Difference between revisions of "Part:BBa K2273000"
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===Usage and Biology===
 
===Usage and Biology===
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<html><figure>deedfewofkrweokewfqwkw<figure style="width:50%;float:left;"><img src="https://static.igem.org/mediawiki/parts/1/11/T--TU_Dresden--P_Communication_Psrf_Media.png" width="100%"><figcaption><b>Figure 2: Growth (A) and promoter activity of P<sub><i><b>srfA</b></i></sub> (B) in different media.</b> Wild type (WT, pink) and <i>comX</i>-knockout strains (<i>ΔcomX</i>, dark blue) contain a P<sub><i>srfA</i></sub> <i>luxABCDE</i> fusion. </figcation></figure></figure></html>
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<html><figure>The well-studied regulatory system for competence development in <i>B. subtilis</i> provided a genetic set-up based on quorum sensing that we used as a proof of principle <a target="_blank" href="https://www.ncbi.nlm.nih.gov/pubmed/12576575">(Hamoen <i>et al.</i>, 2003)</a> (Figure 1).
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<i>B. subtilis</i> constantly secretes the ComX pheromone, 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>.
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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>
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<a target="_blank" href="https://www.ncbi.nlm.nih.gov/pubmed/26582911">[5</a>,<a target="_blank" href="https://www.ncbi.nlm.nih.gov/pubmed/18585392">6]</a>.</p>
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The most important promoter regulated by ComA and involved in competence development is the promoter of the <i>srfA</i> operon. This operon contains not only genes for the production of the antibiotic surfactin <b>(e)</b> but also for ComS <b>(f)</b>, another small peptide that prevents the degradation of the autoregulated transcription factor ComK <b>(g)</b><figure style="width:50%;float:left;"><img src="https://static.igem.org/mediawiki/parts/1/11/T--TU_Dresden--P_Communication_Psrf_Media.png" width="100%"><figcaption><b>Figure 2: Growth (A) and promoter activity of P<sub><i><b>srfA</b></i></sub> (B) in different media.</b> Wild type (WT, pink) and <i>comX</i>-knockout strains (<i>ΔcomX</i>, dark blue) contain a P<sub><i>srfA</i></sub> <i>luxABCDE</i> fusion. </figcation></figure></figure></html>
  
 
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Revision as of 19:06, 30 October 2017


PsrfA: a ComA-dependent promoter of Bacillus subtilis

Figure 1: The regulatory system for competence development in Bacillus subtilis. The development of genetic competence is stimulated by quorum sensing due to the secretion of the ComX pheromone and its accumulation in the surrounding medium. (Modified from: Jung et al.)

Usage and Biology

The well-studied regulatory system for competence development in B. subtilis provided a genetic set-up based on quorum sensing that we used as a proof of principle (Hamoen et al., 2003) (Figure 1). B. subtilis constantly secretes the ComX pheromone, a 9- to 10-amino acid oligopeptide, as a signalling molecule (a). 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). The kinase reacts to the accumulation of ComX by phosphorylating the response regulator ComA (c) which then works as a transcription factor by binding to several promoters and enhancing their activity (d) [5,6].

The most important promoter regulated by ComA and involved in competence development is the promoter of the srfA operon. This operon contains not only genes for the production of the antibiotic surfactin (e) but also for ComS (f), another small peptide that prevents the degradation of the autoregulated transcription factor ComK (g)

Figure 2: Growth (A) and promoter activity of PsrfA (B) in different media. Wild type (WT, pink) and comX-knockout strains (ΔcomX, dark blue) contain a PsrfA luxABCDE fusion.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]