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. 2016 Jan 17;44(4):1514–1524. doi: 10.1093/nar/gkw007

Figure 5.

Figure 5.

Gene expression correlates with TCDS level of neighboring genes. (A) Scheme of the reporter gene arrangement. The tufB promoter was chosen due to its expression strength for reliable detection of the signal and the clear correlation of negative neighboring TCDS and gene expression. In the construct designed in the native locus of tufB the coding sequence of tufB is replaced by the reporter gene venus encoding a fluorescent protein. Upstream of the tufB promoter the tet promoter coupled to mCherry reporter gene is inserted. (B) Temporal expression of tufB in wild-type Escherichia coli (21) and TCDS derived from the expression and orientation of neighboring genes within 10 kb. (C) Total expression of tetP (mCherry) at 24 h after induction with different concentrations of Anhydrotetracyclin. Abscissa: no repressor (cells without repressor plasmid), induction with decreasing levels (from left to right) of Anhydrotetracyclin and no tetP insert (the construct without the tetP insert). Ordinate: total fluorescence of mCherry after 24 h. (D) The same as in (C) with venus total fluorescence measured 24 h after induction.