Pietschmann Thomas
Center for Experimental and Clinical Infection Research, Department of Experimental Virology, Hannover, Germany.
Methods Mol Biol. 2009;510:347-59. doi: 10.1007/978-1-59745-394-3_26.
One hallmark of HCV is its pronounced genetic plasticity, caused by error-prone RNA replication, which probably contributes to its remarkable ability to establish chronic infections. On the basis of phylogenetic analyses, HCV variants are classified into six genotypes (GTs), each comprising a variable number of subtypes. Presumably, these genetic differences, which range from 33 to 35% at the nucleotide level among genotypes and from 22 to 25% between subtypes, are reflected by divergent biological properties of the respective isolates. The unprecedented replication efficiency of the JFH1 isolate (a GT2a strain derived from a Japanese patient with fulminant hepatitis) in transfected Huh-7 cells represents a characteristic feature intrinsic to this particular isolate and has very recently made possible the investigation of the complete viral replication cycle in cultured cells. To expand the scope of this novel HCV infection system, several groups have constructed chimeric HCV genomes comprising JFH1-derived replicase proteins and structural proteins from heterologous HCV strains. This chapter describes experimental procedures for evaluation of the properties of infectious full-length HCV chimeras.
丙型肝炎病毒（HCV）的一个显著特征是其明显的基因可塑性，这是由易出错的RNA复制引起的，这可能有助于其建立慢性感染的显著能力。基于系统发育分析，HCV变体被分为六个基因型（GTs），每个基因型包含数量不等的亚型。据推测，这些基因差异在基因型之间的核苷酸水平上为33%至35%，在亚型之间为22%至25%，各自分离株的不同生物学特性反映了这些差异。JFH1分离株（一种源自日本暴发性肝炎患者的GT2a毒株）在转染的Huh-7细胞中前所未有的复制效率代表了该特定分离株固有的特征，并且最近使得在培养细胞中研究完整的病毒复制周期成为可能。为了扩大这种新型HCV感染系统的范围，几个研究小组构建了嵌合HCV基因组，其包含来自JFH1的复制酶蛋白和来自异源HCV毒株的结构蛋白。本章描述了评估感染性全长HCV嵌合体特性的实验程序。
