Maximal fluorescence values of the mCherry protein reached during prolonged growth in CM-G medium normalized by the OD 600 nm in single and double transformants bacteria, harbouring the functional derivatives pTLR1-P_X (dark blue), pTLR1-P_A (purple), pTLR1-P₂ (sky blue) pTLR1-P_D1 (orange), and pTLR1d used as negative control (red).

<p>Standard deviation bars for the different replicates are included.</p

Oligonucleotides used in this study.

<p>*Restriction enzyme sites are depicted in italics.</p

Figure 1

<p>(A) Schematic representation of the <i>as-48</i> (black) and <i>bac21</i> (grey) gene clusters. Solid black arrows represent the proposed promoter regions and dotted arrows indicate the mRNAs detected by Fernandez <i>et al.... </i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0090603#pone.0090603-Fernndez1" target="_blank">[10]</a>, Díaz <i>et al.... </i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0090603#pone.0090603-Daz1" target="_blank">[5]</a> and Martínez-Bueno <i>et al.... </i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0090603#pone.0090603-MartnezBueno2" target="_blank">[9]</a>. Solid grey arrows represent the promoter regions proposed by Tomita <i>et al.... </i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0090603#pone.0090603-Tomita1" target="_blank">[6]</a>. (B) Promoters identified <i>in silico</i> (dashed arrows) and their location according to AS-48 nomenclature (Genebank KJ146793 and Y12234.1): P_A (nt 1105-nt 1396), Pc (nt 2129-nt 2477), P₂₍₂₎ (nt 2788-nt 3163), P₂₍₁₎ (nt 2788-nt 3010), P_D1 (nt 3721-nt 4160), P₃₍₁₎ (nt 4353-nt 4544) and P₃₍₂₎ (nt 4188-nt 4544). Predicted terminators according to BPROM <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0090603#pone.0090603-Solovyev1" target="_blank">[32]</a> in <i>as-48</i> gene cluster are pointed with a <b>T</b>.</p

Expression of mCherry normalized by the OD 600-derivatives in <i>E. coli</i> TOP10, <i>L. lactis</i> LM2301 and <i>E. faecalis</i> JH2-2 during prolonged growth in CM-G medium.

<p>Fluorescence emission of mCherry was recorded at 620-P_X (dark blue), pLTR1-P_A (purple), pLTR1-P₂₍₂₎ (sky blue), pLTR1-P_D1 (orange) and pTLR1d (red) used as negative control. Standard deviation bars for the different replicates are included.</p

Strains and plasmids used in this study. Cm^R chloramphenicol resistant, Em^R erythromycin resistant.

<p>Strains and plasmids used in this study. Cm^R chloramphenicol resistant, Em^R erythromycin resistant.</p