Difference between revisions of "Part:BBa K2273003"
(8 intermediate revisions by the same user not shown) | |||
Line 10: | Line 10: | ||
</figure> | </figure> | ||
<p align="justify"> | <p align="justify"> | ||
− | The development of genetic competence in <i>Bacillus | + | The development of genetic competence in <i>Bacillus subtilis</i> is stimulated by quorum sensing (Figure 1). <i>B. subtilis</i> constantly secretes the <b>ComX pheromone</b>, a 9- to 10-amino acid oligopeptide, as a signalling molecule <b>(a)</b>. By rising cell-density, the ComX-concentration in the surrounding medium increases until it reaches a threshold and activates ComP, a membrane-spanning protein kinase <b>(b)</b>. |
The kinase reacts to the accumulation of ComX by phosphorylating the response regulator ComA <b>(c)</b> which then works as a transcription factor by binding to several promoters and enhancing their activity <b>(d)</b>. | The kinase reacts to the accumulation of ComX by phosphorylating the response regulator ComA <b>(c)</b> which then works as a transcription factor by binding to several promoters and enhancing their activity <b>(d)</b>. | ||
<a target="_blank" href="https://www.ncbi.nlm.nih.gov/pubmed/12576575">(Hamoen <i>et al.</i>)</a><br> | <a target="_blank" href="https://www.ncbi.nlm.nih.gov/pubmed/12576575">(Hamoen <i>et al.</i>)</a><br> | ||
Line 17: | Line 17: | ||
</figure> | </figure> | ||
<p> | <p> | ||
− | <br> | + | <br>We evaluated the influence of an overproduction of the ComX pheromone on the promoters that are listed below. |
<table> | <table> | ||
<tr><td>P<sub><i>srfA</i></sub></td><td><a href="https://parts.igem.org/Part:BBa_K2273000">BBa_K2273000</a></td></tr> | <tr><td>P<sub><i>srfA</i></sub></td><td><a href="https://parts.igem.org/Part:BBa_K2273000">BBa_K2273000</a></td></tr> | ||
<tr><td>P<sub><i>rapA</i></sub></td><td><a href="https://parts.igem.org/Part:BBa_K2273001">BBa_K2273001</a></td></tr> | <tr><td>P<sub><i>rapA</i></sub></td><td><a href="https://parts.igem.org/Part:BBa_K2273001">BBa_K2273001</a></td></tr> | ||
<tr><td>P<sub><i>rapF</i></sub></td><td><a href="https://parts.igem.org/Part:BBa_K2273002">BBa_K2273002</a></td></tr> | <tr><td>P<sub><i>rapF</i></sub></td><td><a href="https://parts.igem.org/Part:BBa_K2273002">BBa_K2273002</a></td></tr> | ||
− | |||
− | |||
</table> | </table> | ||
<br></p> | <br></p> | ||
+ | |||
<p> | <p> | ||
− | Especially for P<sub><i>srfA</i></sub> we could show ComX-dependence. | + | Especially for P<sub><i>srfA</i></sub> we could show ComX-dependence. By using an inducible copy of <i>comX</i> we could compensate <i>comX</i>-deficiency as well (Figure 2). |
</p> | </p> | ||
Line 34: | Line 33: | ||
<img src="https://static.igem.org/mediawiki/parts/4/41/T--TU_Dresden--P_Communication_comX.png" width="100%" > | <img src="https://static.igem.org/mediawiki/parts/4/41/T--TU_Dresden--P_Communication_comX.png" width="100%" > | ||
− | <figcaption><p align="justify"><b>Figure | + | <figcaption><p align="justify"><b>Figure 2: Growth (A) and promoter activity of various promoters (B) in dependence of xylose-induced ComX-production.</b> Wild type (WT, pink and light blue) and <i>comX</i>-knockout strains (<i>ΔcomX</i>, black and dark blue) contain a promoter-<i>luxABCDE</i> fusion as well as an inducible copy of <i>comX</i>. After 1 hour of incubation (dashed line) the cultures were induced with 1% xylose (light blue and black) or with distilled water (dH2O, pink and dark blue).</p></figcation> |
</figure> | </figure> | ||
<p align="justify"> | <p align="justify"> | ||
<b>The part was created and used by <a target="_blank" href="http://2017.igem.org/Team:TU_Dresden/Project/Communication">TU_Dresden 2017</a>.</b> All assays have been conducted with <i>B. subtilis</i> W168 carrying the promoter fused to the <i>comX</i> upstream a xylose-inducible promoter (P<sub><i>xylA</i></sub>). | <b>The part was created and used by <a target="_blank" href="http://2017.igem.org/Team:TU_Dresden/Project/Communication">TU_Dresden 2017</a>.</b> All assays have been conducted with <i>B. subtilis</i> W168 carrying the promoter fused to the <i>comX</i> upstream a xylose-inducible promoter (P<sub><i>xylA</i></sub>). | ||
+ | </html> | ||
− | Sequence and Features</span> | + | <span class='h3bb'>Sequence and Features</span> |
<partinfo>BBa_K2273003 SequenceAndFeatures</partinfo> | <partinfo>BBa_K2273003 SequenceAndFeatures</partinfo> | ||
Latest revision as of 21:03, 31 October 2017
ComX pheromone for competence development in Bacillus subtilis
Usage and Biology
We evaluated the influence of an overproduction of the ComX pheromone on the promoters that are listed below.
PsrfA | BBa_K2273000 |
PrapA | BBa_K2273001 |
PrapF | BBa_K2273002 |
Especially for PsrfA we could show ComX-dependence. By using an inducible copy of comX we could compensate comX-deficiency as well (Figure 2).
Figure 2: Growth (A) and promoter activity of various promoters (B) in dependence of xylose-induced ComX-production. Wild type (WT, pink and light blue) and comX-knockout strains (ΔcomX, black and dark blue) contain a promoter-luxABCDE fusion as well as an inducible copy of comX. After 1 hour of incubation (dashed line) the cultures were induced with 1% xylose (light blue and black) or with distilled water (dH2O, pink and dark blue).
The part was created and used by TU_Dresden 2017. All assays have been conducted with B. subtilis W168 carrying the promoter fused to the comX upstream a xylose-inducible promoter (PxylA).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]