Difference between revisions of "Part:BBa K300010"

Revision as of 19:44, 27 October 2010

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 it 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. The produced HSL can diffuse in the growth media 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 the cells reach a threshold density.

In this way, the upstream promoter autoinduces the production of the target protein at a critical culture density, 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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 -
+This is a PoPS-in/PoPS-out device.
+The luxR gene (<partinfo>BBa_C0062</partinfo>) is constitutively produced by the <partinfo>BBa_R0040</partinfo> promoter and it 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 (<partinfo>BBa_C0061</partinfo>). It encodes for the LuxI enzyme, which is able to produce HSL. The produced HSL can diffuse in the growth media 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 the cells reach a threshold density.
+In this way, the upstream promoter autoinduces the production of the target protein at a critical culture density, 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.