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The intergenic region of Arabidopsis thaliana cab1 and cab2 divergent genes functions as a bidirectional promoter

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Abstract

Genetic engineering plays a unique role in fundamental plant biology studies and in improving crop traits. These efforts often necessitate introduction and expression of multiple genes using promoters from a very limited repertoire. Current common practice of expressing multiple genes is the repeated use of the same or similar promoters. This practice causes more frequent transgene silencing due to a high degree of sequence homology and a greater chance of rearrangement among repeatedly used promoter sequences. Therefore, availability and use of natural bidirectional promoters to minimize gene silencing and achieve desirable expression pattern of transgenes is a critical issue in the field of plant genetic engineering. Here we describe the use of a single natural bidirectional promoter to drive the expression of two reporter genes in onion epidermal cells and in transgenic tobacco plants. We show that (1) the promoter drives the simultaneous expression of GUS and GFP reporter genes after transient expression and stable transformation, (2) the transcription is equally strong in both directions, (3) immediate upstream regions in each direction control transcription independently from each other, and (4) the reporter genes are expressed in leaves and stems but not in roots, as expected from the fact that the endogenous promoter controls the expression of two photosynthetic genes in Arabidopsis. Hence, use of bidirectional promoters in heterologous background provides a means to express multiple genes in transgenic plants and aids genetic engineering-based crop improvement.

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Abbreviations

Cab :

Chlorophyll a/b-binding protein

GUS:

β-Glucuronidase

GFP:

Green-fluorescent protein

URE:

Upstream regulatory elements

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Acknowledgments

We thank Amber Davis and Heather Janzen for expert technical assistance. Anish Mitra was an undergraduate summer research intern. This work was supported in part by grants from USDA-NRI and NSF to AM, a contribution of the University of Nebraska Agricultural Research Division.

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Authors and Affiliations

  1. Department of Plant Pathology, University of Nebraska-Lincoln, Lincoln, NE, 68583, USA

    Anish Mitra, Jigang Han & Amitava Mitra

  2. Plant Transformation Core Facility, Division of Plant Sciences, University of Missouri, Columbia, MO, 65211, USA

    Zhanyuan J. Zhang

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  1. Anish Mitra
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  2. Jigang Han
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  3. Zhanyuan J. Zhang
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  4. Amitava Mitra
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Correspondence to Amitava Mitra.

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Mitra, A., Han, J., Zhang, Z.J. et al. The intergenic region of Arabidopsis thaliana cab1 and cab2 divergent genes functions as a bidirectional promoter. Planta 229, 1015–1022 (2009). https://doi.org/10.1007/s00425-008-0859-1

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  • DOI: https://doi.org/10.1007/s00425-008-0859-1

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