Abstract.
The Butyrivibrio fibrisolvens/Escherichia coli shuttle vector pBHerm has been modified to produce a plasmid (pBHE) that can be used for the identification and characterization of promoters in B. fibrisolvens. pBHE allows the insertion of a test promoter immediately upstream of a promoterless erythromycin resistance gene (ermAM). The efficacy of the pBHE plasmid in isolating and characterizing promoters was tested by inserting the flagellin gene (flaA) promoter from B. fibrisolvens OR77. Transcription of the ermAM gene from the flaA promoter was significantly higher than that observed when the ermAM gene was under the control of its own promoter. The flagelling gene of OR77 appears to be transcribed from two different promoters that produce transcripts initiating approximately 130 bp apart. Two mutant flaA promoter constructs, containing mutations in the −10 and −35 regions of either of the two putative promoter regions, showed drastic alterations in both the origin and amounts of the two transcripts produced. Mutations in either promoter affected transcription from both promoters, indicating that both regions contribute to gene expression.
Similar content being viewed by others
Author information
Authors and Affiliations
Additional information
Received: 26 July 1999 / Accepted: 14 October 1999
Rights and permissions
About this article
Cite this article
Beard, C., Gregg, K., Kalmokoff, M. et al. Construction of a Promoter-Rescue Plasmid for Butyrivibrio fibrisolvens and Its Use in Characterization of a Flagellin Promoter. Curr Microbiol 40, 164–168 (2000). https://doi.org/10.1007/s002849910033
Issue Date:
DOI: https://doi.org/10.1007/s002849910033