Add Coelenterazine-h v1

AddCoelenterazine-has the substrate (light-sensitive; prepare fresh on the day of use) to activate the full-length RenG luciferase and measure its maximum luminescence value. This serves as the relative standard for subsequent SLCA interaction signals. Since three configurations will be tested, record the strains and culture conditions to ensure consistency. Next, measure the luminescence intensity of the three SLCA constructs to compare their complementation efficiency and background signals, thereby establishing the optimal pairing of the SpyTag/SpyCatcher system and the positioning of the RenG fragments. SLCA assay 1:Full-length RenG fusion protein, serving as the maximum luminescence reference. SLCA assay 2:SpyTag + RenG (N-terminal) with SpyCatcher + RenG (C-terminal). SLCA assay 3:SpyTag + RenG (C-terminal) with SpyCatcher + RenG (N-terminal).