The emerging coronavirus disease 2019 (COVID-19) outbreak caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has rapidly spread worldwide resulting in global public health emergencies and economic crises. In this study, a non-infectious and biocontainment level-2 compatible SARS-CoV-2 replicon expressing a nano luciferase (nLuc) reporter was constructed in a bacterial artificial chromosomal (BAC) vector by reverse genetics. The nLuc reporter is highly sensitive, easily quantifiable, and high-throughput adaptable. Upon transfecting the SARS-CoV-2 replicon BAC plasmid DNA into Vero E6 cells, we could detect high levels of nLuc reporter activity and viral RNA transcript, suggesting the replication of the replicon. The replicon replication was further demonstrated by the findings that deleting non-structural protein 15 or mutating its catalytic sites significantly reduced replicon replication, whereas providing the nucleocapsid protein in trans enhanced replicon replication in a dose-dependent manner. Finally, we showed that remdesivir, an FDA-approved antiviral drug, significantly inhibited the replication of the replicon, providing proof-of-principle for the application of our replicon as a useful tool for developing antivirals. Taken together, this study established a sensitive and BSL-2 compatible reporter system in a single BAC plasmid for investigating the functions of SARS-CoV-2 proteins in viral replication and evaluating antiviral compounds. This should contribute to the global effort to combat this deadly viral pathogen. IMPORTANCE The COVID-19 pandemic caused by SARS-CoV-2 is having a catastrophic impact on human lives. Combatting the pandemic requires effective vaccines and antiviral drugs. In this study, we developed a SARS-CoV-2 replicon system with a sensitive and easily quantifiable reporter. Unlike studies involving infectious SARS-CoV-2 virus that must be performed in a biosafety level (BSL) 3 facility, the replicon is non-infectious and thus can be safely used in BSL2 laboratories. The replicon will provide a valuable tool for testing antiviral drugs and studying SARS-CoV-2 biology.

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igh-definition metrology (HDM) systems withflne lateral resolution are capable of capturing the surface shape on a machined part that is beyond the capability of measurement systems employed in manufacturing plants today. Such surface shapes can precisely reflect the impact of cutting processes on surface quality. Understanding the cutting processes and the resultant surface shape is vital to high-precision machining process monitoring and control. This paper presents modeling and experiments of a face milling process to extract surface patterns from measured HDM data and correlate these patterns with cutting force variation. A relationship is established between the instantaneous cutting forces and the observed dominant surface patterns along the feed and circumferential directions for face milling. Potential applications of this relationship in process monitoring, diagnosis, and control are also discussed for face milling. Finally a systematic methodology for characterizing cutting force induced surface variations for a generic machining process is presented by integrating cutting force modeling and HDM measurements.