Part:BBa_K2610031
pSoxS-GFP
This composite part features the regulatory part promoter pSoxS (BBa_K2610030) and the fluorescent protein GFP (BBa_E0040). It can be used to visualize upregulation of SoxS as a result of superoxide stress.
Regulatory protein SoxS is involved in the oxidative stress signaling pathway in Escherichia coli. Intracellular superoxide-generating compounds cause the SoxR to activate transcription of SoxS, which then triggers a set of defense and repair genes that form the oxidative response system.
Usage and Biology
iGEM Leiden 2018 has created a screening system for the detection of new antibiotics. This composite part which allowed us to detect stress-induced changes in SoxS transcription, signaling that a certain compound causes superoxide stress.
We have tested of these several stress-activated promoters against a repertoire of known antibiotics using flow cytometry. As can be seen in the heatmap below, treatment with both nalidixic acid and hydrogen peroxide causes a significant increase in fluorescent signal of pSoxS-GFP.
Figure 2. Heatmap with stress-activated promoters. Values for each promoter are normalised to its respective negative control (no stressor, not shown).
Figure 1. Median Fluorescence Intensity (MFI) in AU after 4 hour incubation with nalidixic acid in various concentrations.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 947
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