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

• Published in: Journal of virological methods Vol. 136; no. 1; pp. 283 - 288
• Main Authors: Maree, Hans J., van der Walt, Eric, Tiedt, Friedrich A.C., Hanzlik, Terry N., Appel, Maryke
• Format: Journal Article
• Language: English
• Published: London Elsevier B.V 01.09.2006; Amsterdam Elsevier; New York, NY
• Subjects: Baculoviridae - genetics; Baculovirus; Baculovirus expression; Biological and medical sciences; Electrophoresis, Polyacrylamide Gel; Fundamental and applied biological sciences. Psychology; Genetic Vectors; His-tag; Histidine - genetics; Microbiology; Microscopy, Electron, Transmission; Models, Molecular; Nudaurelia capensis ^w virus; Nudaurelia capensis ω virus; Oligopeptides - genetics; RNA Viruses - genetics; RNA, Viral - metabolism; Surface display; Techniques used in virology; Tetraviridae; Tetravirus; Viral Proteins - genetics; Viral Proteins - metabolism; Virion - ultrastructure; Virology; Virus Assembly; Virus-like particle; Viruses; Baculovirus expression; Virus-like particle; Nudaurelia capensis ω virus; Tetravirus; Surface display; His-tag; Virus; Baculoviridae; Microbiology; Virus like particle; Nudaurelia capensis to virus; Method; Virology
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2006.05.014

Nudaurelia capensis ω virus (NωV) is a member of the Tetraviridae, a family of small, icosahedral, non-enveloped, (+) sense single-stranded RNA insect viruses with T = 4 symmetry. NωV 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 NωV capsid protein (p70). His-tagged p70 produced in a baculovirus expression system self-assembled into ωHis VLPs that exhibited similar morphological and RNA encapsidation properties as wild-type NωV 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 ωHis VLPs and were capable of binding. These results indicate that the GH loop is a suitable site for the retargeting of NωV particles for potential biotechnological applications.