Device

Part:BBa_K2205023

Designed by: Lais Takiguchi   Group: iGEM17_Newcastle   (2017-10-27)


Sensynova Framework (Psicose Detector + Connector 1A (complete) - dCell)

Description

The part BBa_K2205023 was designed as a detector unit of the Sensynova multicellular framework for biosensors development (Figure 1) as a collaboration with the Evry Paris-Saclay 2017 team.

Framework generic.jpg

This part was made to test the modularity of the system by replacing the IPTG detector module of the Sensynova platform with the psicose biosensor created by the Evry Paris-Saclay 2017 team.

This part consists of the Evry psicose biosensor controlling the production of part BBa_K2205008, our connector 1.

This system works through the repression of the psicose sensitive synthetic promoter except in the presence of psicose, freeing up the promoter and subsequently activating the transcription of connector 1 and therefore triggering the following modules of the platform.


Framework Yellow Variant Pic.jpg

Characterisation

A preliminary qualitative assay was carried out as an initial test for this construct. Co-cultures of Psicose detector, processor unit and sfGFP reporter(BBa_K2205015) were inoculated and grown overnight in LB+chloramphenicol (12,5ng/ul).

The day after the cultures were diluted at OD600: 0,1 and mixed together to obtain co-cultures with ratio 1:1:13 (detector:processor:reporter). The samples were supplemented with 33.22 mM Psicose to induce the expression of quorum sensing molecules and eventually achieve the reporter visualisation (Figures 3).

Psi_pellets_gfp2.jpg

Figure 3: Pellets collected after overnight co-cultures of Psicose detector + processor + sfGFP reporter in ratio 1:1:13, with and without PSI.

References

iGEM Community. (2017). Team Evry Paris-Saclay 2017. [online] Available at: http://2017.igem.org/Team:Evry_Paris-Saclay [Accessed 30 Oct. 2017].


Sequence and Features

Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 65
    Illegal AgeI site found at 752
    Illegal AgeI site found at 1568
  • 1000
    COMPATIBLE WITH RFC[1000]


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