caa a22 4500 475745809 CHVBK 20180406123513.0 cr unu---uuuuu 170329e20000301xx s 000 0 eng 10.1007/s002849910033 doi (NATIONALLICENCE)springer-10.1007/s002849910033 Construction of a Promoter-Rescue Plasmid for Butyrivibrio fibrisolvens and Its Use in Characterization of a Flagellin Promoter [Elektronische Daten] [Cheryl E. Beard, Keith Gregg, Martin Kalmokoff, Ron M. Teather] 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. Springer-Verlag New York Inc., 2000 Beard Cheryl E. CSIRO Division of Entomology, GPO Box 1700, Canberra, A.C.T., 2614, Australia, AU aut Gregg Keith Biological Sciences and Biotechnology, Murdoch University, Perth, W.A. 6150, Australia, AU aut Kalmokoff Martin Microbiology Research Division, Health Canada, Banting Research Centre, Postal Locator 2204A2, Tunney's Pasture, Ottawa, Ontario, K1A 0L2, Canada, CA aut Teather Ron M. Agriculture Canada, Lethbridge Research Centre, PO Box 3000, Lethbridge, ABT1J 4B1, Canada, CA aut https://doi.org/10.1007/s002849910033 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s002849910033 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Beard Cheryl E. CSIRO Division of Entomology, GPO Box 1700, Canberra, A.C.T., 2614, Australia, AU aut NATIONALLICENCE 700 1- Gregg Keith Biological Sciences and Biotechnology, Murdoch University, Perth, W.A. 6150, Australia, AU aut NATIONALLICENCE 700 1- Kalmokoff Martin Microbiology Research Division, Health Canada, Banting Research Centre, Postal Locator 2204A2, Tunney's Pasture, Ottawa, Ontario, K1A 0L2, Canada, CA aut NATIONALLICENCE 700 1- Teather Ron M. Agriculture Canada, Lethbridge Research Centre, PO Box 3000, Lethbridge, ABT1J 4B1, Canada, CA aut Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer