Part:BBa_K1886001
AHL single-cycled oscillation
Two molecules of luxR binds with AHL (catalyzed by luxI), and this compound will promote the activity of pluxR. pluxR controls the expression of gene aiiA , encoding the enzyme which degrades AHLs. And all of these form an AHL single-cycled oscillation.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1821
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 3256
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1101
Illegal BsaI.rc site found at 1970
Illegal BsaI.rc site found at 3073
Characterization
BACKGROUND
Principle
The synchronized oscillator design is based on elements of the quorum sensing machineries in Vibrio fischeri and Bacillus Thurigensis. We placed the luxI (from V. fischeri), aiiA (from B. Thurigensis) and yemGFP genes(which is used as a reporter and knocked out after the combination with logical gate) under the control of three identical copies of the luxI promoter. The LuxI synthase enzymatically produces an acyl-homoserine lactone (AHL), which is a small molecule that can diffuse across the cell membrane and mediates intercellular coupling. It binds intracellularly to the constitutively produced LuxR, and the LuxR–AHL complex is a transcriptional activator for the luxI promoter. AiiA negatively regulates the promoter by catalysing the degradation of AHL. This network architecture, whereby an activator activates its own protease or repressor, is similar to the motif used in other synthetic oscillator designs and forms the core regulatory module for many circadian clock networks. Furthermore, theoretical work has shown how the introduction of an autoinducer in similar designs can potentially lead to synchronized oscillations over a population of cells.
RESULTS
Gel electrophoretic analysis
stable oscillation in microfluidics chips
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