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Expression of cholera toxin B subunit and the B chain of human insulin as a fusion protein in transgenic tobacco plants

  • Genetic Transformation and Hybridization
  • Published:
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Abstract

A DNA construct containing the cholera toxin B subunit (CTB) gene genetically fused to a nucleotide sequence encoding three copies of tandemly repeated diabetes-associated autoantigen, the B chain of human insulin, was produced and transferred into low-nicotine tobaccos by Agrobacterium. Integration of the fusion gene into the plant genome was confirmed by polymerase chain reaction (PCR). The results of immunoblot analysis verified the synthesis and assembly of the fusion protein into pentamers in transgenic tobacco. GM1–ELISA showed that the plant-derived fusion protein retained GM1–ganglioside receptor binding specificity. The fusion protein accounted for 0.11% of the total leaf protein. The production of transgenic plants expressing CTB–InsB3 offers a new opportunity to test plant-based oral antigen therapy against autoimmune diabetes by inducing oral tolerance.

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Acknowledgements

The authors thank Dr. J. Holmgren, Institute of Medical Microbiology, University of Göteborg, Sweden, for providing bacterial CTB DNA clone. This research was supported by the Natural Sciences and Engineering Research Council (NSERC) and the London Health Sciences Centre MOTS Research Fund.

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Authors and Affiliations

  1. Department of Biology, University of Western Ontario, London, Ontario, Canada, N6A 5B7

    Dora Li, Jennifer O’Leary, Norman P. A. Huner & Shengwu Ma

  2. Plantigen Inc., 700 Collip Circle, London, Ontario, Canada, N6G 4X8

    Yan Huang, Anthony M. Jevnikar & Shengwu Ma

  3. Transplantation Immunology Group, Lawson Health Research Institute, London, Ontario, Canada, N6A 4G5

    Anthony M. Jevnikar & Shengwu Ma

Authors
  1. Dora Li
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  2. Jennifer O’Leary
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  3. Yan Huang
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  4. Norman P. A. Huner
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  5. Anthony M. Jevnikar
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  6. Shengwu Ma
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Corresponding author

Correspondence to Shengwu Ma.

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Communicated by H. Ebinuma

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Li, D., O’Leary, J., Huang, Y. et al. Expression of cholera toxin B subunit and the B chain of human insulin as a fusion protein in transgenic tobacco plants. Plant Cell Rep 25, 417–424 (2006). https://doi.org/10.1007/s00299-005-0069-2

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  • DOI: https://doi.org/10.1007/s00299-005-0069-2

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