Coding

Part:BBa_K916002

Designed by: Joseph Elsherbini   Group: iGEM12_Georgia_Tech   (2012-10-01)


This part is a work in progress and does not yet work as expected.

See a full description of the progress [http://2012.igem.org/Team:Georgia_Tech/Project on our wiki].

TraR-CGFP Translational Fusion

NOTE: This part has no promoter, and so can be used with any BioBrick promoter in the registry. See the list of promoters here.

Figure 1 Barnard E, McFerran NV, Trudgett A, Nelson J, Timson DJ. Development and implementation of split-GFP-based bimolecular fluorescence complementation (BiFC) assays in yeast. Biochem Soc Trans. 2008 Jun;36(Pt 3):479-82. PMID: 18481985.
http://www.ncbi.nlm.nih.gov/pubmed/18481985
3-oxo-C8-HSL Figure 1 Luo ZQ, Su S, Farrand SK. In situ activation of the quorum-sensing transcription factor TraR by cognate and noncognate acyl-homoserine lactone ligands: kinetics and consequences. J Bacteriol. 2003 Oct;185(19):5665-72. PMID: 13129937 http://www.ncbi.nlm.nih.gov/pubmed/13129937

Usage and Biology

This is a fusion protein consisting of [http://www.wikigenes.org/e/gene/e/1224220.html TraR] from [http://en.wikipedia.org/wiki/Agrobacterium_tumefaciens Agrobacterium tumefaciens] and the C-terminal fragment of GFP (CGFP). This is for use with part BBa_K916001 as a [http://en.wikipedia.org/wiki/Bimolecular_fluorescence_complementation GFP-fragment complementation] based biosensor.

TraR is the A. tumefaciens transcriptional regulator responsible for its quorum sensing response. It binds N-(3-Oxo-octanoyl)-l-homoserine lactone (3-oxo-C8-HSL), and upon binding forms a homodimer. Most similar LuxR family quorum sensing regulators dimerize without their cognate autoinducer, but TraR will only dimerize upon binding 3-oxo-C8-HSL. We exploit this property of TraR by fusing it to two seperate fragments of GFP (NGFP and CGFP). Our expectation is that in the presence of its autoinducer, TraR will dimerize and bring the two fragments of GFP into proximity, allowing them to interact and fluoresce like an intact GFP peptide.

Please see the design notes and [http://2012.igem.org/Team:Georgia_Tech/Project our wiki] for additional information.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 984
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 910


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