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目的:构建人β冠状病毒程序性核糖体移码效率评价系统，为筛选靶向核糖体移码的抗病毒宿主因子及药物提供工具。方法:比对分析冠状病毒开放阅读框（open reading frame，ORF）1a和1b之间的核糖体移码序列，构建真核细胞表达双荧光报告质粒，通过共转染报告质粒与宿主因子表达质粒，验证宿主因子对人β冠状病毒程序性核糖体移码的影响。结果:成功构建人β冠状病毒程序性核糖体移码真核细胞表达双荧光报告检测系统，通过该系统鉴定干扰素刺激基因产物C19orf66（称为"Shiftless"）是抑制新型冠状病毒（SARS-CoV-2）程序性核糖体移码的宿主因子。结论:感染人的5种β冠状病毒中程序性核糖体移码位点进化保守，本研究中程序性核糖体移码效率评价系统的构建策略可广泛应用于抗冠状病毒药物筛选及病毒宿主互作研究。

Objective:To construct an efficiency evaluation system for the programmed ribosomal frameshift of human β-coronaviruses, so as to provide a tool for screening antiviral host factors and drugs targeting ribosomal frameshift.Methods:The ribosomal frameshift sequences between coronavirus open reading frame (ORF) 1a and 1b were analyzed. The eukaryotic expression dual fluorescent reporter plasmids were constructed, and the influence of host factors on the programmed ribosome frameshift of human β-coronaviruses was verified by co-transfection of the reporter plasmid and the host factor expression plasmid.Results:A eukaryotic cell expressed double fluorescence reporting and detection system for programmed ribosomal frameshift of human β-coronaviruses were constructed successfully. The interferon-stimulated gene product C19orf66 (also called Shiftless) was identified as a host factor inhibiting the programmed ribosome frameshift of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).Conclusions:Programmed ribosome frameshift slippery site in five human β-coronaviruses are evolutionally-conserved. The strategy for constructing the frameshift efficiency evaluation system in this study can be widely used in anti-coronavirus drug screening and virus-host interaction studies.