Grun J B, Brinton M A
Wistar Institute, Philadelphia, Pennsylvania 19104.
J Gen Virol. 1988 Dec;69 ( Pt 12):3121-7. doi: 10.1099/0022-1317-69-12-3121.
Flaviviruses encode seven non-structural proteins for which functions have not yet been described. The identification of the viral and possible host proteins which may be involved in flavivirus replication has been impeded by the fact that the viral replication complexes are tightly associated with endoplasmic reticular membranes within infected cells and that in vitro polymerase activity is associated with large membrane fragments. To facilitate further study of flavivirus replication complexes, selected ultrapure detergents were analysed for their effect on West Nile virus (WNV) in vitro RNA-dependent RNA polymerase activity and for their ability to release functional replication complexes from partially purified intracellular BHK-21 membrane fragments. A few previous reports indicated that flavivirus in vitro polymerase activity was sensitive to detergent treatment. The present study indicates that WNV polymerase activity is variably inhibited depending on the concentration and identity of the detergent used. Of the five detergents (Tween 20, maltoside, octylglucoside, lubrol PX and sodium deoxycholate) tested, sodium deoxycholate was the most efficient at releasing functional viral replication complexes from intracellular membranes.
黄病毒编码七种非结构蛋白，其功能尚未明确。病毒复制复合体与受感染细胞内的内质网膜紧密相连，且体外聚合酶活性与大的膜片段相关，这一事实阻碍了对可能参与黄病毒复制的病毒及潜在宿主蛋白的鉴定。为便于进一步研究黄病毒复制复合体，分析了几种选定的超纯去污剂对西尼罗河病毒（WNV）体外RNA依赖性RNA聚合酶活性的影响，以及它们从部分纯化的细胞内BHK-21膜片段中释放功能性复制复合体的能力。之前有一些报道表明，黄病毒体外聚合酶活性对去污剂处理敏感。本研究表明，WNV聚合酶活性受到的抑制程度因所用去污剂的浓度和种类而异。在所测试的五种去污剂（吐温20、麦芽糖苷、辛基葡糖苷、月桂醇聚醚-10和脱氧胆酸钠）中，脱氧胆酸钠最有效地从细胞内膜中释放出功能性病毒复制复合体。
