Selected article for: "absence presence and luciferase reporter"
Author: Xia, Shuai; Yan, Lei; Xu, Wei; Agrawal, Anurodh Shankar; Algaissi, Abdullah; Tseng, Chien-Te K.; Wang, Qian; Du, Lanying; Tan, Wenjie; Wilson, Ian A.; Jiang, Shibo; Yang, Bei; Lu, Lu
Title: A pan-coronavirus fusion inhibitor targeting the HR1 domain of human coronavirus spike
  • Document date: 2019_4_10
    • ID: 3c5ab73l_49
    Snippet: A pseudovirus bearing CoV S protein or VSV-G protein and a defective HIV-1 genome that expresses luciferase as reporter was produced in 293 T cells, as previously described (27) , and its titer was quantitated by using HIV-1 p24 ELISA. The pseudovirus was then used to infect target Huh-7 cells (or ACE2/293 T cells for pseudotyped SARS-CoV) (10 4 per well in 96-well plates) in the presence or absence of the test peptide at the indicated concentrat.....
    Document: A pseudovirus bearing CoV S protein or VSV-G protein and a defective HIV-1 genome that expresses luciferase as reporter was produced in 293 T cells, as previously described (27) , and its titer was quantitated by using HIV-1 p24 ELISA. The pseudovirus was then used to infect target Huh-7 cells (or ACE2/293 T cells for pseudotyped SARS-CoV) (10 4 per well in 96-well plates) in the presence or absence of the test peptide at the indicated concentration. Twelve hours after infection, culture medium was refreshed and then incubated for an additional 48 hours, followed by washing cells with PBS, lysing cells with lysis reagent (Promega), and transferring the cell lysates to 96-well Costar flat-bottom luminometer plates (Corning Costar) for the detection of relative light units using the Firefly Luciferase Assay Kit (Promega) and an Ultra 384 luminometer (Tecan).

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