Difference between revisions of "Part:BBa K546005"

Revision as of 19:20, 21 September 2011

Lux pL controlled LuxR + lux pR autoinducing LuxI (lva tag) + lux pR controlled GFP(lva tag).

This BioBrick composite part has an, colony wide, initially increasing and finally constant Green Fluorescent Protein (GFP) with a LVA Tag output.

Usage and Biology

This device has three 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. Both of the other generators are under control of this lux pR promoter.

The second generator ‘luxpR-RBS-J04031-B0015’has a low basal expression of the GFP, which can - of course - be monitored.

The third generator produces luxI, which produces the quorum sensing molecule AHL. In combination with LuxR, AHL thus increases the production of AHL and GFP.

Safety Aspects

This BioBrick part produces an autoinducer that might interact with [http://2011.igem.org/Team:Wageningen_UR/Safety/One#Risk_Identification_of_BioBrick_System_Inside_the_Cell_Chassis some pathogens].

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 3909
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 1101
Illegal BsaI.rc site found at 1828
Illegal BsaI.rc site found at 2086
Illegal BsaI.rc site found at 3189

@@ Line 7: / Line 7: @@
 ===Usage and Biology===
-LuxR is slightly produced by the sub-[https://parts.igem.org/Part:BBa_K546003 BioBrick part] K546003 at the beginning. Meanwhile also the mildly transcribed BioBrick part C0061 creates an increase in the [http://en.wikipedia.org/wiki/Autoinducer autoinducer] 3OC6HSL. As the LuxR and the autoinducer are formed simultaneously, a complex of these two forms -in all the cells of a colony. This complex initiates a positive and two times a negetive feedback loop. The gene luxI is part of the positive feedback loop and will be transcribed more often (resulting in a greater autoinducer production), but the promoter in front of the luxr gene will down regulate the formation of LuxR. The increase in and finally more or less equilibrium state of the LuxR-autoinducer complex is monitored by the Green Fluorescent Protein with a LVA Tag.
+This device has three 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. Both of the other generators are under control of this lux pR promoter.
+The second generator  ‘luxpR-RBS-J04031-B0015’has a low basal expression of the GFP, which can - of course - be monitored.
+The third generator produces luxI, which produces the quorum sensing molecule AHL. In combination with LuxR, AHL thus increases the production of AHL and GFP.
 ===Safety Aspects===