ParB phylogeny and <i>parS</i> sequences of the Par systems used in this study.

<p>ParB Phylogeny was obtained using the multiple sequence alignment webPRANK of the European Bioinformatic Institute (<a href="http://www.ebi.ac.uk/goldman-srv/webprank/" target="_blank">http://www.ebi.ac.uk/goldman-srv/webprank/</a>). The Par systems are from the following replicons: c1, chromosome 1 of <i>Burkholderia cenocepacia</i> J2315; G4, plasmid 2 of <i>Burkholderia vietnamiensis</i> G4; 12D, plasmid 1 of <i>Ralstonia pickettii</i> 12D; c3, chromosome 3 of <i>B</i>. <i>cenocepacia</i> J2315; 12J, plasmid integrated in the chromosome of <i>Ralstonia pickettii</i> 12J; pBC, plasmid pBC of <i>B</i>. <i>cenocepacia</i> J2315. Families correspond to those previously established based on a wider collection of Par systems. All <i>parS</i> sequences form a perfect palindrome (indicated by the inverted arrows with specific colours for specific sequences), except <i>parS</i>12J which displays non complementary bases (boldface).</p

Binding scores for palindromic <i>parS</i> and derivatives.

<p>(A) ParB c1 <i>B</i>. <i>cenocepacia;</i> (B) ParB c3 <i>B</i>. <i>cenocepacia;</i> (C) ParB plasmid pBC <i>B</i>. <i>cenocepacia</i>; (D) ParB plasmid 1 <i>Ralstonia picketti</i> 12D; (E) ParB plasmid 2 <i>Burkholderia vietnamiensis</i> G4. Binding scores correspond to 100-fold the VR value at the end of the SPRi injection. The average scores for the wild-type (wt) and random (rdm) sequence are represented as lines, green and red respectively. Average scores for the half-<i>parS</i> sequences are shown as orange lines. For double symmetric substitutions, the score is directly pointed on the graph as the base substituting the wild-type one indicated below on the x-axis. Changes are indicated for one arm only but correspond to symmetrical changes (<i>e</i>.<i>g</i> the change T1C in <i>parS</i>c1 means T1C and A16G). For each change statistical analysis of the BS were performed; the substitutions displaying a significantly different BS relative to that of non specific DNA are wrote in red.</p

Experimental design.

<p><b>(A)</b> Structures of the 5’-thiol-labelled 57-mer single strand oligonucleotides carrying carrying <i>parS</i>, exemplified for <i>parS</i>c1. The <i>parS</i> sequence forms by the annealing of two inverted repeats of 26 bases (arrowed boxes). The upper structure shows the wild-type (wt) 16 bp palindrome of <i>parS</i>c1 by two arrowed black lines and with bp positions numbered. Three sequences modified from <i>parS</i>c1 are indicated below: randomly modified (rdm), one arm modified (half), and the double symmetric change T1C-A16G, (changed sequence is written in red) <b>(B)</b> Typical SPRi image captured at the end of injection (240 sec) of 10μg/ml of crude extract enriched in ParB c1. Each DNA probes was distributed in duplicate in two different localizations, as shown by white rectangles with <i>parS</i> wild type sequence (c1 wt) and white dotted rectangles with randomized sequence (c1 rdm). <b>(C)</b> Examples of kinetics curves for the six ParB-<i>parS</i> systems. For each ParB, the reflectivity variations according to time were observed with the wild-type <i>parS</i> (green curves) and the random <i>parS</i> (red curves). Various concentrations of crude extracts were injected: 10 (dotted lines), 20 (dashed lines) 40 μg/ml (full lines) for c1; 20 (dotted lines), 40 (dashed lines) and 80 μg/ml (full lines) for pBC, c3, 12D, 12J and G4.</p

Binding scores of ParB12 J for <i>parS</i>c3 non-<i>parS</i>c3 sequences and derivatives.

<p>Blue lines correspond to the BS of the palindromic sequence <i>parS</i>c3 (A) and non <i>parS</i>c3 (B). Legend is the same as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0177056#pone.0177056.g003" target="_blank">Fig 3</a>.</p

Additional file 1: of Inactivation of spores by electric arcs

Determination of density of untreated spores. Successive spore dilutions of 1 hundred were shown after spread and incubation overnight at 37°C. A bacterial mat was observed for the 1/1 dilution (a). A very high density of colony was visualized in the 1/100 dilution (b). 222 colony was counted in the 1/10 000 dilution (c). With a petri dish surface of 63.6 cm², we calculated a spore density of 3.5 104 spore by cm² in the 1/1 dilution. (DOCX 2.59 MB