Signalling

Part:BBa_K3267000

Designed by: Yuma Kanazawa   Group: iGEM19_NYU_Shanghai   (2019-10-15)
Revision as of 22:28, 21 October 2019 by Yumakk (Talk | contribs) (→‎Usage)


Bioelectric Relay Construct

This part will respond to the electric potential via various redox modulators and the transformed cell begins to have the activated transcription factor, which is SoxR protein. This transcription factor protein binds to the bidirectional promoter region, then cells translate more SoxR and LuxI.

Biology

The Relay part contains E.coli. intrinsic the bidirectional promoter region of the SoxRS regulon. The SoxR coding sequence is truncated and LuxI coding region is added followed by a ssRA tag. LuxI gene is originally from the bacterium A. fischeri and encodes Acyl-homoserine-lactose synthase. LuxI synthases Acyl-homoserine-lactose (AHL) which is known as autoinducing peptides. The peptides are released in the extracellular environment, and other cells are able to detect the change in concentration of this peptide. This autoinduction process is called quorum-sensing (QS).

Usage

In our device, we use multiple redox modulators to convert electric stimuli into chemical signals. [Fig 1] The toxin of Gram-negative bacterium Pseudomonas aeruginosa, Pyocyanin is used as the initiator of this redox cycle. Added pyocyanin is the oxidized form and it oxidizes SoxR Transcription Factor protein to activate. The ubiquitous Menaquinone transports an electron across the membrane and Pyocyanin is again oxidized. Ferrocyanide/Ferricyanide in the extracellular environment gets reduced via Menaquinone and the positive potential oxidizes Ferrocyanide to keep modulators oxidized. Therefore, the cycle keeps activating SoxR protein. The activated SoxR protein binds to the pSoxR/pSoxS bidirectional promoter region and expresses LuxI and SoxR proteins. SoxR protein is again activated under the positive electric potential as explained above. LuxI protein acts as Acyl-homoserine lactone (AHL) synthase. AHL will be diffused outside the cell and induces the surrounding cells' response to the change in the concentration of AHL.

Characterization

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1165
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 49


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