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Author:
Date:
2014-05-20
[Abstract] As for all positive strand RNA viruses, hepatitis C virus (HCV) RNA replication is tightly associated with rearranged host cell membranes, termed viral replication factories. However, up to now little is known about both viral and cellular constituents of viral replication factories. Here, we describe a protocol to specifically isolate HCV-remodeled host cell membranes and endoplasmic reticulum (ER) membranes of naïve cells, by using a functional NS4B HA-tagged subgenomic replicon and a C-terminally HA-tagged calnexin-overexpressing cell line, respectively. Post-nuclear whole cell membrane ...
[摘要] 至于所有正链RNA病毒，丙型肝炎病毒（HCV）RNA复制与称为病毒复制工厂的重排宿主细胞膜紧密相关。 然而，到目前为止对病毒复制工厂的病毒和细胞成分了解甚少。 在这里，我们描述一个协议，通过使用功能NS4B HA标记亚基因组复制子和C末端HA标记calnexin过表达细胞系，特异性隔离HCV重塑的宿主细胞膜和内质网（ER）膜的幼稚细胞， 分别。 首先通过密度梯度离心富集核后全细胞膜级分，随后通过HA特异性亲和标签纯化。 在天然条件下洗脱时，纯化的样品可进行多种生物化学和功能测定。
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