» Articles » PMID: 10869092

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

Overview
Journal J Bacteriol
Specialty Microbiology
Date 2000 Jun 27
PMID 10869092
Authors
Affiliations
Soon will be listed here.
Abstract

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.

References
1.
Moran Jr C, Lang N, LeGrice S, Lee G, Stephens M, Sonenshein A . Nucleotide sequences that signal the initiation of transcription and translation in Bacillus subtilis. Mol Gen Genet. 1982; 186(3):339-46. DOI: 10.1007/BF00729452. View

2.
Doi R . Genetic engineering in Bacillus subtilis. Biotechnol Genet Eng Rev. 1984; 2:121-55. DOI: 10.1080/02648725.1984.10647797. View

3.
Chavira Jr R, BURNETT T, Hageman J . Assaying proteinases with azocoll. Anal Biochem. 1984; 136(2):446-50. DOI: 10.1016/0003-2697(84)90242-2. View

4.
Eijsink V, Veltman O, Aukema W, Vriend G, Venema G . Structural determinants of the stability of thermolysin-like proteinases. Nat Struct Biol. 1995; 2(5):374-9. DOI: 10.1038/nsb0595-374. View

5.
Imanaka T, Shibazaki M, Takagi M . A new way of enhancing the thermostability of proteases. Nature. 1986; 324(6098):695-7. DOI: 10.1038/324695a0. View