title: Expression in tobacco and purification of beak and feather disease virus capsid protein fused to elastin-like polypeptides.
abstract: Psittacine beak and feather disease, caused by beak and feather disease virus (BFDV), is a threat to endangered psittacine species. There is currently no vaccine against BFDV, which necessitates the development of safe and affordable vaccine candidates. A subunit vaccine based on BFDV capsid protein (CP), the major antigenic determinant, expressed in the inexpensive and highly scalable plant expression system could satisfy these requirements. Full-length CP and a truncated CP (ΔN40 CP) were transiently expressed in tobacco (Nicotiana benthamiana) as fusions to elastin-like polypeptide (ELP). These two proteins were fused to ELPs of different lengths in order to increase expression levels and to provide a simple means of purification. The ELP fusion proteins were purified by inverse transition cycling (ITC) and it was found that a membrane filtration-based ITC method improved the recovery of ΔN40 CP-ELP51 fusion protein relative to a centrifugation-based method.
authors.affiliation: Department of Molecular and Cell Biology, University of Cape Town, Rondebosch 7700, South Africa.
authors.forename: Lucian
authors.initials: L
authors.lastname: Duvenage
authors.forename: Inga I
authors.initials: II
authors.lastname: Hitzeroth
authors.forename: Ann E
authors.initials: AE
authors.lastname: Meyers
authors.forename: Edward P
authors.initials: EP
authors.lastname: Rybicki
mesh_terms: D000818:Animals; D001709:Biotechnology / Q000379:methods; D036022:Capsid Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D017925:Circovirus / Q000235:genetics*; D004549:Elastin / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D005374:Filtration / Q000379:methods; D015870:Gene Expression; D008967:Molecular Biology / Q000379:methods; D030821:Plants, Genetically Modified; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D014026:Nicotiana / Q000235:genetics*; D014765:Viral Vaccines / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
chemical_list: D036022:Capsid Proteins; D011993:Recombinant Fusion Proteins; D014765:Viral Vaccines; D004549:Elastin