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丙型肝炎病毒复制子:概述与基本实验方案

HCV replicons: overview and basic protocols.

作者信息

Lohmann Volker

机构信息

University of Heidelberg, Heidelberg, Germany.

出版信息

Methods Mol Biol. 2009;510:145-63. doi: 10.1007/978-1-59745-394-3_11.

Abstract

Subgenomic replicons have been the first efficient cell-culture system for HCV and still are a valuable tool for studying different aspects of RNA replication. A variety of replicons based on different viral isolates and vector designs have been established. Here, I give a brief overview of viral isolates, applicable host-cell lines, replicon structures, and general considerations regarding replicon experiments, supplemented by basic protocols for in vitro transcription, electroporation, selection of replicon cells, transient replication assays, and northern hybridization.

摘要

亚基因组复制子一直是丙型肝炎病毒(HCV)首个有效的细胞培养系统,至今仍是研究RNA复制各个方面的宝贵工具。基于不同病毒分离株和载体设计的多种复制子已被构建。在此,我简要概述病毒分离株、适用的宿主细胞系、复制子结构以及复制子实验的一般注意事项,并辅以体外转录、电穿孔、复制子细胞筛选、瞬时复制测定和Northern杂交的基本方案。

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