iGEM Evry 2013
This part is composed of the following elements:
- pAceB promoter region
- superfolder GFP
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.
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.
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
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21Illegal BamHI site found at 277
- 23COMPATIBLE WITH RFC
- 25COMPATIBLE WITH RFC
- 1000Illegal SapI.rc site found at 319