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. 2019 Oct 10;14(10):e0222964. doi: 10.1371/journal.pone.0222964

Fig 6. Application of devices to tRNA-tRNA FRET.

Fig 6

(a) Schematic cartoon of ribosomes immobilized in an Al ZMW well via biotinylated mRNA, with FRET between Phe-tRNAPhe(Cy5) in the ribosomal P-site and Val-tRNAVal(Cy3) in the A-site. The ribosomes were illuminated by 532 nm laser light, in the presence of 500 nM Phe-tRNAPhe(Cy5)·GTP·elongation factor-Tu (ternary complex) in solution. (b) Example FRET trace from a ribosome transitioning from the low FRET hybrid state to the high FRET classical PRE-translocation state before photobleaching of Cy5 at 7 s followed by photobleaching of Cy3 at 15 s.