IGEM:IMPERIAL/2007/Experimental Design/Phase1/Results 1.2: Difference between revisions
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==Aims== | ==Aims== | ||
To Determine if the following constructs work in vivo: | |||
*[http://parts.mit.edu/registry/index.php/Part:BBa_I13522 '''pTet-GFP'''] | |||
*[http://parts.mit.edu/registry/index.php/Part:BBa_T9002 pTet-LuxR-pLux-GFP] | |||
This experiment was done to test for the viability of the construct pTet-LuxR-pLux-GFP for the applcation Infector Detector. If the construct is working, we can proceed on to characterize it using the In Vitro system. | This experiment was done to test for the viability of the construct pTet-LuxR-pLux-GFP for the applcation Infector Detector. If the construct is working, we can proceed on to characterize it using the In Vitro system. | ||
<br><br> | <br><br> | ||
Revision as of 06:38, 14 September 2007
In Vivo Testing of Constructs for Infector Detector
Aims
To Determine if the following constructs work in vivo:
This experiment was done to test for the viability of the construct pTet-LuxR-pLux-GFP for the applcation Infector Detector. If the construct is working, we can proceed on to characterize it using the In Vitro system.
Results
pTet-LuxR-pLux-GFP
The contruct was successfully tested by flooding it with a concentration of ??? nM of AHL. Since we just wanted to simply test whether the construct works or not, we did not want AHL to be a constraint and that is why a large concentration was used. Three repeated measurements were performed on three different samples and compared to a diluted concentration of GFP used as a positive control.
On the graph on the right you can see the fluorescence reading of the pLux assembly 7 hours after it has been induced with AHL. The readings of pLux are comparable to the diluted GFP solution and leads us to conclude that GFP has been expressed.
Discussion
The construct was found to work in vivo. Hence we will now go into its characterization, as well as to look at its alternative construct for the application, which is pLux-GFP. Further tests will need to be conducted to reveal its sensitivity to AHL.
