Production of single-round infectious chimeric flaviviruses with DNA-based Japanese encephalitis virus replicon
Production of single-round infectious chimeric flaviviruses with DNA-based Japanese encephalitis virus replicon
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DOI：
10.1099/vir.0.058008-0
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发表时间：
2014-01-01
影响因子：
3.8
通讯作者：
Wakita, Takaji
中科院分区：
文献类型：
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作者：
Suzuki, Ryosuke;Ishikawa, Tomohiro;Wakita, Takaji
一种快速制备黄病毒单圆感染性颗粒（srip）的方法将有助于病毒诱变研究。在此，我们建立了一个基于dna的黄病毒条带核酸的生产系统。在巨细胞病毒启动子的控制下，构建了缺乏衣壳前膜包膜C-prM-E编码区的乙型脑炎病毒（JEV）亚基因组复制子质粒。将JEV复制子质粒与JEV C-prM-E表达质粒短暂共转染到293T细胞中，产生了SRIPs，表明与JEV结构蛋白成功反式互补。当C和prM-E蛋白分别提供时，观察到相同的产量水平。此外，登革热1-4型、西尼罗病毒、黄热病或蜱传脑炎病毒prM-E蛋白可用于生产嵌合黄病毒srps，尽管登革热和黄热病病毒的生产效率较低。这些结果表明，我们基于质粒的系统适合于研究黄病毒的生命周期、诊断应用和开发更安全的候选疫苗。
A method for rapid production of single-round infectious particles (SRIPs) of flavivirus would be useful for viral mutagenesis studies. Here, we established a DNA-based production system for SRIPs of flavivirus. We constructed a Japanese encephalitis virus (JEV) subgenomic replicon plasmid, which lacked the C-prM-E (capsid pre-membrane-envelope) coding region, under the control of the cytomegalovirus promoter. When the JEV replicon plasmid was transiently co-transfected with a JEV C-prM-E expression plasmid into 293T cells, SRIPs were produced, indicating successful trans-complementation with JEV structural proteins. Equivalent production levels were observed when C and prM-E proteins were provided separately. Furthermore, dengue types 1-4, West Nile, yellow fever or tick-borne encephalitis virus prM-E proteins could be utilized for production of chimaeric flavivirus SRIPs, although the production was less efficient for dengue and yellow fever viruses. These results indicated that our plasmid-based system is suitable for investigating the life cycles of flaviviruses, diagnostic applications and development of safer vaccine candidates.