Figure 6. Western Blot analysis of leaf and seeds samples of transgenic Geudertheimer plants (100 µg TSP); (A) Samples from stable transformed plants; E: 5 ng of recombinant C5a produced in E. coli; Control: leaf and seed samples from wild-type tobacco.
C5aVac: leaf and seed samples of the T2 individual C5aVac 19-13-15, expressing the vacuolar variant of C5a; (B) Samples from N. benthamiana at 6 dpi after Agroinfiltration; control: N. benthamiana transfected with the empty vector pICH29912; C5aVac: N. benthamiana transfected with pICH29912-C5aVac; E: 5 ng of recombinant C5a produced in E. coli; (C) Verification of C5a biological activity using crude leaf extracts from transgenic tobacco and the near isogenic variant (wildtype) compared to a commercial standard C5a produced in E. coli. Enzyme release was measured by substrate conversion following exposure to different dilutions of the lead extracts, with the concentration of C5a determined by ELISA.