Assembly of bacteriophage Qbeta virus-like particles in yeast Saccharomyces cerevisiae and Pichia pastoris.
Recombinant bacteriophage Qbeta coat protein (CP), which has been proposed as a promising carrier of foreign epitopes via their incorporation either by gene engineering techniques or by chemical coupling, efficiently self-assembles into virus-like particles (VLPs) when expressed in Escherichia coli. Here, we demonstrate expression and self-assembly of Qbeta CP in yeast Saccharomyces cerevisiae and Pichia pastoris. Production reached 3-4 mg/1g of wet cells for S. cerevisiae and 4-6 mg for P. pastoris, which was about 15-20% and 20-30% of the E. coli expression level, respectively. Qbeta VLPs were easily purified by size-exclusion chromatography in both cases and contained nucleic acid, shown by native agarose gel electrophoresis. The obtained particles were highly immunogenic in mice and the resulting sera recognized both E. coli- and yeast-derived Qbeta VLPs equally well.
Freivalds
Janis
J
Biomedical Research and Study Centre, Ratsupites 1, LV-1067 Riga, Latvia.
Dislers
Andris
A
Ose
Velta
V
Skrastina
Dace
D
Cielens
Indulis
I
Pumpens
Paul
P
Sasnauskas
Kestutis
K
Kazaks
Andris
A
D017908:Allolevivirus / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D003001:Cloning, Molecular; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D014771:Virion / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D019065:Virus Assembly
D011994:Recombinant Proteins