Genotype 1b strain Con1 represents an important reference in the study of hepatitis C virus (HCV). Here, we aimed to develop an advanced infectious Con1 recombinant. We found that previously identified mutations A1226G/F1464L/A1672S/Q1773H permitted culture adaption of Con1 Core-NS5A (C-5A) recombinant containing 5’UTR and NS5B-3’UTR from JFH1 (genotype 2a), thus acquired additional mutations L725H/F886L/D2415G. C-5A containing all seven mutations (C-5A_7m) replicated efficiently in Huh7.5 and Huh7.5.1 cells and had an increased infectivity in SEC14L2-expressing Huh7.5.1 cells. Incorporation of Con1 NS5B was deleterious to C-5A_7m, however Con1 5’UTR was permissive but attenuated the virus. Nucleotides G1, A4, and G35 primarily accounted for the viral attenuation without affecting RNA translation. C-5A_7m was inhibited dose-dependently by simeprevir and daclatasvir, and substitutions at A4, A29, A34, and G35 conferred resistance to miR-122 antagonism. The novel Con1 5’UTR-NS5A recombinant, adaptive mutations, and critical nucleotides described here will facilitate future studies of HCV culture systems and virus-host interaction.

基因型 1b 菌株 Con1 代表了丙型肝炎病毒 (HCV) 研究的重要参考。在这里，我们的目标是开发一种先进的传染性 Con1 重组体。我们发现先前鉴定的突变 A1226G/F1464L/A1672S/Q1773H 允许来自 JFH1（基因型 2a）的含有 5'UTR 和 NS5B-3'UTR 的 Con1 Core-NS5A (C-5A) 重组体的培养适应，因此获得了额外的突变 L725H/ F886L/D2415G。包含所有七种突变的 C-5A (C-5A_7m) 在 Huh7.5 和 Huh7.5.1 细胞中有效复制，并且在表达 SEC14L2 的 Huh7.5.1 细胞中具有增加的感染性。Con1 NS5B 的掺入对 C-5A_7m 有害，但 Con1 5'UTR 是允许的，但会减弱病毒。核苷酸 G1、A4 和 G35 主要解释了病毒减毒而不影响 RNA 翻译。simeprevir 和 daclatasvir 对 C-5A_7m 的抑制呈剂量依赖性，并且 A4、A29、A34 和 G35 的取代赋予了对 miR-122 拮抗剂的抗性。此处描述的新型 Con1 5'UTR-NS5A 重组、适应性突变和关键核苷酸将有助于未来对 HCV 培养系统和病毒-宿主相互作用的研究。