Novel application of neferine to inhibition of SARS-CoV and SARS-CoV-2 infection
A technology of lysinine and sars-cov-2, applied in the field of natural medicine, can solve the problems of many potential hosts and long incubation period, etc.
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[0024] 1) Construct SARS-CoV, SARS-CoV-2, VSV-G pseudovirus system
[0025] The SARS-CoV, SARS-CoV-2 and VSV-G pseudovirus systems were constructed using the three-plasmid packaging system of lentivirus. The pseudovirus system uses SARS-CoV or SARS-CoV-2 spike protein or VSV G protein as surface membrane protein, and firefly luciferase as reporter gene. Using GenJet Ver.II transfection reagent, spike envelope plasmid or G protein envelope plasmid, packaging plasmid psPAX2, and HIV plasmid containing luciferase reporter gene were co-transfected into HEK293T cells, and fresh culture medium was replaced after 12 hours. After 24, 36, 48, and 60 hours of transfection, collect the supernatant culture solution, centrifuge it at 800×g for 5 minutes, and then filter it through a 0.45 μm filter membrane to remove cell debris. The obtained pseudovirus is subpackaged and stored in - 80°C.
[0026] 2) The whole process of administration of nexinine to study its inhibitory effect on HEK29...
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