Development of a Heat-Shock Inducible Gene Expression System in the Red Alga Cyanidioschyzon merolae
Figure 3
Transcriptional activities of the upstream region of CMJ101C orf before and after heat shock treatment in transiently transformed cells.
Several different lengths of the upstream region of CMJ101C orf were respectively fused with sfGFP orf. The plasmids were transiently transformed into C. merolae wild-type cells. After transformation, cells were cultured at 36°C for 1 day and then shifted to 50°C for 1 h. Expression of the GFP protein was examined by fluorescence microscopy and immunoblotting. (A) Schematic diagram of the plasmids used. (B and C) The percentage of the cells with GFP fluorescence was compared before and after the 1-h 50°C heat shock treatment. The red is the autofluorescence of chlorophyll. The error bars represent the standard deviation calculated from three individual experiments (n = 3). 100 cells were counted for each experiment. The scale bar is 10 µm. (D) Immunoblotting with the anti-GFP antibody before (36°C) and after (50°C) the heat shock treatment. CBB staining of the PVDF membrane is shown as a loading control.