Part:BBa_K916001

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-NGFP translational Fusion

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

This is for use with part BBa_K916002 as a [http://en.wikipedia.org/wiki/Bimolecular_fluorescence_complementation GFP-fragment complementation] based biosensor.

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

Usage and Biology

This is a fusion protein consisting of [http://www.ncbi.nlm.nih.gov/gene/1224220 TraR] from [http://en.wikipedia.org/wiki/Agrobacterium_tumefaciens Agrobacterium tumefaciens] and the N-terminal fragment of GFP (NGFP).

TraR is the A. tumefaciens transcriptional regulator responsible for its [http://en.wikipedia.org/wiki/Quorum_sensing quorum sensing] response. It binds N-(3-Oxo-octanoyl)-l-homoserine lactone (3-oxo-C8-HSL), and upon binding forms a homodimer. Most similar [http://en.wikipedia.org/wiki/LuxR-type_DNA-binding_HTH_domain 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
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 6
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1233
  Illegal XhoI site found at 521
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]