Real-Time Single-Molecule Observation of Green Fluorescent Protein Synthesis by Immobilized Ribosomes

The dynamics of full protein synthesis and the co-translational folding processes are not fully understood. We have developed a novel method, using a combination of ribosome display and single-molecule techniques, for monitoring the synthesis, co-translational folding, and maturation of a complete polypeptide chain at the single-molecule level. This method enabled us to observe the appearance of green fluorescent protein fluorescence after de novo synthesis of the complete protein. Here, we provide the information necessary to reproduce this method, which will be valuable in revealing the dynamics of the co-translational folding and maturation of nascent polypeptides.