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Expression of the B subunit of E. coli heat-labile enterotoxin in tobacco using a herbicide resistance gene as a selection marker

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Abstract

The B subunit of Escherichia coli heat-labile enterotoxin (LTB) has been transformed to plants for use as an edible vaccine. We have developed a simple and reliable Agrobacterium-mediated transformation method to express synthetic LTB gene in N. tabacum using a phosphinothricin acetyltransferase (bar) gene as a selectable marker. The synthetic LTB gene adapted to the coding sequence of tobacco plants was cloned to a plant expression vector under the control of the ubiquitin promoter and transformed to tobacco by Agrobacterium-mediated transformation. Transgenic plants were selected in the medium supplemented with 5 mg l-1 phosphinothricin (PPT). The amount of LTB protein detected in the transgenic tobacco was approximately 3.3% of the total soluble protein, approximately 300-fold higher than in the plants generated using the native LTB gene under the control of the CaMV 35S promoter. The transgenic plants that were transferred to a greenhouse had harvested seeds that proved to be resistant to herbicide. Thus, the described protocol could provide a useful tool for the transformation of tobacco plants.

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Abbreviations

CaMV 35S:

cauliflower mosaic virus 35S RNA promoter

GM1:

galactosyl-N-acetylgalactosamyl-sialyl-galactosylglucosyl ceramide

LTB:

B subunit of Escherichia coli heat-labile enterotoxin

PPT:

phosphinothricin

TSP:

total soluble protein

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Authors and Affiliations

  1. Institute of Basic Science, Chonbuk National University, 561-756, Jeonju, South Korea

    Tae-Jin Kang

  2. Division of Biological Sciences and the Research Center for Bioactive Materials, Chonbuk National University, Jeonju, 561-756, South Korea

    So-Chon Han & Moon-Sik Yang

Authors
  1. Tae-Jin Kang
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  2. So-Chon Han
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  3. Moon-Sik Yang
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Correspondence to Moon-Sik Yang.

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Kang, TJ., Han, SC. & Yang, MS. Expression of the B subunit of E. coli heat-labile enterotoxin in tobacco using a herbicide resistance gene as a selection marker. Plant Cell Tiss Organ Cult 81, 165–174 (2005). https://doi.org/10.1007/s11240-004-4734-x

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  • DOI: https://doi.org/10.1007/s11240-004-4734-x

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