7 research outputs found
Fragmentation spectra of the ions of mature forms of SHP1299, SHP1555 and SHP1509.
<p>Fragmentation of the ions m/z 1018.56 (A) and m/z 564.28 (B) identified in the supernatant of cultures of <i>S. thermophilus</i> strain LMD-9. Fragmentation of the ions m/z 799.49 (C) identified in the supernatant of cultures of <i>S. agalactiae</i> strain NEM316 and m/z 872.5 (D) identified in the supernatant of cultures of <i>S</i>. <i>mutans</i> strain UA159. All ions were analyzed in the linear ion trap.</p
Plasmids used in this study.
a<p>Ts indicates that the plasmid encodes a thermosensitive RepA protein.</p>b<p>Km and Erm indicate resistance to kanamycin and erythromycin, respectively.</p
Growth and luciferase activities of strains containing P<i><sub>shp</sub></i>-<i>luxAB</i> fusions in various genetic backgrounds.
<p>Growth curves (OD<sub>600</sub>) are presented in gray and relative luciferase activities (RLU/OD<sub>600</sub>) in black. Growth and relative luciferase activities of derivatives of <i>S. thermophilus</i> strain LMD-9 grown in CDM and containing P<i><sub>shp</sub>-luxAB</i> fusions of the loci <i>shp/gbs1555</i> of <i>S. agalactiae</i> (A), <i>shp/</i>SMU.1509 of <i>S. mutans</i> (B) and <i>shp/ster_1299</i> of <i>S. thermophilus</i> strain LMD-9 (C). The genetic backgrounds are indicated as follows: (•) the <i>shp</i> and <i>rgg</i> genes of the locus tested and the <i>ami</i> gene <i>of S. thermophilus</i> are present (▴) the cognate <i>shp</i> gene of the locus studied is not present, (▪) the cognate <i>rgg</i> gene of the locus studied is not present and, (<b>×</b>) the <i>ami</i> genes of <i>S. thermophilus</i> are not present. Experiments were done at 30°C for the <i>shp/gbs1555</i> and the <i>shp/</i>SMU.1509 loci and at 42°C for the <i>shp/ster_1299</i> locus. Data shown are representative of three independent experiments.</p
Description of strains containing P<i><sub>shp</sub></i>-<i>luxAB</i> transcriptional fusions in various genetic backgrounds.
<p>These strains were constructed in <i>S. thermophilus</i> strain LMD-9 and used to study the expression of the <i>shp</i> genes of <i>S. agalactiae</i> strain NEM316 (<i>shp/gbs1555</i> locus) and <i>S. mutans</i> strain UA159 (<i>shp/</i>SMU.1509 locus) in the presence and absence of the corresponding <i>shp</i> and <i>rgg</i> genes and in the presence and absence of the <i>ami</i> genes of <i>S. thermophilus</i> strain LMD-9.</p
Bacterial strains used in this study.
a<p>Km and Erm indicate resistance to kanamycin and erythromycin, respectively.</p>b<p>Arrows indicate construction by transformation with chromosomal DNA or plasmid.</p>c<p><i>shp1299</i> is annotated <i>ster_1298</i> in Genbank.</p
Cross-complementation of the <i>shp/rgg</i> loci with synthetic SHP pheromones.
<p>Maximum relative luciferase activities of the reporter strains TIL1052 (<i>shp1299</i>::<i>erm blp</i>::P<i><sub>shp1299</sub>-luxAB aphA3</i>), TIL1200 (Δ<i>shp1358 blp</i>::P<i><sub>shp1358</sub>-luxAB</i>), TIL1382 (<i>blp</i>::<i>gbs1555</i>::P<i><sub>shp1555</sub></i>-<i>luxAB aphA3</i>) and TIL1384 (<i>blp</i>::SMU.1509::P<i><sub>shp1509</sub></i>-<i>luxAB aphA3)</i> grown in the absence (grey) or in the presence of synthetic SHP peptides added at the beginning of the culture to a concentration of 1 µM: EGIIVIVVG (green), DILIIVGG (red), DIIIIVGG (blue), ETIIIIGGG (purple), DIIIFPPFG (yellow). The legitimate SHP synthetic peptide associated to the locus studied is hatched in each case.</p
The <i>shp/rgg</i> loci used in this study.
a<p>Group number of the SHP-associated Rgg according to the classification described in Fleuchot <i>et al.</i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066042#pone.0066042-Fleuchot1" target="_blank">[21]</a>.</p>b<p>The <i>shp</i> gene is followed by the Genbank id of the <i>rgg</i> genes.</p>c<p>The <i>shp</i> genes are not annotated in Genbank but were identified using BactgeneSHOW <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066042#pone.0066042-Ibrahim2" target="_blank">[20]</a>, except for the <i>shp</i> gene associated with <i>ster_1299</i>, which is annotated <i>ster_1298</i> in the genome of <i>S. thermophilus</i> strain LMD-9. Consequently, all the <i>shp</i> gene products are indicated with the term “SHP” followed by the number of the cognate <i>rgg</i> gene in Genbank. To unify the nomenclature, the <i>ster_1298</i> gene product was renamed SHP1299.</p>d<p>The sequences of the synthetic peptides used in this study are underlined.</p