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Enhanced expression of B-subunit of Escherichia coli heat-labile enterotoxin in tobacco by optimization of coding sequence

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Abstract

Escherichia coli heat-labile toxin (LT) is a potent mucosal immunogen and immunoadjuvant for coadministered antigens. We synthesized a gene encoding the B-subunit of LT(LTB) adapted to the coding sequence of tobacco plants and fused to the endoplasmic reticulum retention signal SEKDEL to enhance its level of expression in plants. The synthetic LTB gene was cloned into a plant expression vector adjacent to the CaMV 35S promoter and was introduced into tobacco by Agrobacterium-mediated transformation. The amount of LTB protein detected in transgenic tobacco leaves was 2.2% of the total soluble plant protein, which is approx 200-fold higher than in previous reports of native LTB gene expression in transgenic plants. Enzyme-linked immunosorbent assay indicated that plant-synthesized LTB protein bound specifically to GM1-ganglioside, suggesting that the LTB subunits formed active pentamers.

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Authors and Affiliations

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

    Tae-Jin Kang

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

    So-Chon Han, Mi-Ok Jang, Kui-Hyeon Kang, Yong-Suk Jang & Moon-Sik Yang

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  1. Tae-Jin Kang
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  2. So-Chon Han
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  3. Mi-Ok Jang
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  4. Kui-Hyeon Kang
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  6. Moon-Sik Yang
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Correspondence to Moon-Sik Yang.

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Kang, TJ., Han, SC., Jang, MO. et al. Enhanced expression of B-subunit of Escherichia coli heat-labile enterotoxin in tobacco by optimization of coding sequence. Appl Biochem Biotechnol 117, 175–187 (2004). https://doi.org/10.1385/ABAB:117:3:175

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  • DOI: https://doi.org/10.1385/ABAB:117:3:175

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