From 2007.igem.org

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.

⇒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