Tokyo/Works/Assay

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


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.
Expression6-1.JPG
Expression6-10-2.JPGExpression6-11.JPG

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1.AHL assay


Fig.1

Purpose
check how AHL activates lux-lac hybrid promoter.
These parameters(n2,K2) is decided.

Result & Conclusion
Lux-lac hybrid promoter is activated by AHL as shown in Fig. 1.


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2.IPTG assay

Fig.2

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.

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Appendix:Preliminary assays

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

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Before(Formulation) << Assay1 >> Assay1 contents >>>> Simulation