Fig. 5

Cold-inducible synthesis of cis-abienol synthase, TPS4. a Design of chloroplast expression constructs. The TPS4 genes include a C-terminal HA tag for detection. b Immunoblot using anti-HA antibodies to detect TPS4 protein in C. reinhardtii tCI cell lines induced at 18 °C. Samples were equalised according to the culture optical density at 750 nm before loading. The first three lanes show a constitutive TPS4 control cell line with no internal TGA codons. c Anti-HA immunoblot to compare induction temperatures of 18 and 25 °C in C. reinhardtii. Samples were equalised according to the culture optical density at 750 nm before loading. A strain synthesising CrCD (right lane) is included as a negative control for the TPS4 band. d Quantification of the band intensities in part c, showing the TPS4 induction time-course at two temperatures. e Anti-HA immunoblot to detect TPS4 protein in E. coli DH5α containing various TPS4 chloroplast expression plasmids. Total protein stain for this blot is shown in Additional file 1: Figure S6. The incubation temperature is given above each lane