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.
关键词
冠状病毒/程序性核糖体移码/RNA假结/宿主因子/双荧光报告系统
Key words
Coronavirus/Programmed ribosome frameshift/RNA pseudoknot/Host factor/Dual fluorescence reporting system
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