Difference between revisions of "Part:BBa K3484004"
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+ | == Model Characterization == | ||
+ | From the ODE system (Eq.1) a transfer function can be derived if LuxR and GFP are on a steady state (dy/dx=0). This transfer function (Eq.2) allows us to connect the concentration of lactone (AHL) in the media to the fluorescence that the sensor cells are emitting. The function has the shape of a Hill function with a low hill coefficient (n=1), thus meaning that the sensor will not be hypersensitive and it will have a long dynamic range where it can sense the AHL concentration[1][2]. | ||
+ | |||
+ | [[File:UPF_Barcelona--Reporter3.png|600px|thumb|center|Equation 2. Transfer function of the AHL sfGFP+ASV biosensor model]] | ||
+ | |||
+ | [1] Ballestero, M. C., Duran-Nebreda, S., Monta, R., Solé, R., Macía, J., Rodríguez-Caso, C. A bottom-up characterization of transfer functions for synthetic biology designs: lessons from enzymology. Nucleic Acids Research, 2014, Vol. 42, No. 22. | ||
+ | |||
+ | [2] Garcia-Ojalvo, J., Elowitz, M.B., Strogatz, S.H. Modeling a synthetic multicellular clock: repressilators coupled by quorum sensing. Proc. Natl Acad. Sci. U.S.A., 101, 10955–10960. | ||
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Revision as of 12:07, 26 October 2020
AHL sfGFP+ASV reporter
This composite part produces sfGFP when a large enough concentration of lactone (AHL) is present in the medium. In this case, it constitutively produces LuxR, which binds to the lactone present in the medium and together, they form a complex that activates the pLux promoter, generating the sfGFP whose fluorescence can be measured. The schematics of this composite part can be seen in Figure 1.
This system allows a fast report on the AHL concentrations in the medium, something important to know the amount of protein. The composite part is Biobrick compatible.
This composite part is composed by all the following parts:
BBa_B0015: A reliable double terminator that consists on BBa_B0010 and BBa_B0012.
BBa_B0033: A weak RBS. The RBS.4 (derivative of BBa_0030).
BBa_B0032: A medium RBS. The RBS.3 (medium) (derivative of BBa_0030
BBa_R0062: LuxR & HSL regulated promoter.
BBa_J72005: Ptet Promoter. Is activated by the AHL-LuxR complex
BBa_K3484006: sfGFP+ASV coding sequence
Model Characterization
From the ODE system (Eq.1) a transfer function can be derived if LuxR and GFP are on a steady state (dy/dx=0). This transfer function (Eq.2) allows us to connect the concentration of lactone (AHL) in the media to the fluorescence that the sensor cells are emitting. The function has the shape of a Hill function with a low hill coefficient (n=1), thus meaning that the sensor will not be hypersensitive and it will have a long dynamic range where it can sense the AHL concentration[1][2].
[1] Ballestero, M. C., Duran-Nebreda, S., Monta, R., Solé, R., Macía, J., Rodríguez-Caso, C. A bottom-up characterization of transfer functions for synthetic biology designs: lessons from enzymology. Nucleic Acids Research, 2014, Vol. 42, No. 22.
[2] Garcia-Ojalvo, J., Elowitz, M.B., Strogatz, S.H. Modeling a synthetic multicellular clock: repressilators coupled by quorum sensing. Proc. Natl Acad. Sci. U.S.A., 101, 10955–10960.
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 1249
Illegal AgeI site found at 1372 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1142