Yoshii Kentarou, Hayasaka Daisuke, Goto Akiko, Kawakami Kazue, Kariwa Hiroaki, Takashima Ikuo
Laboratory of Public Health, Department of Environmental Veterinary Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Kita-18 Nishi-9, Kita-ku, Sapporo, Hokkaido 060-0818, Japan.
Vaccine. 2005 Jun 10;23(30):3946-56. doi: 10.1016/j.vaccine.2005.03.004. Epub 2005 Mar 25.
The sub-genomic replicon of tick-borne encephalitis (TBE) virus (Far-Eastern subtype) was packaged into infectious particles by providing the viral structural proteins in trans. Sequential transfection of TBE replicon RNA and a plasmid that expressed the structural proteins led to the secretion of infectious particles that contained TBE replicon RNA. The secreted particles had single-round infectivity, which was inhibited by TBE virus-neutralizing antibody. The physical structure of the particles was almost identical to that of infectious virions, and the packaged replicon RNA showed no recombination with the mRNAs of the viral structural proteins. Furthermore, heterologous genes were successfully delivered and expressed by packaging TBE replicon RNA with inserted GFP and Neo genes. This replicon packaging system may be a useful tool for the molecular study of the TBE virus genome packaging mechanism, and for the development of vaccine delivery systems.
通过反式提供病毒结构蛋白，将蜱传脑炎（TBE）病毒（远东亚型）的亚基因组复制子包装成感染性颗粒。将TBE复制子RNA和表达结构蛋白的质粒进行顺序转染，导致分泌出含有TBE复制子RNA的感染性颗粒。分泌的颗粒具有单轮感染性，可被TBE病毒中和抗体抑制。颗粒的物理结构与感染性病毒粒子几乎相同，并且包装的复制子RNA与病毒结构蛋白的mRNA没有重组。此外，通过包装插入了绿色荧光蛋白（GFP）和新霉素（Neo）基因的TBE复制子RNA，成功递送并表达了异源基因。这种复制子包装系统可能是研究TBE病毒基因组包装机制的分子研究以及开发疫苗递送系统的有用工具。
