Tokyo/Works/Assay

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__NOTOC__
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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]]
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<br>[[Tokyo/Works|Works top]]&nbsp;&nbsp;&nbsp;0.[[Tokyo/Works/Hybrid promoter|Hybrid promoter]]&nbsp;&nbsp;&nbsp;1.[[Tokyo/Works/Formulation |Formulation]]&nbsp;&nbsp;&nbsp;2.[[Tokyo/Works/Assay |Assay1]]&nbsp;&nbsp;&nbsp;3.[[Tokyo/Works/Simulation |Simulation]]&nbsp;&nbsp;&nbsp;4.[[Tokyo/Works/Assay2 |Assay2]]&nbsp;&nbsp;&nbsp;5.[[Tokyo/Works/Future works |Future works]]
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[[Tokyo/Hill function fitting |Objective of this assay]]　　[[Tokyo/AHL assay|Effect of AHL]]　　[[Tokyo/IPTG assay|Effect of LacI-IPTG]]　　[[Tokyo/Preliminary assays |Preliminary assays]]
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[[Tokyo/Hill function fitting |Objective of this assay]]&nbsp;&nbsp;&nbsp;[[Tokyo/AHL assay|Effect of AHL]]&nbsp;&nbsp;&nbsp;[[Tokyo/IPTG assay|Effect of LacI-IPTG]]　　[[Tokyo/Preliminary assays |Preliminary assays]]
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====⇒see more [[Tokyo/Hill function fitting|details]]====
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==== ==> see more [[Tokyo/Hill function fitting|details]]====
  <!--== Assays for hybrid promoter ==
  <!--== Assays for hybrid promoter ==
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<br>[[Image:hill AHL.JPG|thumb|230px|'''Fig. 1 Effect of AHL on hybrid promoter''' Enhanced fluorescence intensity represents up-regulation of LuxR via AHL.|left]]
<br>[[Image:hill AHL.JPG|thumb|230px|'''Fig. 1 Effect of AHL on hybrid promoter''' Enhanced fluorescence intensity represents up-regulation of LuxR via AHL.|left]]
'''Objective'''
'''Objective'''
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Bytesting how AHL activates lux-lac hybrid promoter,  parameters(n2,K2) should be determined.
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By testing how AHL activates lux-lac hybrid promoter,  parameters(n2,K2) should be determined.
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'''Result & Conclusion'''
'''Result & Conclusion'''
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<br>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.  
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<br>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.1 and formulation of Hill function fitting described above , the characteristics of the hybrid promoter expressed in Hill function, such as (n2,K2), are determined.  
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====⇒see more [[Tokyo/AHL assay|details]]====
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==== ==> see more [[Tokyo/AHL assay|details]]====
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[[Image:hill IPTG.JPG|thumb|215px|'''Fig. 2 Effect of LacI on hybrid promoter''' Enhanced fluorescence intensity represents down-regulation of LacI repression via IPTG.|left]]
[[Image:hill IPTG.JPG|thumb|215px|'''Fig. 2 Effect of LacI on hybrid promoter''' Enhanced fluorescence intensity represents down-regulation of LacI repression via IPTG.|left]]
'''Objective'''
'''Objective'''
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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.  
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By testing how LacI represses the 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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'''Result & Conclusion'''
'''Result & Conclusion'''
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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.
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GFP fluorescence intensity was enhanced in an 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.
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====⇒see more [[Tokyo/IPTG assay|details]]====
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==== ==> see more [[Tokyo/IPTG assay|details]]====
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We conducted preliminary assays as practice experiments and check of materials and methods.
We conducted preliminary assays as practice experiments and check of materials and methods.
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====⇒See more [[Tokyo/Preliminary assays |details]]====
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==== ==> See more [[Tokyo/Preliminary assays |details]]====
== ==
== ==
[[Tokyo/Works/Formulation |Before(Formulation)]]　<<　[[Tokyo/Works/Assay |Assay1]]　>>　[[Tokyo/Hill function fitting|Assay1 contents]]　>>>>　[[Tokyo/Works/Simulation |Simulation]]　　
[[Tokyo/Works/Formulation |Before(Formulation)]]　<<　[[Tokyo/Works/Assay |Assay1]]　>>　[[Tokyo/Hill function fitting|Assay1 contents]]　>>>>　[[Tokyo/Works/Simulation |Simulation]]　　

Latest revision as of 04:53, 27 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 LacI-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.


Siki1.JPG

Expression6-10-2.JPG
Siki2.JPG



==> see more details


Effect of AHL


Fig. 1 Effect of AHL on hybrid promoter Enhanced fluorescence intensity represents up-regulation of LuxR via AHL.

Objective By testing 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.1 and formulation of Hill function fitting described above , the characteristics of the hybrid promoter expressed in Hill function, such as (n2,K2), are determined.


==> see more details




Effect of LacI: IPTG assay

Fig. 2 Effect of LacI on hybrid promoter Enhanced fluorescence intensity represents down-regulation of LacI repression via IPTG.

Objective By testing how LacI represses the 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 an 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　　

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