Surface display of an internal His-tag on virus-like particles of Nudaurelia capensis omega virus (NomegaV) produced in a baculovirus expression system.
Nudaurelia capensis omega virus (NomegaV) is a member of the Tetraviridae, a family of small, icosahedral, non-enveloped, (+) sense single-stranded RNA insect viruses with T = 4 symmetry. NomegaV virus-like particles (VLPs), which are morphologically indistinguishable from native virions and capable of packaging heterologous RNA, may be produced in the baculovirus expression system. As a first step towards manipulating the tropism of tetraviral nanoparticles (Capsivectors), a (His)6-tag was inserted into the GH loop (between Ala 378 and Gly 379) of the surface-exposed Ig-like domain of NomegaV capsid protein (p70). His-tagged p70 produced in a baculovirus expression system self-assembled into omegaHis VLPs that exhibited similar morphological and RNA encapsidation properties as wild-type NomegaV VLPs produced in the same system. Two assays using paramagnetic pre-charged nickel beads confirmed that multiple affinity tags were present on the surface of omegaHis VLPs and were capable of binding. These results indicate that the GH loop is a suitable site for the retargeting of NomegaV particles for potential biotechnological applications.
Maree
Hans J
HJ
Shimoda Biotech (Pty) Ltd., Greenacres, Port Elizabeth 6057, South Africa. hano@shimoda-biotech.com
van der Walt
Eric
E
Tiedt
Friedrich A C
FA
Hanzlik
Terry N
TN
Appel
Maryke
M
D016367:Baculoviridae / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D005822:Genetic Vectors; D006639:Histidine / Q000235:genetics; D046529:Microscopy, Electron, Transmission; D008958:Models, Molecular; D009842:Oligopeptides / Q000235:genetics; D012328:RNA Viruses / Q000235:genetics; D012367:RNA, Viral / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D014771:Virion / Q000648:ultrastructure*; D019065:Virus Assembly; D014780:Viruses
C471213:His-His-His-His-His-His; D009842:Oligopeptides; D012367:RNA, Viral; D014764:Viral Proteins; D006639:Histidine