Part:BBa_K2610019
pDnaK-GFP
This reporter part features the promoter of the DnaK gene and the fluorescent reporter protein GFP (BBa_E0040).
DnaK is a major heat shock protein in Escherichia coli. It has been reported as an essential protein for cell growth at heightened temperatures. Other research has also demonstrated a role for DnaK in hydrogen peroxide resistance and reductive division.
Promoter DnaK
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 DnaK transcription (Figure 1)
Figure 2. Median Fluorescence Intensity (MFI) in AU after 4 hour incubation with antimicrobial agents in various concentrations.
We have, on request of iGEM Bordeaux, also tested hydroxymethylfurfural or HMF for stressful effects on Escherichia coli. Our data suggests a downregulation of DnaK in response to HMF, as summarised by Figure 2.
Figure 2. Median Fluorescence Intensity measured by flow cytometry after treatment with HMF. HMF in these concentrations were not lethal to bacterial cells, data not shown.
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 1110
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