Part:BBa_K2610035
pSoxS-GFP-GFP-pSoxS-GFP-GFP
This composite part features twice the regulatory part promoter SoxS (BBa_K2610030) and two times 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 superoxide pathway in Escherichia coli. It acts a superactivator of downstream stress genes.
Usage and Biology
Usage and Biology=
We, iGEM Leiden 2018, have designed this composite part as part of our project Fifty Shades of Stress. This reporter part allowed us to detect stress-induced changes in SoxS transcription. We have created this part in order to amplify the fluorescent signal measured after pSoxS-GFP activation.
As can be observed in Figure 1 the pSoxS-GFP-pSoxS-GFP construct leads to an increased mean fluorescence intensity compared to pSoxS-GFP, but not compared to pSoXS-GFP-pSoxS-GFP .
We also assessed the dose-dependency of this reporter. We found that an increase in nalidixic acid concentration of nalidixic acid leads to an increasing mean fluorescence intensity (MFI). A decrease in detected GFP expression at higher concentrations can be attributed to the lethality of the stressor.
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Figure 1. Median fluorescence intensity (MFI) as a result of treatment with nalidixic acid for pSoxS-GFP-GFP-pSoxS-GFP-GFP compared to pSoxS-GFP-pSoxS-GFP and pSoxS-GFP . It can also be seen that there is a dose response curve to nalidixic acid. The decrease in detected GFP expression at higher concentrations can be attributed to lethality of the stressor.
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
Illegal BsaI.rc site found at 1673
Illegal BsaI.rc site found at 2704
Illegal BsaI.rc site found at 3430
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