Team:Paris/Addressing testing

From 2009.igem.org

iGEM > Paris > WetLab > Addressing

Menu accordéon avec jQuery

WetLab - Addressing the message

Main | Addressing| Production| Reception

Global constructions :

The green color means the experiment was a success

Experiments ran :

Column 1	Column 2	Column 3	Column 4
PCR : pBAD: plate 2008 ClyA Cterm matrix : F4 Oligo :O10 and O31 TM :55°C mRFP Nterm plasmid : pSB1A3 Oligo :O57 and O58 TM : ClyA Nterm matrix : F4 Oligo : O32 and O9 TM :55°C mRFP Cterm matrix : pSB1A3 Oligo :O59 and O60 TM :	-	-	-
Verification on gel : ok	-	-	-
Purification on gel : ok	-	-	-
Digestion: ClyA Cterm S/P RFP Nterm X/P pBAD vector : ? E/S	Digestion: ClyA Cter-RFP Nter vector PSB1A3 E/X	-	-
Verification digestion: ok	Verification digestion: ok	-	-
Ligation: ClyA Cter-RFP Nter vector PSB1A3	Ligation: PBAD E/S ClyA Cter-RFP Nter PSB1A3 E/X (x2) PBAD S/P ClyA Cter-RFP Nter PSB2K3 X/P (x2)	Ligation: -
Colony PCR : ok	Colony PCR : ok	Colony PCR : ok
Miniprep: clone :	Miniprep: clone :	Miniprep: clone 3 clone 5
Sequencing : ok	Sequencing : ok	Sequencing : ok	-
Stock glycerol: ClyA CTer: S47(clone 3) and S48(clone 7) ClyA Nter: S72 RFP Cter: S55 (Clone 3) RFP Nter: S56 (Clone 3)	Stock glycerol Cly A(Cter)-(Nter)RFP: S72(clone 8)	Stock glycerol pBAD ClyA RFP : S89 (clone 3) pBAD ClyA RFP: S90 (clone 5)

Functional Testing:

PBAD ClyA RFP was transformed into Top10 bacteria in order to localize the fluorescence, we are supposed to have a superior fluorescence in the membrane.

PBAD ClyA RFP on PSB3T5 was transformed into Delta Tol bacteria , in this case we are supposed to see fluorescent vesicles when the medium contains 1 % arabinose , and to have no fluorescent on 1% glucose.

To learn more, see the Conclusion page

Export system

We finally thought that it won't be neccesary to overexpress the Tat system, nevertheless we have run a few experiments before starting to focus on others parts of the project.

Experiments ran :

Column 1	Column 2	Column 3	Column 4
PCR : TatABCE matrix :	-	-	-
Verification on gel : ok	-	-	-
Purification on gel : ok	-	-	-
Digestion: none	Digestion: none	Digestion: none	Digestion: none
Digestion checking: none	Digestion checking: none	Digestion checking: none	Digestion checking: none
Ligation: none	Ligation: none	Ligation: none	Ligation: none
Colony PCR : none	Colony PCR : none	Colony PCR : none	Colony PCR : none
Miniprep: none	Miniprep: none	Miniprep: none	Miniprep: none
Sequencing : none	Sequencing : none	Sequencing : none	Sequencing : none
Stock glycerol: none	Stock glycerol none	Stock glycerol none	Stock glycerol none