Establishment of infectious HCV virion-producing cells with newly designed full-genome replicon RNA.
Establishment of infectious HCV virion-producing cells with newly designed full-genome replicon RNA.
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DOI：
10.1007/s00705-010-0859-x
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发表时间：
2011-02
影响因子：
2.7
通讯作者：
Kohara, Michinori
中科院分区：
文献类型：
--
作者：
Arai, Masaaki;Suzuki, Hidenori;Tobita, Yoshimi;Takagi, Asako;Okamoto, Koichi;Ohta, Atsunori;Sudoh, Masayuki;Kohara, Michinori
丙型肝炎病毒 (HCV) 复制子系统能够深入分析 HCV 的生命周期。然而，之前报道的全基因组复制子系统无法产生真正的病毒体。基于这些结果，我们构建了新设计的全基因组复制子RNA，它由延伸至NS2区域的完整5'端半RNA组成，两侧是额外的选择标记基因。含有这种全基因组 RNA 的 Huh-7 细胞在 G418 选择下增殖良好，并将病毒粒子样颗粒分泌到上清液中。通过电子显微镜分析，这些颗粒呈圆形，直径为 50 nm，浮力密度为 1.08 g/mL，经 NP-40 处理后变为 1.19 g/mL；这些数字与完整病毒体和无包膜核衣壳的推定密度相匹配。这些颗粒还在菌落形成试验中显示出感染性。该系统可能为研究丙型肝炎病毒的生命周期提供另一种选择。
Hepatitis C virus (HCV) replicon systems enable in-depth analysis of the life cycle of HCV. However, the previously reported full-genome replicon system is unable to produce authentic virions. On the basis of these results, we constructed newly designed full-genomic replicon RNA, which is composed of the intact 5′-terminal-half RNA extending to the NS2 region flanked by an extra selection marker gene. Huh-7 cells harboring this full-genomic RNA proliferated well under G418 selection and secreted virion-like particles into the supernatant. These particles, which were round and 50 nm in diameter when analyzed by electron microscopy, had a buoyant density of 1.08 g/mL that shifted to 1.19 g/mL after NP-40 treatment; these figures match the putative densities of intact virions and nucleocapsids without envelope. The particles also showed infectivity in a colony-forming assay. This system may offer another option for investigating the life cycle of HCV.
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DOI：
10.1073/pnas.0504877103
发表时间：
2006-05-09
影响因子：
11.1
作者：
Pietschmann, Thomas;Kaul, Artur;Bartenschlager, Ralf
通讯作者：
Bartenschlager, Ralf
影响因子：
5.4
作者：
Ikeda, M;Yi, MK;Lemon, SA
通讯作者：
Lemon, SA
影响因子：
5.4
作者：
Kanda, T;Basu, A;Ray, RB
通讯作者：
Ray, RB
影响因子：
5.4
作者：
Blight, KJ;McKeating, JA;Rice, CM
通讯作者：
Rice, CM
影响因子：
56.9
作者：
Pileri, P;Uematsu, Y;Abrignani, S
通讯作者：
Abrignani, S