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Assembly and Purification of Polyomavirus-Like Particles from Plants

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Abstract

Polyomaviruses are small DNA viruses that have a history of use in biotechnology. The capsids of a number of species have been developed into experimental prophylactic and therapeutic virus-like particle (VLP) vaccines. In order to explore plants as a host for the expression and purification of polyomavirus-like particles, we have transiently expressed the major capsid protein, VP1, in Nicotiana benthamiana leaves. Deletion of a polybasic motif from the N-terminal region of VP1 resulted in increased expression as well as reduced necrosis of leaf tissue, which was associated with differences in subcellular localisation and reduced DNA binding by the deletion variant (ΔVP1). Self-assembled VLPs were recovered from tissue expressing both wild-type VP1 and ΔVP1 by density gradient ultracentrifugation. VLPs composed of ΔVP1 were more homogenous than wtVPLs and, unlike the latter, did not encapsidate nucleic acid. Such homogenous, empty VLPs are of great interest in biotechnology and nanotechnology. In addition, we show that both MPyV VLP variants assembled in plants can be produced with encapsidated foreign protein. Thus, this study demonstrates the utility of plant-based expression of polyomavirus-like particles and the suitability of this host for further developments in polyomavirus-based technologies.

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Abbreviations

AF4:

Asymmetric flow field-flow fractionation

DLS:

Dynamic light scattering

Dpi:

Days post-infiltration

GFP:

Green fluorescent protein

NLS:

Nuclear localisation signal

PBS:

Phosphate-buffered saline

TEM:

Transmission electron microscopy

VLP:

Virus-like particle

wt:

Wild-type

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Acknowledgments

The authors would like to thank Dr. Alexandra Depelsenaire for assistance with confocal microscopy. Professor George Lomonossoff and Plant Biosciences Limited are thanked for providing the pEAQ-HT plasmid.

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Authors and Affiliations

  1. Centre for Biomolecular Engineering, Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, St Lucia, QLD, 4072, Australia

    Emeline V. B. Catrice & Frank Sainsbury

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  1. Emeline V. B. Catrice
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  2. Frank Sainsbury
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Correspondence to Frank Sainsbury.

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Catrice, E.V.B., Sainsbury, F. Assembly and Purification of Polyomavirus-Like Particles from Plants. Mol Biotechnol 57, 904–913 (2015). https://doi.org/10.1007/s12033-015-9879-9

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