Tokyo/Works/Assay
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<br>[[Tokyo/Works|Works top]] 0.[[Tokyo/Works/Hybrid promoter|Hybrid promoter]] 1.[[Tokyo/Works/Formulation |Formulation]] 2.[[Tokyo/Works/Assay |Assay1]] 3.[[Tokyo/Works/Simulation |Simulation]] 4.[[Tokyo/Works/Assay2 |Assay2]] 5.[[Tokyo/Works/Future works |Future works]] | <br>[[Tokyo/Works|Works top]] 0.[[Tokyo/Works/Hybrid promoter|Hybrid promoter]] 1.[[Tokyo/Works/Formulation |Formulation]] 2.[[Tokyo/Works/Assay |Assay1]] 3.[[Tokyo/Works/Simulation |Simulation]] 4.[[Tokyo/Works/Assay2 |Assay2]] 5.[[Tokyo/Works/Future works |Future works]] | ||
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| + | [[Tokyo/Hill function fitting |Purpose of this assay (Hill function fitting) ]] [[Tokyo/AHL assay|1.AHL assay]] [[Tokyo/IPTG assay|2.IPTG assay]] [[Tokyo/Preliminary assays |Preliminary assays]] | ||
| - | =[[Tokyo/Hill function fitting |Purpose of this assay (Hill function fitting) ]] = | + | ==[[Tokyo/Hill function fitting |Purpose of this 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． | <!--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.--> | ヒル関数をfittingさせることにより，n2,n3,K2,K3が求まる.--> | ||
Revision as of 07:02, 25 October 2007
Works top 0.Hybrid promoter 1.Formulation 2.Assay1 3.Simulation 4.Assay2 5.Future works
Purpose of this assay (Hill function fitting) 1.AHL assay 2.IPTG assay Preliminary assays
Purpose of this assay (Hill function fitting)
We have obtained data for our newly devised hybrid promoter. By Hill function fitting, we have determined n2, n3, K2, and K3.
⇒see more details
1.AHL assay
Purpose
check how AHL activates lux-lac hybrid promoter.
These parameters(n2,K2) is decided.
Result & Conclusion
As the concentration of AHL increases, GFP fluorescence increased, indicating that the hybrid promoter’s activation is strengthend with increasing concentration of AHL. From the activation graph in Fig. 1, the characteristics of the hybrid promoter expressed in Hill function, such as (n2,K2), is determined.
⇒see more details
2.IPTG assay
Purpose
check how LacI represses this hybrid promoter. In order to adjust LacI concentration, IPTG was added.
These parameters(n3,K3) is decided.
Result & Conclusion
As the concentration of IPTG increases, GFP fluorescence increased, indicating that the hybrid promoter in the LacI expressing cell became increasingly strengthened by decreasing repression by LacI. The activation graph in Fig. 2, the characteristics of the hybrid promoter expressed in Hill function is determined.
⇒see more details
Appendix:Preliminary assays
We conducted preliminary assays to practice experiments and check if the materials and methods worked properly.
⇒See more details
Before(Formulation) << Assay1 >> Assay1 contents >>>> Simulation
