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 | | + | [[Tokyo/Hill function fitting |Objective of this assay]] [[Tokyo/AHL assay|Effect of AHL]] [[Tokyo/IPTG assay|Effect of IPTG]] [[Tokyo/Preliminary assays |Preliminary assays]] |
| - | ==[[Tokyo/Hill function fitting | | + | ==[[Tokyo/Hill function fitting |Objective 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.--> | ||
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== [[Tokyo/AHL assay|Effect of AHL]] == | == [[Tokyo/AHL assay|Effect of AHL]] == | ||
<br>[[Image:hill AHL.JPG|thumb|210px|'''Fig.1 The result of AHL assay''' Fluorescence intensity (arbitrary unit, a.u.) as a function of the concentration of AHL was determined. ]] | <br>[[Image:hill AHL.JPG|thumb|210px|'''Fig.1 The result of AHL assay''' Fluorescence intensity (arbitrary unit, a.u.) as a function of the concentration of AHL was determined. ]] | ||
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Bytesting how AHL activates lux-lac hybrid promoter, parameters(n2,K2) should be determined. | Bytesting how AHL activates lux-lac hybrid promoter, parameters(n2,K2) should be determined. | ||
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[[Image:hill IPTG.JPG|thumb|210px|'''Fig.2 The result of IPTG assay''' Fluorescence intensity as a function of the concentration of IPTG was determined. ]] | [[Image:hill IPTG.JPG|thumb|210px|'''Fig.2 The result of IPTG assay''' Fluorescence intensity as a function of the concentration of IPTG was determined. ]] | ||
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By testing how LacI represses lux-lac hybrid promoter, parameters(n3,K3) should be decided. In order to adjust effective concentration of LacI, IPTG was added. | By testing how LacI represses lux-lac hybrid promoter, parameters(n3,K3) should be decided. In order to adjust effective concentration of LacI, IPTG was added. | ||
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Revision as of 18:46, 26 October 2007
Works top 0.Hybrid promoter 1.Formulation 2.Assay1 3.Simulation 4.Assay2 5.Future works
Objective of this assay Effect of AHL Effect of IPTG Preliminary assays
Objective 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
Effect of AHL
Objective
Bytesting how AHL activates lux-lac hybrid promoter, parameters(n2,K2) should be determined.
Result & Conclusion
GFP fluorescence increased in AHL-dose dependent manner, indicating that the hybrid promoter is enhanced by LuxR via AHL. Taken together with the graph shown in fig.1and formulation of Hill function fitting described above , the characteristics of the hybrid promoter expressed in Hill function, such as (n2,K2), is determined.
⇒see more details
2.IPTG assay
Objective
By testing how LacI represses lux-lac hybrid promoter, parameters(n3,K3) should be decided. In order to adjust effective concentration of LacI, IPTG was added.
Result & Conclusion
GFP fluorescence intensity was enhanced in IPTG-dose dependent manner. It indicates that the LacI repression was getting weak by adding IPTG. Taken together with the graph shown in Fig. 2 and the formulation of Hill function fitting described above, the characteristics of the hybrid promoter expressed in Hill function was determined.
⇒see more details
Appendix: Preliminary assays
We conducted preliminary assays as practice experiments and check of materials and methods.
⇒See more details
Before(Formulation) << Assay1 >> Assay1 contents >>>> Simulation
