Fig. 1: Diagram of in vitro ribosome synthesis and evolution (RISE).
From: In vitro ribosome synthesis and evolution through ribosome display
Starting with a T7-promoted rDNA operon, mutations are introduced to generate an rDNA operon library. The rDNA operon library is then used in an iSAT reaction to assemble a library of ribosomes. Functional ribosomes then translate selective peptides and stall to form ternary complexes, allowing for selection of the peptides and, thus, isolation of functional ribosomes. The rRNA of the functional ribosomes is then purified and reverse transcribed into cDNA for sequencing and reinsertion into the rDNA operon.
