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Expression of Escherichia coli heat-labile enterotoxin b subunit (LTB) in carrot (Daucus carota L.)

Plant Cell Reports volume 26, pages 969–976 (2007)Cite this article

Abstract

We expressed the B subunit of enterotoxigenic Escherichia coli heat-labile enterotoxin (LTB) encoded by a synthetic codon-optimized gene in carrot. An Agrobacterium-mediated transformation method was used. Thirty independent transgenic lines were regenerated via somatic embryogenesis after 6 months in culture and were transferred to a greenhouse. GM1-ELISA assay was used to assess LTB protein content in mature taproots. Some transgenic lines expressed LTB up to 0.3% of the total soluble protein, which is tenfold higher than the expression levels reported earlier using the native bacterial gene in plants. Immunological assay confirmed proper assembly of the pentameric complex and in vitro activity of the recombinant LTB protein, suggesting that it can be functional in prevention of diarrhea.

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Abbreviations

CaMV 35S:

Cauliflower mosaic virus 35S RNA promoter

GM1:

Galactosyl-N-acetylgalactosamyl-sialyl-galactosylglucosyl ceramide

LTB:

B subunit of the heat-labile enterotoxin of Escherichia coli

TSP:

Total soluble protein

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Acknowledgments

This research was funded by grant 37048-B and the scholarships 165946 and 172307 for the graduate studies of Sergio Rosales-Mendoza and Ruth Soria-Guerra, respectively, from CONACYT México, and by an UC-MexUS grant to Rubén López-Revilla. Thanks are due to José Luis Sánchez for suggesting the use of carrots and Dr. John Clements for the LTB donation.

Author information

Affiliations

  1. División de Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica, Camino a la Presa San José 2055, 78216, San Luis Potosí, S.L.P., Mexico

    Sergio Rosales-Mendoza, Ruth Elena Soria-Guerra, Rubén López-Revilla, Gerardo Rafael Argüello-Astorga, Juan Francisco Jiménez-Bremont & Ángel Gabriel Alpuche-Solís

  2. Facultad de Química, Universidad Nacional Autónoma de México, Ciudad Universitaria, 04510, México, D.F., Mexico

    María Teresa de Jesús Olivera-Flores

  3. Facultad de Ciencias Químicas, Universidad Autónoma de San Luis Potosí, Av. Dr. Manuel Nava No. 6, 78210, San Luis Potosí, S.L.P., Mexico

    Ramón Fernando García-de la Cruz

  4. Maestría en Ciencias Odontológicas, Universidad Autónoma de San Luis Potosí, Av. Dr. Manuel Nava No. 2, 78290, San Luis Potosí, S.L.P., Mexico

    Juan Pablo Loyola-Rodríguez

Authors
  1. Sergio Rosales-Mendoza
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  2. Ruth Elena Soria-Guerra
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  3. María Teresa de Jesús Olivera-Flores
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  4. Rubén López-Revilla
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  5. Gerardo Rafael Argüello-Astorga
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  6. Juan Francisco Jiménez-Bremont
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  7. Ramón Fernando García-de la Cruz
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  8. Juan Pablo Loyola-Rodríguez
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  9. Ángel Gabriel Alpuche-Solís
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Corresponding author

Correspondence to Ángel Gabriel Alpuche-Solís.

Additional information

Communicated by P. Lakshmanan.

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Rosales-Mendoza, S., Soria-Guerra, R.E., de Jesús Olivera-Flores, M.T. et al. Expression of Escherichia coli heat-labile enterotoxin b subunit (LTB) in carrot (Daucus carota L.). Plant Cell Rep 26, 969–976 (2007). https://doi.org/10.1007/s00299-007-0310-2

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  • DOI: https://doi.org/10.1007/s00299-007-0310-2

Keywords

  • Diarrhea prevention
  • Plant based-vaccine
  • Synthetic gene
  • Expression levels
  • Somatic embryogenesis