French T J, Marshall J J, Roy P
NERC Institute of Virology & Environmental Microbiology, Oxford, United Kingdom.
J Virol. 1990 Dec;64(12):5695-700. doi: 10.1128/JVI.64.12.5695-5700.1990.
Bluetongue is a disease of ruminants. The etiologic agent is bluetongue virus (BTV), a gnat-transmitted member of the Orbivirus genus of the Reoviridae. The virus has a genome of 10 double-stranded RNA species L1 to L3, M4 to M6, S7 to S10). The L2 and M5 genes of BTV which encode the outer capsid proteins VP2 and VP5, respectively, were inserted into a recombinant baculovirus downstream of duplicated copies of the baculovirus polyhedrin promoter. Insect cells coinfected with this virus plus a recombinant baculovirus expressing the two major core proteins VP3 and VP7 of BTV (T.J. French and P. Roy, J. Virol. 64:1530-1536, 1990) synthesized noninfectious, double-shelled, viruslike particles. When purified, these particles were found to have the same size and appearance as authentic BTV virions and exhibited high levels of hemagglutination activity. Antibodies raised to the expressed particles contained high titers of neutralizing activity against the homologous BTV serotype. The assembly of these bluetongue viruslike particles after the simultaneous expression of four separate proteins is indicative of the potential of this technology for the production of a new generation of viral vaccines and for the study of complex, multiprotein structures.
蓝舌病是反刍动物的一种疾病。病原体是蓝舌病病毒（BTV），它是呼肠孤病毒科环状病毒属中由蚋传播的一种病毒。该病毒的基因组由10种双链RNA组成（L1至L3、M4至M6、S7至S10）。将分别编码外衣壳蛋白VP2和VP5的BTV的L2和M5基因插入杆状病毒多角体蛋白启动子重复拷贝下游的重组杆状病毒中。用这种病毒与表达BTV两种主要核心蛋白VP3和VP7的重组杆状病毒共同感染昆虫细胞（T.J. French和P. Roy，《病毒学杂志》64:1530 - 1536，1990年），合成了无感染性的、双层的、病毒样颗粒。纯化后发现这些颗粒的大小和外观与真正的BTV病毒粒子相同，并表现出高水平的血凝活性。针对表达颗粒产生的抗体含有高滴度的针对同源BTV血清型的中和活性。在同时表达四种不同蛋白质后组装这些蓝舌病病毒样颗粒，表明该技术在生产新一代病毒疫苗以及研究复杂的多蛋白结构方面具有潜力。
