Figure 2 | Scientific Reports

Figure 2

From: Enhancing the performance of a mutant pyrrolysyl-tRNA synthetase to create a highly versatile eukaryotic cell-free protein synthesis tool

Figure 2

Detection of amber-suppression efficiency in cell-free protein synthesis. (a) PylRS-AF was synthesized in cell-free reactions based on E.coli, wheat germ (WG), Sf21 and CHO in a CECF format. The supernatant of these reactions was supplemented to a CHO batch based cell-free reaction containing Adora2a-amb-Nluc as template. In presence of active PylRS the full length protein with C-terminal luciferase was obtained. Nluc activity was measured after three hours synthesis reaction. The maximal possible PylRS-AF concentration and equal enzyme concentrations (80 nM) were added to the batch based reaction. (b) The same supernatant described in (a) was supplemented to a CHO based cell-free reaction containing Adora2a-Nluc to determine the effect of supernatants on the translation reaction. Nluc activity was measured after three hours synthesis reaction. Measurements were performed in duplicate. Data are shown as mean ± SD. Independent experiments were performed two times and can be found in the Suppl. Fig. S11.

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