Part:BBa_K2610017
pMicF-GFP
This reporter part features the basic part promoter MicF (BBa_K2610007) and the fluorescent reporter protein GFP (BBa_E0040).
MicF small RNA, coded by the MicF gene, is produced as stress response in Escherichia coli. It negatively regulates translation of the OmpF porin. MicF binds to OmpF mRNA and represses production of OmpF in response to environmental and internal stresses. The MicF expression is stimulated by many factors such as oxidative stress, osmolarity increase and temperature increase. (Delihas et al, 2001)
More in depth information can be found on our [http://2018.igem.org/Team:Leiden/Results results page]
pMicF as a non-specific stress indicator
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 MicF transcription.
Figure 1. Median Fluorescence Intensity (MFI) in AU after 4 hour incubation with antimicrobial agents in various concentrations. Luria Bertani broth was used as a control measurement.
In order to validate our newly designed part, we performed flow cytometry after stressing our transformed DH5A cells with nalidixic acid, tetracycline, chloramphenicol and magnesium chloride in various concentrations. As can be seen in Figure 1, all of our chosen antibiotics slightly increased the MFI after 4 hours of incubations. This supports suggestions in literature that MicF is upregulated by copious environmental and internal stresses.
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 951
| None |