Tokyo/Works/Assay2

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==Activity check on lambda cI-regulated promoter and the lux lac hybrid promoter ==
==Activity check on lambda cI-regulated promoter and the lux lac hybrid promoter ==
==== Purpose: ====  
==== Purpose: ====  
-
To measure and compare the activities of two different promoters on two different types of plasmids by fluorescence of GFP downstream of each promoter
+
To measure and compare the activities of two different ''promoters + plasmid sets'' by fluorescence of GFP downstream of each promoter
 +
 
==== Samples: ====
==== Samples: ====
<br>・A4Δp pc1-GFP
<br>・A4Δp pc1-GFP

Revision as of 16:56, 24 October 2007


Works top  0.Hybrid promoter  1.Formulation  2.Assay1  3.Simulation  4.Assay2  5.Future works

Parameters for the equations in Formulation <←link> have been experimentally determined in Assay1 <←link>. Analysing the result, the following experiments were turned out to be necessary.

Activation check by cell-produced AHL

Purpose

To check if worker E. coli (Sender) can produce enough AHL for our model to work by using different copy numbers of plasmids.


Result & Conclusion

Not only high copy number plasmid pSB1, also low copy number plasmid pSB4 and pBR produced enough AHL to activate the LacI hybrid promoter in other cells. Especially, pBR remarkably produced AHL in the present experiment.
AHL assay endogenous Results.jpg

⇒see more details

Activity check on lambda cI-regulated promoter and the lux lac hybrid promoter

Purpose:

To measure and compare the activities of two different promoters + plasmid sets by fluorescence of GFP downstream of each promoter

Samples:


・A4Δp pc1-GFP
・A4 hybrid GFP PBR322TetR (+)AHL
・A4 hybrid GFP PBR322TetR (-)AHL

Procedure:

Result & Conclusion: