Reconstitution of 30S ribosomal subunits in vitro using ribosome biogenesis factors

The effects of the biogenesis factors. (A) The effect of each biogenesis factor or their combination was assayed using a PURE direct assay. (B) Analysis of the effect of Era in the two-step PURE direct assay. The 30S subunit assembly reaction was performed for 60 min at 37°C without translation mixtures, including components such as native 50S subunits, poly(U), ¹⁴C-phenylalanine, and elongation factors. After the assembly reaction, the level of polyphenylalanine synthesis was monitored 30 and 60 min after addition of the translation mixture.