Difference between revisions of "Part:BBa K1163102"
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<b>Compatibility</b>
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[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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Figure 1 - Transfer function of <partinfo>F2620</partinfo>.  The data points represent the mean of 6 or 9 individual measurements.  The corresponding error bars represent the 95% confidence interval in the mean of the independent measurements. The blue shaded region represents the range bounded by the lowest and highest outputs among the independent measurements at a given input level.  The solid black curve was calculated by fitting a simple Hill function to the experimental measurements.
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<b>Figure 1</b> Growth of <i>E. coli</i> strains containing either the <b>A</b> the pAceB-GFP iron sensor or the <b>B</b> pAceB-lacI+PL_lacO-RFP Fur inverter with the iron supplementations of either 0.1, 1, 10, 100 uM.
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<img src="https://static.igem.org/mediawiki/2013/6/6a/Fig3_aceB-GFP.jpg" alt="pAceB_GFP_characterization" width="70%"/>
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<b>Fig 2</b> GFP expression of the pAceB-GFP Biobrick <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K1163102:Experience">BBa_K1163102</a> 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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<img src="https://static.igem.org/mediawiki/2013/f/f2/LacO_RFP.jpg" alt="OD_pAceB_GFP" width="70%"/>
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<b>Fig 3</b> RFP expression of <i>E. coli</i> transformed with plLacO-RFP (Biobrick BBa_J04450) in presence (plates 1 and 2) and absence (plate 3) of IPTG. Adding IPTG increases RFP expression, confirming that Biobrick BBa_J04450 enables LacI-mediated RFP repression.</p>
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 01:16, 5 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:
OD_pAceB_GFP


<p> Figure 1 Growth of E. coli strains containing either the A the pAceB-GFP iron sensor or the B pAceB-lacI+PL_lacO-RFP Fur inverter with the iron supplementations of either 0.1, 1, 10, 100 uM.

pAceB_GFP_characterization




Fig 2 GFP expression of the pAceB-GFP Biobrick <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K1163102:Experience">BBa_K1163102</a> is repressed at higher iron concentrations. This construct thus functions an an iron-responsive biosensor to repress expression of the reporter gene GFP.

OD_pAceB_GFP




Fig 3 RFP expression of E. coli transformed with plLacO-RFP (Biobrick BBa_J04450) in presence (plates 1 and 2) and absence (plate 3) of IPTG. Adding IPTG increases RFP expression, confirming that Biobrick BBa_J04450 enables LacI-mediated RFP repression.


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