Tokyo/Works/Assay
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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. ]] | ||
'''Purpose''' | '''Purpose''' | ||
- | <br>check how LacI represses this hybrid promoter. In order to adjust LacI | + | <br>check how LacI represses this hybrid promoter. In order to adjust LacI repression<!--IPTGはLacIの濃度を調節しているわけではないのでconcentrationではなくrepressionに変更-->, IPTG was added. |
<br>These parameters(n3,K3) is decided. | <br>These parameters(n3,K3) is decided. | ||
<!--<br>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. | <!--<br>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. |
Revision as of 01:11, 26 October 2007
Works top 0.Hybrid promoter 1.Formulation 2.Assay1 3.Simulation 4.Assay2 5.Future works
Purpose of this assay 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 repression, IPTG was added.
These parameters(n3,K3) is decided.
Result & Conclusion
As the concentration of IPTG increases, GFP fluorescence increased. It indicates that the hybrid promoter in the cells expressing LacI became increasingly strengthened by decline of 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 as practice experiments and check of materials and methods.
⇒See more details
Before(Formulation) << Assay1 >> Assay1 contents >>>> Simulation