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Expression and Assembly of Cholera Toxin B Subunit (CTB) in Transgenic Carrot (Daucus carota L.)

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Abstract

We expressed the cholera toxin B subunit (CTB) fused to an endoplasmic reticulum retention signal (SEKDEL) in carrot roots using an Agrobacterium-mediated transformation method. Fourteen independent transgenic lines were regenerated via somatic embryogenesis after 6 months of culture. The sCTB gene was detected in the genomic DNA of transgenic carrot by PCR amplification. Expressions and assembly of sCTB protein into oligomeric structures of pentameric size were observed in transgenic plant extracts by Western blot analysis. The sCTB produced by transgenic carrot roots demonstrated strong affinity for GM1-ganglioside, suggesting that the sCTB conserved the antigenic sites for binding and proper folding of the pentameric sCTB structure. The expression level of sCTB comprised approximately 0.48% of total soluble protein (TSP) in root of transgenic carrot.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic acid

GM1 :

Galactosyl-N-acetylgalactosamylsialyl-galactosylglucosylceramide

sCTB:

Synthetic cholera toxin B subunit

TSP:

Total soluble protein

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Acknowledgements

This work was supported by a Korea Research Foundation (KRF-2006-0005-J03103).

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Authors and Affiliations

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

    Young-Sook Kim, Mi-Young Kim, Tae-Geum Kim & Moon-Sik Yang

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Correspondence to Moon-Sik Yang.

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Kim, YS., Kim, MY., Kim, TG. et al. Expression and Assembly of Cholera Toxin B Subunit (CTB) in Transgenic Carrot (Daucus carota L.). Mol Biotechnol 41, 8–14 (2009). https://doi.org/10.1007/s12033-008-9086-z

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