title: Assembly and Purification of Polyomavirus-Like Particles from Plants.
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.
authors.affiliation: Centre for Biomolecular Engineering, Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, St Lucia, QLD, 4072, Australia.
authors.forename: Emeline V B
authors.initials: EV
authors.lastname: Catrice
authors.forename: Frank
authors.initials: F
authors.lastname: Sainsbury
mesh_terms: D036022:Capsid Proteins / Q000235:genetics* / Q000378:metabolism*; D046529:Microscopy, Electron, Transmission; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000378:metabolism; D011120:Polyomavirus / Q000502:physiology; D017384:Sequence Deletion; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism; D058425:Vaccines, Virus-Like Particle / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D019065:Virus Assembly
chemical_list: D036022:Capsid Proteins; C062481:VP1 protein, polyomavirus; D058425:Vaccines, Virus-Like Particle