Lohmann Volker
University of Heidelberg, Heidelberg, Germany.
Methods Mol Biol. 2009;510:145-63. doi: 10.1007/978-1-59745-394-3_11.
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.
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。
