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Expression of the B subunit of Escherichia coli heat-labile enterotoxin as a fusion protein in transgenic tomato

  • Genetic Transformation and Hybridization
  • Published:
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Abstract

Epitopes often require co-delivery with an adjuvant or targeting protein to enable recognition by the immune system. This paper reports the ability of transgenic tomato plants to express a fusion protein consisting of the B subunit of the Escherichia coli heat-labile enterotoxin (LTB) and an immunocontraceptive epitope. The fusion protein was found to assemble into pentamers, as evidenced by its ability to bind to gangliosides, and had an average expression level of 37.8 μg g−1 in freeze-dried transgenic tissues. Processing of selected transgenic fruit resulted in a 16-fold increase in concentration of the antigen with minimal loss in detectable antigen. The species-specific nature of this epitope was shown by the inability of antibodies raised against non-target species to detect the LTB fusion protein. The immunocontraceptive ability of this vaccine will be tested in future pilot mice studies.

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Fig. 5A, B.

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Abbreviations

DW :

Dry weight

ELISA :

Enzyme-linked immunosorbant assay

ETEC :

Entertoxigenic Escherichia coli

FW :

Fresh weight

LTB :

B subunit of Escherichia coli heat-labile enterotoxin

ZP3 :

Zona pellucida glycoprotein 3

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Acknowledgements

For valuable technical assistance, the investigators would like to thank Que Quiang, Tracey Cranston and Joan Lenz. This work was supported by the Co-operative Research Centre for Conservation and Management of Marsupials, Axis Genetics, and Dow AgroSciences LLC.

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Authors and Affiliations

  1. Boyce Thompson Institute for Plant Research, Cornell University, Ithaca, NY 14850, USA

    A. M. Walmsley, D. D. Kirk & H. S. Mason

  2. Department of Plant Biology, Arizona State University, Tempe, AZ 85287-1601, USA

    A. M. Walmsley, M. L. Alvarez, Y. Jin, J. Pinkhasov, M. M. Rigano & C. J. Arntzen

  3. Department of Primary Industries, Queensland Agricultural Biotechnology Centre, 4072, Brisbane, Queensland, Australia

    S. M. Lee

Authors
  1. A. M. Walmsley
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  2. M. L. Alvarez
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  3. Y. Jin
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  4. D. D. Kirk
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  5. S. M. Lee
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  6. J. Pinkhasov
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  7. M. M. Rigano
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  8. C. J. Arntzen
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  9. H. S. Mason
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Correspondence to A. M. Walmsley.

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Communicated by E.D. Earle

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Walmsley, A.M., Alvarez, M.L., Jin, Y. et al. Expression of the B subunit of Escherichia coli heat-labile enterotoxin as a fusion protein in transgenic tomato. Plant Cell Rep 21, 1020–1026 (2003). https://doi.org/10.1007/s00299-003-0619-4

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  • DOI: https://doi.org/10.1007/s00299-003-0619-4

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