Difference between revisions of "Part:BBa K546000"

Revision as of 01:49, 22 September 2011

Lux pL controlled LuxR with lux pR autoinducing LuxI(lva tag)- AHL.

This device has two protein generators: lux pl-RBS-C0062-B0015 produces LuxR. LuxR forms a complex with the quorum sensing molecule AHL and this complex subsequently increases the transcriptional rate of the lux pR promoter while also decreasing the transcriptional rate of the lux pL promoter.

The quorum sensing molecule AHL is produced by LuxI which is under the control of the lux pR promoter. In combination with LuxR, AHL thus increases the production of AHL, a positive feedback loop.

Experimental data

For determining the experimental data this part was combined with K546002, together K546005. Fluorescence of a culture with OD600 of 0.6 was monitored over time in a plate reader:

The GFP expression rapidly increases, indicating functionality of this part. The formed AHL from LuxI forms a complex with LuxR and induces pR, that controls GFP expression. As a result, a positive feedback loop is initiated.

Safety Aspects

There's a small note regarding the biosafety of this BioBrick composite part. Upon release into the environment (this includes infection of a laboratory worker) this BioBrick composite part might alter the pathogenicity of pathogens that rely on quorum sensing to infect hosts. Pathogenic species that might be involved can be found on the following external [http://2011.igem.org/Team:Wageningen_UR/Safety/One#Risk_Identification_of_BioBrick_System_Inside_the_Cell_Chassis biological safety page].

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 1821
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 1101

@@ Line 12: / Line 12: @@
 ===Experimental data===
-This part was fused with GFP under control of K546003, together [https://parts.igem.org/Part:BBa_K546005 K546005]. Fluorescence of a culture with OD600 of 0.6 was monitored over time in a plate reader:
+For determining the experimental data this part was combined with [https://parts.igem.org/Part:BBa_K546002 K546002], together [https://parts.igem.org/Part:BBa_K546005 K546005]. Fluorescence of a culture with OD600 of 0.6 was monitored over time in a plate reader:
 [[Image: Fluorescence measurement BBa K546005-BBa K546001.png]]
 The GFP expression rapidly increases, indicating functionality of this part. The formed AHL from LuxI forms a complex with LuxR and induces pR, that controls GFP expression. As a result, a positive feedback loop is initiated.
 ===Safety Aspects===