Difference between revisions of "Part:BBa K1163102"
 
(39 intermediate revisions by 2 users not shown)
Line 1: Line 1:
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K1163102 short</partinfo>
 
<partinfo>BBa_K1163102 short</partinfo>
−
 
+
<br>
 
<b>iGEM Evry 2013</b>
 
<b>iGEM Evry 2013</b>
  
 
<p>
 
<p>
−
Description:<br>
+
<b>Description:</b>
 +
<br>
 
This part is composed of the following elements:
 
This part is composed of the following elements:
 
<ul>
 
<ul>
−
   <li>EntA : </li>
+
   <li>pAceB promoter region</li>
−
   <li>EntB : </li>
+
   <li>superfolder GFP</li>
−
   <li>EntC : </li>
+
   <li>Terminator</li>
−
  <li>EntD : </li>
+
−
  <li>EntE : </li>
+
−
  <li>EntF : </li>
+
 
</ul>
 
</ul>
  
−
Promoter from E. coli that controls the AceB gene, involved in iron uptake.  
+
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>
 +
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>
 +
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>
 
</p>
−
This promoter sequence contains a RBS and a FUR binding site, although it is not clearly possible to identify them precisely. It has been shown to downregulate the expression of sfGFP or any gene down-stream in the presence of iron in the range: 10<sup>-7</sup> to 10<sup>-4</sup> mol.L<sup>-1</sup>
 
  
−
FUR (Ferric Uptake Regulation) is a transcriptional repressor of genes involved in iron homeostasis. In presence of iron FUR will be linked to the iron. This modification of conformation will induce a dimerization of FUR. Then the dimeric FUR will bind to the DNA in a Fur Binding Site and inhibit the mRNA transcription.
+
<br>
  
−
To caracterize our constructions we use this Fur binding site merged with a super fold GFP to the power of inhibition of the Fur binding site.
+
<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>
 +
 
 +
<br>
 +
 
 +
<p>
 +
<b>Characterization:</b>
 +
<br>
 +
We characterized its inhibition ability by cloning a superfolder GFP downstream at a iron concentration range from 10<sup>-7</sup> to 10<sup>-4</sup> mol.L<sup>-1</sup>.
 +
</p>
 +
 
 +
<br>
 +
 
 +
<p>
 +
<b>Compatibility:</b>
 +
<br>
 +
[https://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=cell ''Chassis:''] Device has been shown to work in ''<partinfo>BBa_V1004</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>''
 +
</p>
 +
 
 +
<br>
 +
 
 +
<b>Data:</b>
 +
<br>
 +
 
 +
<html>
 +
<div align="center">
 +
<img src="https://static.igem.org/mediawiki/2013/9/97/D.O._senseur.png" width="70%"/>
 +
</div>
 +
</html>
 +
 
 +
<p>
 +
<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.
 +
</p>
 +
 
 +
<br>
 +
<br>
 +
<br>
 +
 
 +
<html>
 +
<div align='center'>
 +
<img src="https://static.igem.org/mediawiki/2013/b/bf/Senseur_GFP.png" width="100%"/>
 +
</div>
 +
</html>
 +
 
 +
<p>
 +
<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.
 +
</p>
  
−
Why would you use this part?
 
  
−
This part allows the characterization of the AceB promoter region thanks to the sfGFP downstream.
+
<br>
 +
<br>
 +
<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