Part:BBa_K2610020
pClpB-GFP
This reporter part features the promoter of the ClpB gene and the fluorescent reporter protein GFP (BBa_E0040). The ClpB protein is part of the stress-induced multi-chaperone system, helping E. coli cells recover from heat-induced DNA damage. (Eriksson et al, 2000)
For more information please refer to our [http://2018.igem.org/Team:Leiden/Results Results page]
Usage
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 ClpB transcription.
As can be seen in Figure 1, treatment with ascorbic acid leads to an increased fluorescence as compared to the control for low to intermediate concentrations.
Figure 1. Median Fluorescence Intensity (MFI) in AU after 4 hour incubation with ascorbic acid in low (2.5 ug/mL), intermediate (5 ug/mL) and high (10 ug/mL) concentrations. This experiment was pH controlled with hydrochloric acid.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 131
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 945
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