Figure 2
In vitro production of 32 translation factor proteins from a single plasmid. (A) Reaction scheme for pTFM1 expression in the PURE system. AAs amino acids, TFs translation factors, FMT methionyl-tRNA-formyltransferase, IF initiation factor, EF elongation factor, RF release factor, RRF ribosome recycling factor. (B,C) Relative expression levels of proteins expressed from pTFM1 with PUREfrex2.0 (B) and PURExpress (C) after 5 h incubation at 37 °C. Ratios of 15N-labeled peptides (newly synthesized proteins) to 14N peptides (original proteins) are displayed for all measured peptides; for proteins with multiple measured peptides, the peptides are ordered from N- to C-terminus (top to bottom). Error bars denote ± 1 standard deviation from triplicate experiments. Results from a different batch of PUREfrex2.0 are shown in Supplementary Fig. 2. (D) Specific peptides measured in (B) and (C).
