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Expression of the B Subunit of E. coli Heat-labile Enterotoxin in the Chloroplasts of Plants and its Characterization

Transgenic Research volume 12, pages 683–691 (2003)Cite this article

Abstract

Transgenic chloroplasts have become attractive systems for heterologous gene expressions because of unique advantages. Here, we report a feasibility study for producing the nontoxic B subunit of Escherichia coli heat-labile enterotoxin (LTB) via chloroplast transformation of tobacco. Stable site-specific integration of the LTB gene into chloroplast genome was confirmed by PCR and genomic Southern blot analysis in transformed plants. Immunoblot analysis indicated that plant-derived LTB protein was oligomeric, and dissociated after boiling. Pentameric LTB molecules were the dominant molecular species in LTB isolated from transgenic tobacco leaf tissues. The amount of LTB protein detected in transplastomic tobacco leaf was approximately 2.5% of the total soluble plant protein, approximately 250-fold higher than in plants generated via nuclear transformation. The GM1–ELISA binding assay indicated that chloroplast-synthesized LTB protein bound to GM1-ganglioside receptors. LTB protein with biochemical properties identical to native LTB protein in the chloroplast of edible plants opens the way for inexpensive, safe, and effective plant-based edible vaccines for humans and animals.

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Affiliations

  1. Basic Sciences Research Institute, Chonbuk National University, Jeonju, 561-756, 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

    Nguyen-Hoang Loc, Mi-Ok Jang, Yong-Suk Jang & Moon-Sik Yang

  3. Institute of Molecular Biology and Genetics, Chonbuk National University, Jeonju, 561-756, South Korea

    Young-Sook Kim

  4. Department of Molecular and Genomic Medicine, College of Medicine, Seoul National University, Seoul, 110-744, sSouth Korea

    Jo-Eun Seo

Authors
  1. Tae-Jin Kang
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  2. Nguyen-Hoang Loc
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  3. Mi-Ok Jang
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  4. Yong-Suk Jang
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  5. Young-Sook Kim
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  6. Jo-Eun Seo
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  7. Moon-Sik Yang
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Correspondence to Moon-Sik Yang.

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Kang, TJ., Loc, NH., Jang, MO. et al. Expression of the B Subunit of E. coli Heat-labile Enterotoxin in the Chloroplasts of Plants and its Characterization. Transgenic Res 12, 683–691 (2003). https://doi.org/10.1023/B:TRAG.0000005114.23991.bc

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  • DOI: https://doi.org/10.1023/B:TRAG.0000005114.23991.bc

  • B subunit of E. coli heat-labile enterotoxin (LTB)
  • chloroplast transformation
  • edible vaccine
  • homologous recombination
  • homoplastomy