Subgenomic replicon derived from a cell line infected with the hepatitis C virus
Subgenomic replicon derived from a cell line infected with the hepatitis C virus
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DOI：
10.1016/s0006-291x(02)00342-x
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发表时间：
2002-05-10
影响因子：
3.1
通讯作者：
Shimotohno, K
中科院分区：
文献类型：
--
作者：
Kishine, H;Sugiyama, K;Shimotohno, K
最近，已经建立了细胞培养系统，其中丙型肝炎病毒（HCV）亚基因组复制子被有效地复制并维持很长一段时间。为了确认来自HCV感染的培养细胞序列的HCV序列是否可用于构建功能性复制子，使用从HCV感染的培养细胞获得的HCV基因组RNA构建了携带新霉素抗性基因的HCV亚基因组RNA。转染后，选择G418抗性Huh-7细胞并亚克隆。最后，在选定的细胞克隆中确认了HCV蛋白的产生和亚基因组RNA的从头合成，表明该亚基因组RNA在培养的细胞中复制并作为复制子起作用。这些结果表明，从体外HCV感染系统与培养的细胞获得的HCV基因组可用于开发具有不同HCV序列的亚基因区复制子系统。(C)2002 Elsevier Science（美国）。All rights reserved.
Recently, cell culture systems have been established, where a hepatitis C virus (HCV) subgenomic replicon was efficiently replicated and maintained for a long period. To see whether a HCV sequence derived from HCV-infected cultured cell sequence can be used for the construction of a functional replicon, a HCV subgenomic RNA carrying a neomycin-resistant gene was constructed using the HCV genome RNA obtained from cultured cells infected with HCV. After transfection, G418-resistant Huh-7 cells were selected and subcloned. Finally, the production of HCV proteins and de novo synthesis of subgenomic RNA were confirmed in the selected cell clone, indicating that this subgenomic RNA replicated in cultured cells and functioned as a replicon. These results suggest that the HCV genome obtained from an in vitro HCV infection system with cultured cells can be used to develop a subgenotnic replicon system with diverse HCV sequences. (C) 2002 Elsevier Science (USA). All rights reserved.