title: Synthesis of empty african horse sickness virus particles.
abstract: As a means to develop African horse sickness (AHS) vaccines that are safe and DIVA compliant, we investigated the synthesis of empty African horse sickness virus (AHSV) particles. The emphasis of this study was on the assembly of the major viral core (VP3 and VP7) and outer capsid proteins (VP2 and VP5) into architecturally complex, heteromultimeric nanosized particles. The production of fully assembled core-like particles (CLPs) was accomplished in vivo by baculovirus-mediated co-synthesis of VP3 and VP7. The two different outer capsid proteins were capable of associating independently of each other with preformed cores to yield partial virus-like particles (VLPs). Complete VLPs were synthesized, albeit with a low yield. Crystalline formation of AHSV VP7 trimers is thought to impede high-level CLP production. Consequently, we engineered and co-synthesized VP3 with a more hydrophilic mutant VP7, resulting in an increase in the turnover of CLPs.
authors.affiliation: Transboundary Animal Diseases, and Molecular Epidemiology and Diagnostics, Onderstepoort Veterinary Institute, Agricultural Research Council, Pretoria 0110, South Africa. Electronic address: marees@arc.agric.za.
authors.forename: Sonja
authors.initials: S
authors.lastname: Maree
authors.affiliation: Transboundary Animal Diseases, Onderstepoort Veterinary Institute, Agricultural Research Council, Pretoria 0110, South Africa; Department of Microbiology and Plant Pathology, University of Pretoria, Pretoria 0002, South Africa.
authors.forename: Francois F
authors.initials: FF
authors.lastname: Maree
authors.affiliation: Electron Microscopy Unit, Onderstepoort Veterinary Institute, Agricultural Research Council, Pretoria 0110, South Africa.
authors.forename: John F
authors.initials: JF
authors.lastname: Putterill
authors.affiliation: European Molecular Biology Laboratory, European Bioinformatics Institute (EMBL-EBI), Wellcome Trust Genome Campus, Hinxton, Cambridge CB10 1SD, United Kingdom; Biozentrum, University of Basel, Basel 4056, Switzerland; Swiss Institute of Bioinformatics (SIB), Basel 4056, Switzerland.
authors.forename: Tjaart A P
authors.initials: TAP
authors.lastname: de Beer
authors.affiliation: Department of Genetics, University of Pretoria, Pretoria 0002, South Africa.
authors.forename: Henk
authors.initials: H
authors.lastname: Huismans
authors.affiliation: Department of Microbiology and Plant Pathology, University of Pretoria, Pretoria 0002, South Africa.
authors.forename: Jacques
authors.initials: J
authors.lastname: Theron
mesh_terms: D000356:African Horse Sickness Virus / Q000235:genetics*; D016367:Baculoviridae; D005822:Genetic Vectors; D055503:Protein Multimerization; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D058425:Vaccines, Virus-Like Particle / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D015678:Viral Structural Proteins / Q000235:genetics / Q000378:metabolism; D022701:Virosomes / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D058425:Vaccines, Virus-Like Particle; D015678:Viral Structural Proteins; D022701:Virosomes
keywords: AHSV; African horse sickness virus; Core-like particles; VLP; Virus-like particles