Results: 4

1.
Fig. 2.

Fig. 2. From: Spatiotemporal patterns and transcription kinetics of induced RNA in single bacterial cells.

RNA kinetics in single E. coli cells upon induction with ATc. (A, B) Time course of total fluorescence in individual E. coli cells in two representative experiments. The t = 0 time is defined as the moment of ATc addition to cells. Measurements are taken at 2-min intervals. (C) Control measurements using cells with expression of full-size EGFP (13). The fluorescence excitation intensity in the control experiments is identical to the intensity used in experiments shown in (A) and (B), but the emission intensity in (C) is attenuated 10-fold.

Maria Valencia-Burton, et al. Proc Natl Acad Sci U S A. 2009 September 22;106(38):16399-16404.
2.
Fig. 4.

Fig. 4. From: Spatiotemporal patterns and transcription kinetics of induced RNA in single bacterial cells.

Lateral fluorescence distribution in bacterial cells expressing RNP complexes is principally different from the random distribution of EGFP in E. coli cells. (A) Examples of cells expressing full-size EGFP (Left) and RNP complex (Right), and schematic illustration of the lateral fluorescence profiling approximating the cell as rods. (B) Typical examples of lateral fluorescence profiles in cells expressing EGFP (Left) and RNP complex (Right). The two lines in each panel represent the fluorescence intensity on the left and right sides of the rods. Insets show the correlation coefficient and standard deviation, where n is the number of the replicate measurements of one cell (for more examples, see Fig. S10). (C) Histogram of the correlation coefficients for 30 cells expressing EGFP (blue) and 31 cells expressing RNP (red) measured as in B, showing clear differences between the full-size EGFP-expressing cells and the RNP-expressing cells.

Maria Valencia-Burton, et al. Proc Natl Acad Sci U S A. 2009 September 22;106(38):16399-16404.
3.
Fig. 1.

Fig. 1. From: Spatiotemporal patterns and transcription kinetics of induced RNA in single bacterial cells.

RNA-inducible system to study transcription kinetics in live E. coli cells. (A) Experimental design. (B) A plasmid pMB133, expressing RNA tagged with the eIF4A-interacting aptamer under the control of TetR. TetR, expressed from the second T7 promoter, represses expression of RNA tagged with aptamer. Anhydrotetracycline (ATc) is required to relieve TetR repression and to induce RNA synthesis. (C) Plasmid expressing two protein fusions from the two T7/lacO promoters. (D) Flow cytometry analysis of E. coli cells transformed with the plasmids MB133 and MB33 after induction of RNA synthesis with anhydrotetracycline (ATc). (E) RNA concentration after induction with ATc as determined by competitive PCR (for details see ref. 13).

Maria Valencia-Burton, et al. Proc Natl Acad Sci U S A. 2009 September 22;106(38):16399-16404.
4.
Fig. 3.

Fig. 3. From: Spatiotemporal patterns and transcription kinetics of induced RNA in single bacterial cells.

Time-lapse fluorescent images of live E. coli cells expressing RNA. Time after ATc induction is given in minutes in each image. Bars represent 2 μm. (A) In this E. coli cell, fluorescence is localized mainly along the edges of the cells (best visible in the time window 55–85 min). Focal points at the mid-cell and quarter-cell positions appear ≈100 min after RNA induction with ATc (see Fig. S3 for the full set of time-lapse images). (B) In this E. coli cell, the peripheral distribution of fluorescence is clearly visible in images in the time window 36–78 min. Mid-cell focal accumulation appears ≈75 min after RNA induction with ATc (see also Fig. S4). (C) A small E. coli cell showing nonuniform lateral fluorescence distribution. The dimmer fluorescence in the middle of the cell is best visible in the time window 85–109 min (see also Fig. S5). (D) A cell displaying polar accumulation of fluorescence (see also Fig. S6). (E) An E. coli cell displaying time-dependent fluorescent accumulation at a pole, which disappears later in the cycle and reappears at a different location (see also Fig. S7). (F) Examples of E. coli cells with fluorescence accumulations at the edges of the cell, likely indicators of helical structures, and the cells with visible diffuse helical structures slanted to the long cell axis.

Maria Valencia-Burton, et al. Proc Natl Acad Sci U S A. 2009 September 22;106(38):16399-16404.

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