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Localization of light-inducible and tissue-specific regions of the spinach ribulose bisphosphate carboxylase/oxygenase (rubisco) activase promoter in transgenic tobacco plants

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Abstract

Deletions in the spinach rubisco activase (Rca) promoter in transgenic tobacco were analyzed to define the regions necessary for conferring light-inducible and tissue-specific expression. Transgenic plants were constructed with Bal 31 deletions of the Rca promoter fused to the coding region of the bacterial reporter gene β-glucuronidase (GUS). Analysis of the Rca deletion mutants localized the region conferring normal expression downstream from −294 relative to the Rca transcription start site. A second set of transgenic plants containing the cauliflower mosaic virus (CaMV) 35S enhancer fused to the 3′ end of the Rca/GUS constructs demonstrated the presence of a light-responsive element between −150 and −78 active in leaves. Regions 10 bp long within the light-responsive region, which included putative G box and GT elements, were removed by recombinant polymerase chain reaction. Deletion of the G box element resulted in a loss of gene expression in the leaves of transgenic tobacco, while deletion of the GT motif caused a 10–100-fold increase in expression in roots. However, site-directed mutagenesis of the GT motif resulted in expression patterns identical to the normal promoter. These experiments demonstrated that light-inducible and tissue-specific expression of the Rca promoter involves multiple cis elements proximal to the transcription start site, and that interactions between these elements are essential for regulating expression.

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Authors and Affiliations

  1. Department of Plant Biology, University of Illinois at Urbana-Champaign, 61801-3838, Urbana, Illinois, USA

    Beverly M. Orozco & William L. Ogren

  2. Department of Agronomy, University of Illinois at Urbana-Champaign, 61801-3838, Urbana, Illinois, USA

    William L. Ogren

  3. Department of Photosynthesis Research Unit, Agricultural Research Service, United States Department of Agriculture, 61801-3838, Urbana, Illinois, USA

    William L. Ogren

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  1. Beverly M. Orozco
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  2. William L. Ogren
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Orozco, B.M., Ogren, W.L. Localization of light-inducible and tissue-specific regions of the spinach ribulose bisphosphate carboxylase/oxygenase (rubisco) activase promoter in transgenic tobacco plants. Plant Mol Biol 23, 1129–1138 (1993). https://doi.org/10.1007/BF00042347

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  • DOI: https://doi.org/10.1007/BF00042347

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