Figure 1

Regenerating the PURE system from genes. (A) Schematic of the basic biochemical reactions in the PURE system. The set of reactions is broken down in four main modules. The fuel molecule is creatine phosphate. Hydrolysis of pyrophosphate (PPi) into phosphate (P) is catalyzed by a pyrophosphatase. (B) Macromolecules constituting the PURE system, plus the tRNA- and rRNA-modifying enzymes that will eventually have to be synthesized in vitro for complete PURE system biogenesis. Note that 22 proteins constitute the 20 aaRSs. (C) Schematic map of the pTFM1 plasmid encoding all aminoacyl-tRNA synthetases (aaRSs) and translation factors (TFs), except EF-Tu. Adapted from Ref.¹³. Other abbreviations: AAs amino acids, AAtRNAs aminoacyl-tRNAs, aaRSs aminoacyl-tRNA synthetases, RNA pol RNA polymerase, Ribo ribosome, rProt ribosomal protein.