Difference between revisions of "Part:BBa K1413024"

 
 
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<partinfo>BBa_K1413024 short</partinfo>
 
<partinfo>BBa_K1413024 short</partinfo>
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This part is composed by bphR1 promoter <a href="https://parts.igem.org/Part:BBa_K1155001">(BBa_K1155001)</a>, from Pseudomonas pseudoalcaligenes KF707, which allows the transcription of the genes of biphenyl degradation, the RBS <a href="https://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, RFP gene <a href="https://parts.igem.org/Part:BBa_E1010">(BBa_E1010)</a> and terminator <a href="https://parts.igem.org/Part:BBa_B0015">(BBa_B0015)</a> <b>(Figure 1)</b>. In our project, genes of degradation was replaced by RFP gene to detect the compound.
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  <a href="https://static.igem.org/mediawiki/parts/f/f4/Evry2014_Plasmid_bphr1.jpg" class="image">
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    <img alt="IMAGE" src="https://static.igem.org/mediawiki/parts/f/f4/Evry2014_Plasmid_bphr1.jpg" width="202px;" class="thumbimage"/>
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    <a href="https://static.igem.org/mediawiki/parts/f/f4/Evry2014_Plasmid_bphr1.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><u><b>Figure 1: Schema of BBa_K1413024 construction.</b></u></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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  <a href="https://static.igem.org/mediawiki/2014/3/3f/Igemevry_2014_Sch%C3%A9ma_g%C3%A9n%C3%A9ral_pcb.jpg" class="image">
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    <img alt="IMAGE" src="https://static.igem.org/mediawiki/2014/3/3f/Igemevry_2014_Sch%C3%A9ma_g%C3%A9n%C3%A9ral_pcb.jpg" width="402px;" class="thumbimage"/>
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  </a>
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    <a href="https://static.igem.org/mediawiki/2014/3/3f/Igemevry_2014_Sch%C3%A9ma_g%C3%A9n%C3%A9ral_pcb.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><u><b>Figure 2 : Mechanism of PCB biosensor with the system bphR2/bphR1 gene </b></u></center>
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bphR1 promoter, from Pseudomonas pseudoalcaligenes KF707, activates the bph genes of biphenyls degradation, in presence of bphR2 protein and biphenyl.
 
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In our study, the aim was to detect the presence of polychlorinated biphenyls (PCBs) with RFP fluorescence.
 
  
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===Usage and Biology===
 
===Usage and Biology===
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<html>Combinated with the part <a href="https://parts.igem.org/Part:BBa_K1413023">BBa_K1413023</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_K1413024 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1413024 SequenceAndFeatures</partinfo>

Latest revision as of 19:21, 2 November 2014

bphR1-RBS-RFP-Terminator

This part is composed by bphR1 promoter (BBa_K1155001), from Pseudomonas pseudoalcaligenes KF707, which allows the transcription of the genes of biphenyl degradation, the RBS (BBa_B0034), RFP gene (BBa_E1010) and terminator (BBa_B0015) (Figure 1). In our project, genes of degradation was replaced by RFP gene to detect the compound.

IMAGE
Symbol
Figure 1: Schema of BBa_K1413024 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_K1413023, it could be used like PCB biosensor.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 156
    Illegal BamHI site found at 239
    Illegal XhoI site found at 46
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 883
    Illegal AgeI site found at 995
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 139