Surface display of an internal His-tag on virus-like particles of Nudaurelia capensis omega virus (NomegaV) produced in a baculovirus expression system

J Virol Methods. 2006 Sep;136(1-2):283-8. doi: 10.1016/j.jviromet.2006.05.014. Epub 2006 Jun 23.

Abstract

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.

MeSH terms

  • Baculoviridae / genetics*
  • Electrophoresis, Polyacrylamide Gel
  • Genetic Vectors*
  • Histidine / genetics
  • Microscopy, Electron, Transmission
  • Models, Molecular
  • Oligopeptides / genetics
  • RNA Viruses* / genetics
  • RNA, Viral / metabolism
  • Viral Proteins / genetics
  • Viral Proteins / metabolism*
  • Virion / ultrastructure*
  • Virus Assembly
  • Viruses

Substances

  • His-His-His-His-His-His
  • Oligopeptides
  • RNA, Viral
  • Viral Proteins
  • Histidine