A fast, robust and tunable synthetic gene oscillator - The BioCircuits ...

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doi: 10.1038/nature07389 SUPPLEMENTARY INFORMATION 400 Fluorescence (AU) 00 60 120 180 Time (min) 400 Fluorescence (AU) 00 400 60 120 180 Time (min) Fluorescence (AU) 00 60 120 180 Time (min) 400 Fluorescence (AU) 00 60 120 180 Time (min) 400 Fluorescence (AU) 00 60 120 180 Time (min) 240 240 240 240 240 400 Fluorescence (AU) 400 Fluorescence (AU) 00 60 120 180 Time (min) 400 Fluorescence (AU) 00 400 60 120 180 Time (min) Fluorescence (AU) 00 60 120 180 Time (min) 400 00 60 120 180 Time (min) 00 60 120 180 Time (min) Supplementary Figure 3: An example of unsmoothed fluorescence trajectories. These trajectories were taken from a timelapse microscopy run at 0.7% arabinose and 0.1 mM IPTG. Each trajectory is a single cell undergoing division and oscillation. The arbitrary fluorescence units on the y-axes are equivalent. www.nature.com/nature 7 Fluorescence (AU) 240 240 240 240 240
doi: 10.1038/nature07389 SUPPLEMENTARY INFORMATION Osc. period (min) 50 40 30 200 60 120 180 Duration of imaging (min) Supplementary Figure 4: The effect of continued growth on oscillatory period. JS011 cells were grown in microfluidic devices for long-term single-cell imaging under inducing conditions. Each point is an average of oscillation periods from several cells at the same point in time and the same conditions (black squares, 0 mM IPTG; purple diamonds, 0.25 mM; blue triangles, 0.5 mM; green circles, 0.75 mM; green crosses, 1 mM; yellow circles, 2 mM; orange triangles, 5 mM; red diamonds, 10 mM). Data analysis was performed using custom-written matlab scripts. IPTG (mM) oscillating cells dim cells bright cells dead cells 0 129 0 0 2 0.25 149 0 0 0 0.5 209 0 0 0 0.75 331 0 0 0 1 129 0 0 2 2 231 0 2 2 5 233 0 0 0 10 155 0 0 0 total 1566 0 2 6 (99.5%) (0.1%) (0.4%) Supplementary Table 1: The oscillator circuit is functional in almost every cell observed with single-cell microscopy. The experiments shown in Supplementary Movies 1–8 were examined, and single cells were identified. Each cell was tracked by eye and classified into one of four groups: oscillating, never fluorescent, always fluorescent (with saturating levels of fluorescence), and dead (with abnormal morphology, usually swollen or burst, and no fluorescence). www.nature.com/nature 8
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