Fig. 1: Single-molecule tracking of ribosomal subunits.
From: Direct measurements of mRNA translation kinetics in living cells
a Site-specific labeling of ribosomal subunits was achieved by genetically fusing the HaloTag protein to a ribosomal protein (L1, L9, L19, or L25 of the 50S subunit, and S2 of the 30S subunit). L1 is not resolved in the shown structure, and hence not highlighted. b Site-specific labelling of only a subpopulation of ribosomal subunits was achieved by introducing the MS2 RNA aptamer into a surface exposed helix of ribosomal RNA (23S rRNA H98 or H101, 16S rRNA h6 or h44), to which a co-expressed MS2CP-HaloTag fusion protein binds. c Fluorescence time-lapse movies of cells were acquired, and diffusion trajectories of labeled ribosomal subunits were automatically detected in cells segmented based on phase-contrast images.
