Figure 4.

Binding of MobMN199 to the PctII promoter on single-stranded DNA. (A) Sequence alignment of the minimal oriT sequence (IR1+8; coordinates 3570–3595) and the PctII promoter region spanning coordinates 863–840 (positions −27 to −4). Arrows illustrate inverted repeats. The −10 box of the PctII promoter is indicated. (B) Stem–loop structure predictions for oligonucleotides IR1+8 and PctII. (C) Competitive EMSA. Different concentrations of non-labelled competitor DNA (oligonucleotides PctII and IR1+8) were mixed with Cy5-labelled oligonucleotide IR1+8 (2 nM). Then, protein MobMN199 (80 nM) was added to the reaction mixtures. Samples were loaded onto a native polyacrylamide (10%) gel. The positions of free DNA (F) and bound DNA (C) are indicated.