2000 Congress Symposium on Molecular Farming

In Vitro Cellular & Developmental Biology - Plant

, Volume 38, Issue 1, pp 11-17

Development of an edible subunit vaccine in corn against enterotoxigenic strains of escherichia coli

  • Stephen J. StreatfieldAffiliated withProdiGene Email author 
  • , Jocelyne M. MayorAffiliated withProdiGene
  • , Donna K. BarkerAffiliated withProdiGene
  • , Christopher BrooksAffiliated withProdiGene
  • , Barry J. LamphearAffiliated withProdiGene
  • , Susan L. WoodardAffiliated withProdiGene
  • , Katherine K. BeifussAffiliated withProdiGene
  • , Debra V. VicunaAffiliated withProdiGene
  • , Leigh Anne MasseyAffiliated withProdiGene
    • , Michael E. HornAffiliated withProdiGene
    • , Donna E. DelaneyAffiliated withProdiGene
    • , Zivko L. NikolovAffiliated withProdiGene
    • , Elizabeth E. HoodAffiliated withProdiGene
    • , Joseph M. JilkaAffiliated withProdiGene
    • , John A. HowardAffiliated withProdiGene

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Advances in the development of subunit vaccines and in the production of foreign proteins in plants together offer the prospect of stable and inexpensive vaccine delivery systems. Various bacterial and viral proteins stably produced in plants have been shown to elicit immune responses in feeding trials. We have extended this approach by using Zea mays as the plant production system. Corn has several advantages as a vaccine delivery vehicle, most notably established technologies to generate transgenic plants, to optimize traits through breeding and to process the seed into a palatable form. Here we report on the production in corn seed of the GM1 receptor binding (B) subunit of the heat-labile toxin (Lt) from enterotoxigenic strains of Escherichia coli. Versions of the Lt-B gene were synthesized to give optimum codon usage for corn and to target the protein to either the cell surface or the cytoplasm. These synthetic genes were fused to a strong promoter and transformed into corn. Lt-B was highly expressed in corn seed at up to 1.8% of the total soluble protein and this was further increased approximately five-fold through plant breeding. As in E. coli. Lt-B produced in corn forms a functional pentamer that can bind to the GM1 receptor. Furthermore, Lt-B pentamer stored in corn seed is much more resistant to heat than is the pure protein, allowing the transgenic corn to be readily processed into an edible form. This work demonstrates the potential of using products derived from transgenic corn seed as delivery vehicles for subunit vaccines.

Key words

enterotoxigenic Escherichia coli heat-labile toxin protein stabilization subcellular targeting transgene expression transgenic corn