Device

Part:BBa_K300010

Designed by: Lorenzo Pasotti, Susanna Zucca, Nicolo' Politi   Group: iGEM10_UNIPV-Pavia   (2010-10-13)

PoPS-based self-inducible device

This is a PoPS-in/PoPS-out device.

The luxR gene (BBa_C0062) is constitutively produced by the BBa_R0040 promoter and the LuxR protein can activate the lux pR in presence of the autoinducer 3-oxo-C6-homoserine-lactone (3OC6HSL or simply HSL). The PoPS input regulates the production of luxI gene (BBa_C0061). It encodes for the LuxI enzyme, which is able to produce HSL. HSL is a very small molecule that can diffuse in the growth medium of the cells that express LuxI. The lux pR produces a PoPS output when HSL reaches a critical concentration.

This device can be specialized by assembling a promoter upstream and a promoterless expression system with the gene of interest downstream. When a cell population expresses luxI, the concentration of HSL is an increasing function of cell culture density and so the induction of the lux pR promoter occurs only when a critical cell density is reached. In this way, the production of the target protein is autoinduced by the culture itself depending on the HSL synthesis rate. The HSL synthesis rate can be tuned by assembling promoters of different strengths upstream of luxI.

This enables the construction of a library of self-inducible devices capable of starting the target protein production at a predictable culture density.


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 658
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1813


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Categories
Parameters
None