Functional analysis of a nitrite reductase promoter from birch in transgenic tobacco.
Nitrate assimilation is a highly regulated process in higher plants, and the regulatory cues governing gene expression in this pathway include both external and internal factors. In birch (Betula pendula Roth) the expression of nitrate reductase (NR) and nitrite reductase (NiR) genes is co-regulated by light and nitrate at the transcriptional level. In order to identify cis-acting DNA-elements involved in light and nitrate induction of the birch NiR gene, a 0.9 kb 5' flanking region of the NiR gene was isolated, analysed on the DNA level, and the transcription start site was determined. Deletion analysis of the birch NiR promoter region fused to the GUS reporter gene (uidA) in transgenic tobacco (Nicotiana tabacum) revealed the presence of light- and nitrate-responsive promoter fragments. The responsive fragments showed different activities in leaves and roots. Further, gel mobility shift assays using nuclear proteins from leaves detected a specific DNA-binding activity to the sequence between -146 and -267 bp that was induced in darkness and disappeared in the light. The deletion analysis has shown that this region is critical for light inducibility of the birch NiR gene in leaves.
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Botanisches Institut, Universität Bonn, Karlrobert-Kreiten-Str. 13, D-53115, Bonn, Germany
Hachtel
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