Bacterial strains and plasmids used in this study.

<p>Bacterial strains and plasmids used in this study.</p

Effect of overexpression of Fis on the level of transcription from the <i>lapF</i> promoter.

<p>β-Galactosidase (β-Gal) activity expressed from the <i>lapF</i> promoter-lacZ reporter constructs were measured in the wild-type strain PSm and the Fis-overexpressing strain F15 of <i>P. putida</i> grown for 18 hours in LB medium supplemented with 1 mM IPTG and without IPTG supplementation. Vertical bars denote 95% confidence intervals of means. Data of at least 8 independent measurements is shown. The length of the <i>lapF</i> promoter region inserted upstream of the <i>lacZ</i> reporter gene in different pBLKT constructs is shown above diagrams. The Fis-binding site Fis-F2 (gray box) and 5′ end of <i>lapF</i> mRNA F-I (arrow) are shown in drawings.</p

Gel shift assay of the Fis binding to the <i>lapF</i> promoter DNA containing the wild-type Fis binding site Fis-F2 and the mutated site Fis-F2-mut.

<p>2×10¹⁰ molecules of radioactively labelled PCR products containing Fis-F2 (lanes 1–10) and Fis-F2-mut site (lanes 11–20) were used for Fis binding. Fis was outcompeted from Fis-DNA complex with unlabelled PCR product containing the Fis binding site (LF2) and PCR product without Fis-binding site (RF1). Added unlabelled DNA was calculated in molecules. 0.46 µM Fis was used in each reaction mixture except mixtures without Fis in lanes 2 and 12.</p