Selected article for: "expression plasmid and protein expression"
Author: Chunyun Sun; Long Chen; Ji Yang; Chunxia Luo; Yanjing Zhang; Jing Li; Jiahui Yang; Jie Zhang; Liangzhi Xie
Title: SARS-CoV-2 and SARS-CoV Spike-RBD Structure and Receptor Binding Comparison and Potential Implications on Neutralizing Antibody and Vaccine Development
  • Document date: 2020_2_20
    • ID: nhq0oq8y_60
    Snippet: Pseudovirus production in 293T adherent cells 6-8 hours before transfection, 293T cells were pre-plate on T75 flask in DMEM+10% FBS at 100,000 cells/cm 2 . 13μg of Luciferase-expressing HIV-1 lentiviral transfer genome (pWPXL-luc), 13μg of packaging plasmid (PSD) and 13μg of expression plasmid encoding either SARS-CoV-2-S protein (pCMV-whCoV-Spike) or SARS-S protein (pCMV-SARS-Spike) were co-transfected into pre-plated 293T cells using Sinofec.....
    Document: Pseudovirus production in 293T adherent cells 6-8 hours before transfection, 293T cells were pre-plate on T75 flask in DMEM+10% FBS at 100,000 cells/cm 2 . 13μg of Luciferase-expressing HIV-1 lentiviral transfer genome (pWPXL-luc), 13μg of packaging plasmid (PSD) and 13μg of expression plasmid encoding either SARS-CoV-2-S protein (pCMV-whCoV-Spike) or SARS-S protein (pCMV-SARS-Spike) were co-transfected into pre-plated 293T cells using Sinofection transfection reagent according to the procedure recommended by manufacturer. Then the transfected cells incubated were incubated at 37℃ and 5% CO 2 overnight, followed by medium exchange with fresh DMEM plus 10% FBS. The supernatant containing pseudovirus was collected after 48-72 hours and filtered through a 0.45μm filter and stored at -80℃ for longtime storage or 4℃ for short time storage.

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