Device

Part:BBa_K2205022

Designed by: Bradley Brown   Group: iGEM17_Newcastle   (2017-10-04)


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

Description

The part BBa_K2205022 was designed as a detector unit of the Sensynova multicellular framework for biosensors development (Figure 1) using as a template the partBBa_J33201(design by the Edinburgh team 2006) and introducing the connector 2 CS to allow the entire Sensynova framework to work.

Framework generic.jpg


Framework Yellow Variant Pic.jpg

Characterisation

A preliminary qualitative assay was carried out as an initial test for this construct. Co-cultures of Arsenic detector, processor unit (BBa_K2205012) 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 several concentrations of Arsenic (Oppb, 10ppb, 50ppb, 100ppb) to induce the expression of quorum sensing molecules and eventually achieve the reporter visualisation (Figures 3).

Ars pellets blue2.jpg

Figure 3: Pellets collected after overnight co-cultures of Arsenic detector + processor + sfGFP reporter in ratio 1:1:13, in presence of different concentrations of Arsenic.


This qualitative assay above shows that there is no significant difference among the samples when inoculated with Arsenic in different concentrations and the controls (no Arsenic). Optimisation of the Arsenic detection into the Sensynova framework is required.

References

Brenner K, Karing D, Weiss R, Arnold F (2007) Engineered bidirectional communication mediates a consensus in a microbial biofilm consortium. Proc Natl Acad Sci USA 104(44): 17300 - 17304

de Mora K, Joshi N, Balint BL, Ward FB, Elfick A, French CE (2011) A pH-based biosensor for detection of arsenic in drinking water. Anal Bioanal Chem 400(4):1031-9 (Epub 2011 Mar 27).


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 255
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 767
  • 1000
    COMPATIBLE WITH RFC[1000]


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