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Isolation and Characterization of Two Distinct Types of Unmodified Spherical Plant Sobemovirus-Like Particles for Diagnostic and Technical Uses

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

Abstract

Plant virus-like particles (VLPs) structurally resemble their progenitor viruses, but are noninfectious due to absence of viral nucleic acids. Since the 1980s, VLPs have been actively studied with the aim of constructing different nanomaterials, including immunologically active carriers for peptides and whole proteins and proteinaceous shells for the packaging of different ligands.

The technological developments using VLPs require large amounts of purified particles. Here, we describe the laboratory process for isolation and purification of two unmodified plant VLPs, derived from two sobemoviruses, cocksfoot mottle virus (CfMV) and rice yellow mottle virus (RYMV), which is based on cultivation of recombinant Escherichia coli cells, VLP precipitation from bacterial extracts and ultracentrifugation. The suggested purification scheme allows the production of 4–45 mg of purified sobemoviral VLPs from a 1 l bacterial culture, depending on the required purity level. Additionally, we provide short protocols for VLP characterization using SDS-PAGE, agarose gel electrophoresis, ultraviolet and mass spectrometry, dynamic light scattering, and electron microscopy.

Key words

  • Virus-like
  • Sobemovirus
  • Cocksfoot mottle virus
  • Rice yellow mottle virus
  • Ultracentrifugation

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Acknowledgment

We wish to thank Dr. V. Ose for help in the preparation of EM pictures and J. Bogans for DLS experiments. G. Grinberga and V. Zeltina are acknowledged for their technical assistance. The chapter was written with the help of Grant No. SP 672/2014 provided by the Latvian Science Council.

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Correspondence to Andris Zeltins .

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Balke, I., Reseviča, G., Zeltins, A. (2018). Isolation and Characterization of Two Distinct Types of Unmodified Spherical Plant Sobemovirus-Like Particles for Diagnostic and Technical Uses. 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_2

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  • DOI: https://doi.org/10.1007/978-1-4939-7808-3_2

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7806-9

  • Online ISBN: 978-1-4939-7808-3

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