Assembly and purification of polyomavirus-like particles from plants

Catrice, Emeline V. B. and Sainsbury, Frank (2015) Assembly and purification of polyomavirus-like particles from plants. Molecular Biotechnology, 57 10: 904-913. doi:10.1007/s12033-015-9879-9


Author Catrice, Emeline V. B.
Sainsbury, Frank
Title Assembly and purification of polyomavirus-like particles from plants
Journal name Molecular Biotechnology   Check publisher's open access policy
ISSN 1559-0305
1073-6085
Publication date 2015-07-16
Sub-type Article (original research)
DOI 10.1007/s12033-015-9879-9
Volume 57
Issue 10
Start page 904
End page 913
Total pages 10
Place of publication New York, United States
Publisher Humana Press
Collection year 2016
Language eng
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.
Keyword Virus-like particle
Recombinant protein
Molecular farming
Transient expression
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Published online 16 July 2015.

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2016 Collection
Australian Institute for Bioengineering and Nanotechnology Publications
 
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