Difference between revisions of "Part:BBa K1163102"
 
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This part is composed of the following elements:
 
This part is composed of the following elements:
 
<ul>
 
<ul>
−
   <li>AceB promoter region</li>
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   <li>pAceB promoter region</li>
 
   <li>superfolder GFP</li>
 
   <li>superfolder GFP</li>
 
   <li>Terminator</li>
 
   <li>Terminator</li>
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The AceB gene encodes the malate synthase A enzyme, which is involved in carbon source management. Its promoter region has been extracted from the genome of Escherichia coli. We found after genomic research that the gene is under the regulation of fur, thus meaning it is sensitive to iron concentration variations.<br>
 
The AceB gene encodes the malate synthase A enzyme, which is involved in carbon source management. Its promoter region has been extracted from the genome of Escherichia coli. We found after genomic research that the gene is under the regulation of fur, thus meaning it is sensitive to iron concentration variations.<br>
 
<br>
 
<br>
−
FUR (Ferric Uptake Regulation) is a <b>transcriptional repressor of genes involved in iron homeostasis</b>. In the presence of iron, FUR binds iron and dimerizes. This modification of conformation allows the linkage to a Fur Binding Site and inhibits the mRNA transcription of the downstream gene.
+
FUR (Ferric Uptake Regulation) is a transcriptional repressor of genes involved in iron homeostasis. In the presence of iron, FUR binds iron and dimerizes. This modification of conformation allows the linkage to a Fur Binding Site and inhibits the mRNA transcription of the downstream gene.
 
<br>
 
<br>
 
This 300 bp long promoter sequence contains a RBS and a FUR binding site, although it has not been clearly possible to identify them.
 
This 300 bp long promoter sequence contains a RBS and a FUR binding site, although it has not been clearly possible to identify them.
 +
</p>
 +
 +
<br>
 +
 +
<p>
 +
<b>Why would you use this part?</b>
 +
<br>
 +
This part is useful for iron sensing purposes in Escherichia coli. <b>The pAceB promoter represses the downstream gene.</b>
 
</p>
 
</p>
  
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<p>
 
<p>
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<b>Compatibility</b>
+
<b>Compatibility:</b>
 
<br>
 
<br>
 
[https://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=cell ''Chassis:''] Device has been shown to work in ''<partinfo>BBa_V1004</partinfo>''<br>
 
[https://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=cell ''Chassis:''] Device has been shown to work in ''<partinfo>BBa_V1004</partinfo>''<br>
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[[Plasmid backbones|''Plasmids:'']] Device has been shown to work on ''<partinfo>pSB3A3</partinfo>''<br>
+
[[Plasmid backbones|''Plasmids:'']] Device has been shown to work on ''<partinfo>pSB1A3</partinfo>''<br>
 
[[Part Types|''Devices:'']] Device has been shown to work with ''<partinfo>BBa_K1163102</partinfo>'', ''<partinfo>BBa_K1163103</partinfo>''
 
[[Part Types|''Devices:'']] Device has been shown to work with ''<partinfo>BBa_K1163102</partinfo>'', ''<partinfo>BBa_K1163103</partinfo>''
 
</p>
 
</p>
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 +
<br>
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 +
<b>Data:</b>
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<br>
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<html>
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<div align="center">
 +
<img src="https://static.igem.org/mediawiki/2013/9/97/D.O._senseur.png" width="70%"/>
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</div>
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</html>
  
 
<p>
 
<p>
−
<b>Why would you use this part?</b>
+
<b>Figure 1</b> Growth of <i>E. coli</i> strains containing the <b>A</b> the pAceB-GFP iron sensor with the iron supplementations of either 0.1, 1, 10 uM.
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</p>
 +
 
 +
<br>
 +
<br>
 
<br>
 
<br>
  
 +
<html>
 +
<div align='center'>
 +
<img src="https://static.igem.org/mediawiki/2013/b/bf/Senseur_GFP.png" width="100%"/>
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</div>
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</html>
  
 +
<p>
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<b>Fig 2</b> GFP expression of the pAceB-GFP Biobrick (BBa_K1163102) is repressed at higher iron concentrations. This construct thus functions an an iron-responsive biosensor to repress expression of the reporter gene GFP.
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</p>
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 +
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<br>
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<br>
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<br>
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 01:02, 29 October 2013

pAceB-sfGFP-Term
iGEM Evry 2013

Description:
This part is composed of the following elements:

  • pAceB promoter region
  • superfolder GFP
  • Terminator

The AceB gene encodes the malate synthase A enzyme, which is involved in carbon source management. Its promoter region has been extracted from the genome of Escherichia coli. We found after genomic research that the gene is under the regulation of fur, thus meaning it is sensitive to iron concentration variations.

FUR (Ferric Uptake Regulation) is a transcriptional repressor of genes involved in iron homeostasis. In the presence of iron, FUR binds iron and dimerizes. This modification of conformation allows the linkage to a Fur Binding Site and inhibits the mRNA transcription of the downstream gene.
This 300 bp long promoter sequence contains a RBS and a FUR binding site, although it has not been clearly possible to identify them.


Why would you use this part?
This part is useful for iron sensing purposes in Escherichia coli. The pAceB promoter represses the downstream gene.


Characterization:
We characterized its inhibition ability by cloning a superfolder GFP downstream at a iron concentration range from 10-7 to 10-4 mol.L-1.


Compatibility:
Chassis: Device has been shown to work in BBa_V1004
Plasmids: Device has been shown to work on pSB1A3
Devices: Device has been shown to work with BBa_K1163102, BBa_K1163103


Data:

Figure 1 Growth of E. coli strains containing the A the pAceB-GFP iron sensor with the iron supplementations of either 0.1, 1, 10 uM.




Fig 2 GFP expression of the pAceB-GFP Biobrick (BBa_K1163102) is repressed at higher iron concentrations. This construct thus functions an an iron-responsive biosensor to repress expression of the reporter gene GFP.





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 277
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 319