Tokyo/Expression level check

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(Difference between revisions)
(Result & Conclusion:)
(Result & Conclusion:)
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====Result & Conclusion:====
====Result & Conclusion:====
Two plasmid sets, A4Δp+pc1-GFP and A4 hybrid+GFP PBR322TetR (+)AHL, shows almost the same fluorescence of GFP, indicating that expression levels of both sets are almost the same though the latter is a bit smaller.
Two plasmid sets, A4Δp+pc1-GFP and A4 hybrid+GFP PBR322TetR (+)AHL, shows almost the same fluorescence of GFP, indicating that expression levels of both sets are almost the same though the latter is a bit smaller.
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<br>From the results the ranges of bistability and those of mono-stability are calculated as shown in Fig. 1.
+
<br>Based on the results, the ranges of bistability and those of mono-stability are calculated as shown in Fig. 1.
<br>.[[Image:pC1GFP.jpg|thumb|350px| '''Fig.1''' Genes downstream of the promoters on the both sides were expressed to almost the same degrees in terms of GFP fluorescence.|left]]
<br>.[[Image:pC1GFP.jpg|thumb|350px| '''Fig.1''' Genes downstream of the promoters on the both sides were expressed to almost the same degrees in terms of GFP fluorescence.|left]]
[[Image:Parameter set.jpg|thumb|500px|'''Fig.2 Calculated coexistance stable region'''|center|left]]
[[Image:Parameter set.jpg|thumb|500px|'''Fig.2 Calculated coexistance stable region'''|center|left]]

Revision as of 23:09, 25 October 2007


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

Activation check by cell-produced AHL   Expression level check on promoters + plasmid sets of A and B sides

Expression level check on two different promoters + plasmid sets

Purpose:

To measure and compare the activities of two different promoters + plasmid sets by fluorescence of GFP downstream of each promoter. Lambda cI-regulated promoter and the lux lac hybrid promoter were tested.

Samples:

・A4Δp pc1-GFP
・A4 hybrid GFP PBR322TetR (+)AHL
・A4 hybrid GFP PBR322TetR (-)AHL
*The promoter pcI sequence was designed to contain OR1, -35, and -10 regions, but not R2 or OR3.

Procedure:

AHL assay Standard protocol
Wash
OD and fluorescence were measured 0, 3, and 6 hours after the fresh culture incubation started.

Result & Conclusion:

Two plasmid sets, A4Δp+pc1-GFP and A4 hybrid+GFP PBR322TetR (+)AHL, shows almost the same fluorescence of GFP, indicating that expression levels of both sets are almost the same though the latter is a bit smaller.
Based on the results, the ranges of bistability and those of mono-stability are calculated as shown in Fig. 1.


.
Fig.1 Genes downstream of the promoters on the both sides were expressed to almost the same degrees in terms of GFP fluorescence.
Fig.2 Calculated coexistance stable region