Part:BBa_K2587027
luxI_luxR_Plux_gfp
This construct comprises parts from the Quorum sensing system from Vibrio fischeri. It contains the LuxI, the acyl homoserine lactone synthase, the LuxR, the regulator, which will bind to the promoter pLux, that will induce synthesis of GFP.
Usage and Biology
- Acyl homoserine lactone synthase: LuxI
- Regulator: LuxR
- Promoter: pLux, inducible by the quorum sensing molecule acyl homoserine lactone bound to LuxR
- GFPfluorescent protein
- GFP expression is induced after synthesis of the acyl homerine lactone by LuxI, which binds with LuxR to the pLux promoter
- This construct shows functionality of promoter by increase of fluorescence upon time
- This part contains Type II S restriction sites and can be further used for scarless assembly with other parts
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 829
Illegal NheI site found at 852 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1082
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 892
Illegal BsaI site found at 1681
Illegal BsaI.rc site found at 1523
Illegal BsaI.rc site found at 1790
Illegal BsaI.rc site found at 2468
Characterization
Characterisation of promoter by fluorescence measurement of GFP
To check whether the promoter, pLux, is really induced by the binding of the acyl homoserine lactone to the LuxR, this construct is created. Here the relative fluorescence units are measured in comparison to the wild type E. coli cells.
Results It can clearly be observed, that fluorescence increases upon growth of the cells. Wild type cells, as a negative control, show little or no fluorescence at all.
Figure 1: Relative fluorescence measurement of lysis construct and E.coli wild type cells (negative control). Exctinction: 485nm; Emission: 520nm. Cells were measured only within a time span of 24h.
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