Difference between revisions of "Part:BBa K3470015"
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==Circuit== | ==Circuit== | ||
'''SoxS promoter – RBS – GFP - Double Terminator''' | '''SoxS promoter – RBS – GFP - Double Terminator''' | ||
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The GFP intensity is expected to increase with time for all the concentrations except the control and stabilize after a few hours. The culture fluorescence is hypothesized to be more for a higher concentration of paraquat and then decrease after a point due to bacterial death. Bacterial resistance to oxidative stress must be priorly checked. | The GFP intensity is expected to increase with time for all the concentrations except the control and stabilize after a few hours. The culture fluorescence is hypothesized to be more for a higher concentration of paraquat and then decrease after a point due to bacterial death. Bacterial resistance to oxidative stress must be priorly checked. | ||
| + | ==Sequence and features== | ||
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| + | <partinfo>BBa_K3470015 SequenceAndFeatures</partinfo> | ||
==References== | ==References== | ||
Latest revision as of 14:03, 23 October 2020
Anti inflammation control mechanism (without SoxR)
Contents
Circuit
SoxS promoter – RBS – GFP - Double Terminator
Usage and Biology
SoxS is a conditional promoter, regulated by the SoxR transcription factor (In Composite BioBrick 1). In the active state (when it is bound to NO; pro-inflammatory signal), SoxR activates the SoxS promoter and starts the transcription of the genes downstream to it. (Hidalgo et al., 1998).
Proposed Experimentation
Escherichia coli Nissle 1917 (bacteria) transformed with SoxR and SoxS genes must be tested to analyze the relationship between the amount of oxidative stress and transcription rates of the genes in response to methyl viologen dichloride hydrate which acts as an inducer by measuring GFP intensity. The intensity of GFP is expected to increase with time indicating that even if the concentration of the oxidative stress inducer is constant, the rate of activation of SoxR and thus the rate of transcription of SoxS promoter increases and then stabilizes to a constant rate. It is also expected that transcription rate of SoxS promoter increases with a higher inducer concentration.
It is also expected that the amount of oxidative stress required to activate SoxR and in turn activate SoxS promoter occurs even at low oxidative stress, preventing any unwanted inflammation due to our probiotic thus getting an anti-inflammatory response activation by composite bio-brick 2.
The GFP intensity is expected to increase with time for all the concentrations except the control and stabilize after a few hours. The culture fluorescence is hypothesized to be more for a higher concentration of paraquat and then decrease after a point due to bacterial death. Bacterial resistance to oxidative stress must be priorly checked.
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 732
References
Hidalgo, E., Leautaud, V., & Demple, B. (1998). The redox-regulated SoxR protein acts from a single DNA site as a repressor and an allosteric activator. EMBO Journal, 17(9), 2629–2636. https://doi.org/10.1093/emboj/17.9.2629
Remington, S. J. (2011). Green fluorescent protein: A perspective. Protein Science, 20(9), 1509–1519. https://doi.org/10.1002/pro.684