Part:BBa_K4634003
pPepT Hypoxia-Induced Reporter System
pPepT, which is induced by hypoxia, is regulated by transcriptional activator: fumarate and nitrate reduction regulatory protein (FNR).[1] Under anaerobic conditions, FNR associates with [4Fe-4S]2+ clusters to form transcriptionally active homodimers, the dimer can bind DNA site-specifically and regulate transcription from target promoters. However, when the clusters are disrupted in the presence of oxygen, the FNR dimer dissociates into nonDNA-binding monomers. [2, 3] In order to verify the anaerobic induction function of pPepT, we used pPepT and mcherry to form a reporter system. Fluorescence intensity was verified separately in normoxic and anaerobic environments. At the same time, a reporting system under the control of T7 promoter was also set up as a control group. The results show that for the pPepT experimental group, induction under anaerobic conditions can indeed increase the intensity of the reporter fluorescence.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 995
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 958
- 1000COMPATIBLE WITH RFC[1000]
References
[1]Yu, Bin, et al. "Explicit hypoxia targeting with tumor suppression by creating an “obligate” anaerobic Salmonella Typhimurium strain." Scientific reports 2.1 (2012): 436.
[2]Crack, Jason C., et al. "Signal perception by FNR: the role of the iron sulfur cluster1." (2008): 1144-1148.
[3] CRACK J C, GASKELL A A, GREEN J, et al. Influence of the Environment on the [4Fe−4S]2+ to [2Fe−2S]2+ Cluster Switch in the Transcriptional Regulator FNR [J]. Journal of the American Chemical Society, 2008, 130(5): 1749-58.
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