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. Author manuscript; available in PMC: 2015 Jun 8.
Published in final edited form as: Nat Struct Mol Biol. 2010 Jun 20;17(7):793–800. doi: 10.1038/nsmb.1828

Figure 1. Intersubunit FRET monitors the elongation cycle at each codon during translation.

Figure 1

(a) Single-molecule translation assay. Cy5-labeled 50S subunits, ternary complex, and EF-G are delivered to surface-immobilized single PICs containing Cy3-labeled 30S subunits and illuminated at 532 nm; both Cy3 and Cy5 fluorescence are simultaneously detected. (b) Immobilization with an mRNA coding for 6 phenylalanines (6F) permits the observation of ribosome conformation during multiple rounds of elongation via the intersubunit FRET signal. The arrival of FRET corresponds to 50S subunit joining during initiation, and is followed by multiple cycles of high-low-high FRET, each reporting on ribosome unlocking and locking during one round of elongation.