Maree Hans J, van der Walt Eric, Tiedt Friedrich A C, Hanzlik Terry N, Appel Maryke
Shimoda Biotech (Pty) Ltd., Greenacres, Port Elizabeth 6057, South Africa.
J Virol Methods. 2006 Sep;136(1-2):283-8. doi: 10.1016/j.jviromet.2006.05.014. Epub 2006 Jun 23.
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.
卡普斯角无尾大蚕蛾ω病毒(NomegaV)是四病毒科的成员,该科是一类小型、二十面体、无包膜、正链单链RNA昆虫病毒,具有T = 4对称性。NomegaV病毒样颗粒(VLP)在形态上与天然病毒粒子无法区分,并且能够包装异源RNA,可以在杆状病毒表达系统中产生。作为操纵四病毒纳米颗粒(Capsivectors)嗜性的第一步,在NomegaV衣壳蛋白(p70)表面暴露的Ig样结构域的GH环(在Ala 378和Gly 379之间)中插入了一个(His)6标签。在杆状病毒表达系统中产生的His标签化p70自组装成omegaHis VLP,其表现出与在同一系统中产生的野生型NomegaV VLP相似的形态和RNA包装特性。使用顺磁预充电镍珠的两种测定方法证实,omegaHis VLP表面存在多个亲和标签并且能够结合。这些结果表明,GH环是用于将NomegaV颗粒重新靶向潜在生物技术应用的合适位点。