From 2007.igem.org
Construction
Testing/Results
Testing Constructs
Making use of flurorescent proteins(GFP and RFP) as reporter genes
- use of indicuble promoters to...
Comparing basal and induced expression levels for the quorum sensing promoters
| CONSTRUCT |
AIMS |
 |
The quorum-sensing promoters: How active is 'unactivated' Plas and Plux?
- preliminary test for the basal levels of activity of the positively-controlled quorum sensing promoters
|
 |
The quorum-sensing constructs: How off is 'off' and how on is 'on'?</br>
- test for levels of expression in the unenhanced versus enhanced state
- xR must be expressed in same strain as Px; xI can be expressed in a different strain since its AHL is diffusable
- both strains grown in a single culture tube
|
Testing functionality of the various promoters
| CONSTRUCT |
AIMS |
 |
The Omp promoter
- postively-controlled promoter activated by endogenous OmpR protein (sensitive to salt concentrations via EnvZ protein)
- test for activity using high salt levels in EnvZ+ strain
- test for activity using the photo-active red system in EnvZ- strain
|
 |
The Tet promoter and operator
- negatively-controlled promoter, repressed by TetR and derepressed by the inducer tetracycline
- test for levels of expression during repressed versus derepressed state
|
 |
The quorum-sensing promoters: Plas and Plux
|
 |
The PlasOcI promoter-operator fusion
- negatively- and positively-controlled promoter in the final half adder design, repressed by cI and enhanced by LasR
- testfor activity in the repressed versus enhanced state
- test in addition to above construct since operator fusion may affect promoter activity
|
