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. 2020 Oct 15;11:5214. doi: 10.1038/s41467-020-19055-7

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© The Author(s) 2020, corrected publication 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — a Assembly of the full-length SARS-CoV-2-Nluc cDNA. The Nanoluciferase (Nluc) gene together with a PacI site was placed downstream of the regulatory sequence of ORF7 to replace the ORF7 sequence. The nucleotide identities of the Nluc substitution sites are indicated. b Plaque morphologies of infectious clone-derived P1 SARS-CoV-2-Nluc (P1 IC Nluc) and wild-type SARS-CoV-2 (IC WT). c Replication kinetics. Vero E6 cells were infected with infectious clone-derived IC WT or P1 IC Nluc at MOI 0.01. Viruses in culture supernatants were quantified by plaque assay. The means ± standard deviations from three independent experiments are shown. Two-way ANOVA with correct for multiple comparisons are used for statistical analyses. d Plaque morphology of P5 IC Nluc. e Replication kinetics of P5 IC Nluc on Vero E6 cells. The means ± standard deviations from three independent experiments are shown. Two-way ANOVA with correct for multiple comparisons are used for statistical analyses. f Luciferase signals produced from SARS-CoV-2-Nluc-infected Vero E6 cells at 12 h post-infection. Cells were infected with viruses at MOI 0.1. The means ± standard deviations from six independent experiments are shown. One-way ordinary ANOVA test was used for statistical analyses. g Gel analysis of IC Nluc virus stability. The left panel depicts the theoretical results of RT-PCR followed by restriction enzyme digestion. The right panel shows the gel analysis of the RT-PCR products before (lanes 1–3) and after BsrGI/PacI digestion (lanes 4–6). h Summary of full-genome sequences of P1 and P5 IC Nluc viruses. Nucleotide and amino acid differences from the IC WT are indicated.