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Isolation and characterization of putative Pseudobutyrivibrio ruminis promoters
Journal article   Peer reviewed

Isolation and characterization of putative Pseudobutyrivibrio ruminis promoters

T.D. Schoep and K. Gregg
Microbiology, Vol.153(9), pp.3071-3080
09/2007
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Abstract

Novel plasmids were constructed for the analysis of DNA fragments from the rumen bacterium Pseudobutyrivibrio ruminis. Five previously unidentified promoters were characterized using a novel primer extension method to identify transcription start sites. The genes downstream of these promoters were not identified, and their activity in expression of genomic traits in wild-type P. ruminis remains putative. Comparison with promoters from this and closely related species revealed a consensus sequence resembling the binding motif for the RNA polymerase σ 70-like factor complex. Consensus -35 and -10 sequences within these elements were TTGACA and ATAATATA respectively, interspaced by 15-16 bp. The consensus for the -10 element was extended by one nucleotide upstream and downstream of the standard hexamer (indicated in bold). Promoter strengths were measured by reverse transcription quantitative PCR and β-glucuronidase assays. No correlation was found between the composition and context of elements within P. ruminis promoters, and promoter strength. However, a mutation within the -35 element of one promoter revealed that transcriptional strength and choice of transcription start site were sensitive to this single nucleotide change.

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Collaboration types
Domestic collaboration
Citation topics
1 Clinical & Life Sciences
1.42 Bacteriology
1.42.131 Bacterial Gene Regulation
Web Of Science research areas
Microbiology
ESI research areas
Microbiology
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