Tokyo/Works/Assay

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====Result & Conclusion====
====Result & Conclusion====
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== [[Tokyo/Activation check by cell-produced AHL |Activation check by cell-produced AHL ]] ==
 
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==== Purpose ====
 
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To check if worker E. coli (Sender) can produce enough AHL for our model to work by using different copy numbers of plasmids.
 
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====Result & Conclusion====
 
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[[Image:AHL_assay_endogenous_Results.jpg]]
 
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<br>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.
 
=2.Preliminary assays=
=2.Preliminary assays=
'''We conducted preliminary assays to practice experiments and check if the materials and methods worked properly.'''  
'''We conducted preliminary assays to practice experiments and check if the materials and methods worked properly.'''  
<br>[[Tokyo/Preliminary assays |See more details]]
<br>[[Tokyo/Preliminary assays |See more details]]

Revision as of 02:50, 24 October 2007


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

1.Important assay

Hill function fitting

新しく作ったハイブリッドプロモーターに関してはデータがないので,実験によりパラメータを求める. ヒル関数をfittingさせることにより,n2,n3,K2,K3が求まる. We have obtained data for our newly devised hybrid promoter. By Hill function fitting, we have determined n2, n3, K2, and K3.
Expression6-1.JPG
Expression6-2.JPG

導出に関してはfor more detail.

Assays for hybrid promoter

These assays were done
-to determine the characteristics of our hybrid promoter.
The data from here were used for computational analyses and simulations.

Repression and activation check with LacI and AHL

Purpose


check if AHL activates lux-lac hybrid promoter
check if lacI represses lux-lac hybrid promoter

Result & Conclusion


AND gate by AHL & IPTG
Lux-lac hybrid promoter is activated only in the presence of AHL and IPTG.

IPTG assay

Purpose


To determine the order of the concentration of IPTG necessary for the activation of our lux-lac hybrid promoter in the LacI producing pTrc99A cells.
The order of the concentration is used for more detailed assay with narrower range of the IPTG concentration.

Result & Conclusion

2.Preliminary assays

We conducted preliminary assays to practice experiments and check if the materials and methods worked properly.
See more details