Figure 4.
Expression of the in‐frame gene 3CLpro/S‐II/Luc for the cis‐cleavage analysis. (A–F) Microscopical analysis of the Vero cells with bright field (left) and fluorescent light (right). Vero cells (B–F) transfected with pEGFP‐N1 plus pcDNA3.1, Pro/S‐II, Pro/S‐II/Luc, Pro/S‐II(L → A)/Luc or Pro/S‐II(Q → A)/Luc were compared to the un‐transfected cells (A). (G) Equal amounts (100 × g) of cell lysates were used to determine the Luc activity using the dual Luciferase Reporter Assay System. The reference cells transfected with pcDNA3.1 or Pro/S‐II were used to subtract background noise. Relative Luc activity in the test cells transfected with Pro/S‐II/Luc, Pro/S‐II(L → A) or Pro/S‐II(Q → A)/Luc was determined as the value of light unitstest‐reference.