In vitro characterization of the ORF1-ntrBC promoter of Rhizobium etli.

Rhizobium sigma vegetative-dependent promoters are different from those of enteric bacteria and have never been characterized before. We report here the biochemical characterization of the ORF1-ntrBC promoter of Rhizobium etli. The minimal promoter region was located by means of a transcriptional fusion and further characterized by in vitro transcription and gel retardation experiments. Oligonucleotides used as DNA competitors in runoff transcription experiments allowed the precise localisation of the promoter region. Protein extracts from an ntrC+, but not from an ntrC- strain, inhibited in vitro transcription. The NtrC protein was found to bind specifically to the promoter, where an NtrC binding site overlapping the transcription initiation site, is present.