Fig. 1
Screening for Fd-HydA expressing clones. C. reinhardtii hydA 1,2 double mutant cells were transformed with Fd-HydA either under a Hsp70-RbcS2 or b psaD promoter. The transformants were screened for gene expression using the H2 sensitive R. capsulatus screen. Chlorophyll fluorescence of the algae is shown in red whereas dark halation represents GFP produced by R. capsulatus upon H2 presence. The wt strain CC-124 (white circle) and hydA 1,2 double mutant (blue circle) clones were used as positive and negative controls, respectively. The highest Fd-HydA expressing clones (1D4 for Hsp70-RbcS2 and P6 for psaD) are circled with purple and yellow, respectively. c Immunoblot analysis for detection of Fd-HydA expression. Soluble proteins (75 µg of each) from 1D4 (lane 1) and P6 (lane 2) were loaded on 4-12 % Bis–Tris PAGE (Life Technologies) and probed with rabbit polyclonal HydA1/2 antibodies. C. reinhardtii Fd-HydA (10 ng) expressed heterologously and purified from E. coli (co-loaded with soluble protein from the hydA 1,2 mutant) was used as positive control (lane 4)