Construction and Testing

(Difference between revisions)

Revision as of 08:57, 26 October 2007

Construction Chart

Making use of flurorescent proteins(GFP and RFP) as reporter genes - use of indicuble promoters to...

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

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 as above
	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