Figure 1
Construction and characterization of the LHCSR1-luciferase reporter strain. (A) Construction map for introducing the LHCSR1-luciferase reporter gene into Chlamydomonas cells. PAR and PLHCSR1 indicate the HSP70A-RBCS2 tandem promoter and LHCSR1 promoter regions, respectively. TR and TLHCSR1 indicate the terminator regions of RBCS2 and LHCSR1, respectively. The LHCSR1 gene, including exons and introns, is represented by red boxes. The codon-optimized firefly luciferase (LUCnc) gene and paromomycin resistance gene (AphVIII) are represented by the yellow box and black box, respectively. (B) Bioluminescence activity of the reporter strain under various colored lights. The intensities of the fluorescent light with UV and the blue (450 nm), green (530 nm) and red (660 nm) monochromatic LED lights were 30 μmol/m2/s. Bioluminescence activities of the light treated cells were normalized to the bioluminescence activity under LL. Data represent the mean ± SEM of three biological replicates. Only UV-treated cells showed a significant increase in activity (one-way ANOVA, P < 0.0001). (C) Immunoblot analysis of LHCSR1 and LHCSR1-luciferase fusion protein in the reporter strain obtained in (B) using a specific antibody against LHCSR1. The signals were obtained from the same blot at different exposure times (short and long exposure for LHCSR1 and LHCSR1-luciferase signals, respectively). Raw images of the blot are shown in the supplemental information (Supplementary Fig. S2). The samples illustrated here are representative of three biological replicates.
