Abstract
Transient expression of viral proteins in plants is a novel alternative to other expression platforms. The viral proteins can be used as potential vaccines or in diagnostics. Nicotiana benthamiana leaves or whole plants are infiltrated with recombinant Agrobacterium that harbor the gene of interest. Protein expression in the plants is rapid and results are obtained within 2–7 days. Here we describe how to make electrocompetent Agrobacterium, how to transform Agrobacterium, how to infiltrate leaves or plants with the recombinant Agrobacterium, and lastly how to extract the protein for analysis by gel electrophoresis.
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Acknowledgements
We would like to thank Ed Rybicki, Ann Meyers, and everyone in the Biopharming Research Unit at University of Cape Town for their contribution to the development of the techniques. We acknowledge the National Research Foundation, Department of Science and Technology, the Poliomyelitis Research Foundation, the Medical Research Council, the University of Cape Town Research Committee, the Cancer Research Foundation of South Africa, the PlaProVa project of the EU FP7 initiative, and Era Biotech for funding.
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Hitzeroth, I.I., van Zyl, A.R. (2016). Transient Expression of Viral Proteins in Plants Using Agrobacterium tumefaciens. In: Thomas, S. (eds) Vaccine Design. Methods in Molecular Biology, vol 1404. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-3389-1_38
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DOI: https://doi.org/10.1007/978-1-4939-3389-1_38
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-4939-3389-1
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