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RNA-Directed Assembly of Tobacco Mosaic Virus (TMV)-Like Carriers with Tunable Fractions of Differently Addressable Coat Proteins

Part of the Methods in Molecular Biology book series (MIMB,volume 1776)

Abstract

Taking advantage of the ability for in vitro assembly of the plant-infecting virus tobacco mosaic virus (TMV), rod-shaped nanoscale scaffolds presenting different addressable groups can be obtained. We have established procedures resulting in virus-like particles with randomly distributed functional groups, with different groups arranged in striped but randomized structures, and even with distinct groups clustered in adjacent, better-defined domains. The TMV coat protein (CP) variants combined in these approaches can either originate all from TMV mutants propagated in planta, or be mixed with CP expressed in E. coli (CPEc). Protocols for expression and purification of a CPEc-His6 mutant in E. coli as well as the different methods for in vitro assembly and the visualization by decoration of one CP type are explained in detail.

Key words

  • Tobacco mosaic virus (TMV)
  • Multifunctional
  • In vitro assembly
  • TMV decoration
  • Transmission electron microscopy

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Acknowledgments

I would like to thank Fania Geiger and Fabian Eber for establishing the protocols for sequential assembly and decoration of TMV, respectively. I am also grateful for the work of Diether Gotthardt, our gardener, and Sigrid Kober for taking care of the plants and virus isolations. Special thanks to Holger Jeske and Christina Wege for their great support and without whom there would be no plant virus nanotechnology in Stuttgart. This work was financed in part by the DFG PAK 410 and SPP1569, the Zeiss foundation, “Projekthaus” NanoBioMater as well as the Baden Württemberg Stiftung in the course of the Network of Competence “Functional Nanostructures.”

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Eiben, S. (2018). RNA-Directed Assembly of Tobacco Mosaic Virus (TMV)-Like Carriers with Tunable Fractions of Differently Addressable Coat Proteins. In: Wege, C., Lomonossoff, G. (eds) Virus-Derived Nanoparticles for Advanced Technologies. Methods in Molecular Biology, vol 1776. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7808-3_3

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  • DOI: https://doi.org/10.1007/978-1-4939-7808-3_3

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