Abstract
The production of biopharmaceuticals in plants is currently one of the most attractive approaches to modern medicine. Several efficient plant-based expression systems have been developed so far. Among them, plastid transformation has attracted biotechnologists because the plastid genome, unlike nuclear genome, bears a number of unique advantages for plant genetic engineering. These include higher levels of protein production, uniform gene expression of transformants due to the lack of epigenetic interference, and expression of multiple genes (as in operons) from the same construct. Further, the plastid transformation technology is an environmentally friendly method because plastid and their genetic information are maternally inherited in many species with a consequent lack of transmission of plastid DNA by pollen. Recently, great progress has been made with plastid-based production of biopharmaceuticals demonstrating that it is a promising platform for such purposes. This chapter describes detailed protocols for plastid transformation including the delivery of DNA by biolistic method, the selection/regeneration of transplastomic plants, and the molecular analyses to select homoplasmic plants and confirm transgene expression.
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Acknowledgments
The authors would like to thank Ms. Lorenza Sannino and Ms. Silvia Silletti for the technical assistance and photography. HIV-1 antisera and recombinant proteins were provided by the Programme EVA Centre for AIDS Reagents, NIBSC, UK, supported by the EC FP6/7 Europrise Network of Excellence, AVIP and NGIN consortia and the Bill and Melinda Gates GHRC-CAVD Project IGV publication no. 368.
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Scotti, N., Cardi, T. (2012). Plastid Transformation as an Expression Tool for Plant-Derived Biopharmaceuticals. In: Dunwell, J., Wetten, A. (eds) Transgenic Plants. Methods in Molecular Biology, vol 847. Humana Press. https://doi.org/10.1007/978-1-61779-558-9_35
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DOI: https://doi.org/10.1007/978-1-61779-558-9_35
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