Difference between revisions of "Part:BBa K1413023"
 
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bphR2 gene, from Pseudomonas pseudoalcaligens KF707, allows the transcription of bphR1 which activates the genes of degradation of biphenyls. In our project, genes of degradation was replaced by RFP gene to detect the compound.
 
bphR2 gene, from Pseudomonas pseudoalcaligens KF707, allows the transcription of bphR1 which activates the genes of degradation of biphenyls. In our project, genes of degradation was replaced by RFP gene to detect the compound.
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<br/><br/>In absence of PCBs, bphR2 (BBa_K1413021) is bound to bphR1 promoter which activate the transcription of RFP but in very low expression.  
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This part is composed by constitutive promoter <a href="https://parts.igem.org/Part:BBa_J23114">(BBa_J23114)</a>, RBS <a href="https://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, bphr2 <a href="https://parts.igem.org/Part:BBa_K1413021">(BBa_K1413021)</a> and terminator <a href="https://parts.igem.org/Part:BBa_B0015">(BBa_B0015)</a> <b>(Figure 1)</b>.
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In presence of PCBs, when compound diffuses into the media, it binds to bphr2 protein which undergoes a conformational change that permits to activate more stronger bphR1 promoter and increase the transcription of RFP.  
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<div class="center">
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<div class="thumb tnone">
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  <div class="thumbinner" style="width:502px;">
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  <a href="https://static.igem.org/mediawiki/parts/5/59/Evry2014_Plasmid_bphr2.jpg" class="image">
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    <img alt="IMAGE" src="https://static.igem.org/mediawiki/parts/5/59/Evry2014_Plasmid_bphr2.jpg" width="202px;" class="thumbimage"/>
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  </a>
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    <a href="https://static.igem.org/mediawiki/parts/5/59/Evry2014_Plasmid_bphr2.jpg" class="internal" title="Enlarge">
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      <img src="/wiki/skins/common/images/magnify-clip.png" width="15" height="11" alt="Symbol"/>
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    <center><b><u>Figure 1: Scheme of BBa_K1413023 construction.</u></b></center>
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<br/><u>Mechanism explanation: </u>In absence of PCBs, bphR2 <a href="https://parts.igem.org/Part:BBa_K1413021">(BBa_K1413021)</a> is bound to bphR1 promoter which activate the transcription of RFP but in very low expression.  
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In presence of PCBs, when compound diffuses into the media, it binds to bphr2 protein which undergoes a conformational change that permits to activate more stronger bphR1 promoter and increase the transcription of RFP <b>(Figure 2)</b>.  
 
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     <center><u><b>Mechanism of PCB biosensor with the system bphR2/bphR1 gene</u></b></center>
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     <center><b><u>Figure 2: Mechanism of PCB biosensor with the system bphR2/bphR1 gene</u></b></center>
 
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<!-- Add more about the biology of this part here
 
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===Usage and Biology===
 
  
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===Usage and Biology===
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<html>Combinated with the part <a href="https://parts.igem.org/Part:BBa_K1413024">BBa_K1413024</a>, it could be used like PCB biosensor.
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1413023 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1413023 SequenceAndFeatures</partinfo>

Latest revision as of 19:17, 2 November 2014

Promoter-RBS-bphr2 mutated-Terminator

bphR2 gene, from Pseudomonas pseudoalcaligens KF707, allows the transcription of bphR1 which activates the genes of degradation of biphenyls. In our project, genes of degradation was replaced by RFP gene to detect the compound. This part is composed by constitutive promoter (BBa_J23114), RBS (BBa_B0034), bphr2 (BBa_K1413021) and terminator (BBa_B0015) (Figure 1).

IMAGE
Symbol
Figure 1: Scheme of BBa_K1413023 construction.

Mechanism explanation: In absence of PCBs, bphR2 (BBa_K1413021) is bound to bphR1 promoter which activate the transcription of RFP but in very low expression. In presence of PCBs, when compound diffuses into the media, it binds to bphr2 protein which undergoes a conformational change that permits to activate more stronger bphR1 promoter and increase the transcription of RFP (Figure 2).

IMAGE
Symbol
Figure 2: Mechanism of PCB biosensor with the system bphR2/bphR1 gene

Usage and Biology

Combinated with the part BBa_K1413024, it could be used like PCB biosensor.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 568
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 352