Figure 2.

Cell-free syntheses yield Pseudomonas aeruginosa PilD and Methanococcus voltae FlaK active against cognate substrates. The final pellet from a small-scale translation reaction was resuspended in 25 mmol/L HEPES, pH 6.5 then used in these activity assays. (A) PilD activity: pellets of the translation reactions of the empty vector (EV); PilD; PilD + PilA cotranslation and PilA were separated by SDS-PAGE, followed by Western blot analysis using PilA-specific antiserum. (B) FlaK activity: pellets of the translation reactions of the EV; FlaB2 substrate, FlaK enzyme, and FlaK + FlaB2 cotranslation were separated by SDS-PAGE, followed by Western blot analysis using FlaB2-specific antibodies.