Synthesis and assembly of Escherichia coli heat-labile enterotoxin B subunit in transgenic lettuce (Lactuca sativa)

Tae Geum Kim, Mi Young Kim, Bang Geul Kim, Tae Jin Kang, Young Sook Kim, Yong Suk Jang, Charles J. Arntzen, Moon Sik Yang

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Abstract

Escherichia coli heat-labile enterotoxin B subunit (LTB) strongly induces immune responses and can be used as an adjuvant for co-administered antigens. Synthetic LTB (sLTB) based on optimal codon usage by plants was introduced into lettuce cells (Lactuca sativa) by Agrobacterium tumefaciens-mediated transformation methods. The sLTB gene was detected in the genomic DNA of transgenic lettuce leaf cells by PCR DNA amplification. Synthesis and assembly of the sLTB protein into oligomeric structures of pentameric size was observed in transgenic plant extracts using Western blot analysis. The binding of sLTB pentamers to intestinal epithelial cell membrane glycolipid receptors was confirmed by GM1-ganglioside enzyme-linked immunosorbent assay (GM1-ELISA). Based on the results of ELISA, sLTB protein comprised approximately 1.0-2.0% of total soluble protein in transgenic lettuce leaf tissues. The synthesis and assembly of sLTB monomers into biologically active oligomers in transgenic lettuce leaf tissues demonstrates the feasibility of the use of edible plant-based vaccines consumed in the form of raw plant materials to induce mucosal immunity.

Original languageEnglish (US)
Pages (from-to)22-27
Number of pages6
JournalProtein Expression and Purification
Volume51
Issue number1
DOIs
StatePublished - Jan 1 2007

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Keywords

  • Edible vaccine
  • Escherichia coli heat-labile enterotoxin B subunit
  • Lettuce
  • Mucosal immunization

ASJC Scopus subject areas

  • Biotechnology

Cite this

Kim, T. G., Kim, M. Y., Kim, B. G., Kang, T. J., Kim, Y. S., Jang, Y. S., Arntzen, C. J., & Yang, M. S. (2007). Synthesis and assembly of Escherichia coli heat-labile enterotoxin B subunit in transgenic lettuce (Lactuca sativa). Protein Expression and Purification, 51(1), 22-27. https://doi.org/10.1016/j.pep.2006.05.024