Fig. 4
Investigation of CrCD protein accumulation in C. reinhardtii tCI cell lines upon varying the number of internal TGA codons and/or induction temperature. a Cell lines with 2, 4 or 6 internal TGA codons in the CD gene were pre-cultured at 30 °C then shifted to 18, 25 or 30 °C for induction. CrCD protein accumulation was measured by anti-HA immunoblot after 72 h. Equal culture volumes were used for each immunoblot sample, then anti-HA band intensity values were divided by the optical density (750 nm) of that culture to plot relative ‘per cell’ CrCD. Each bar shows the mean value for two cultures. b The CD/4* cell line was grown in four identical flask cultures at 35 °C to a high cell density (OD750 = 2.1) then induced at 15, 18, 20 or 25 °C using Algem photobioreactors. CrCD accumulation was followed for 72 h after induction. Samples were equalised according to optical density at 750 nm and analysed by SDS-PAGE and anti-HA immunoblotting. Error bars (on all data points) show ± SD for two samples taken from the same flask at each timepoint