hot Temperature; D008666:Metalloendopeptidases / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011401:Promoter Regions, Genetic; D017422:Sequence Analysis, DNA
D008666:Metalloendopeptidases; C085836:Npr protease