Tokyo/Works/Assay0
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===Results and Conclusion=== | ===Results and Conclusion=== | ||
| - | [[Image:hybridgraph2.JPG|thumb|350px| '''Fig.1: '''Result]] | + | [[Image:hybridgraph2.JPG|thumb|350px| '''Fig.1: '''Result. Activities of the newly devised lux lac hybrid promoter (hy) were determined in the presence and absence of LacI (lacI(+) and lacI(-), respectively) and those of AHL (AHL(+) and AHL(-)).]] |
[[Image:hybridgraph.JPG|thumb|300px| '''Fig.2: '''Result summary]] | [[Image:hybridgraph.JPG|thumb|300px| '''Fig.2: '''Result summary]] | ||
Revision as of 04:33, 25 October 2007
Works top 0. Hybrid promoter 1. Formulation 2. Assay1 3. Simulation 4. Assay2 5. Future works
Purpose 1:
To check if LacI hybrid promoter is activated by AHL and repressed by LacI.
Purpose 2:
To obtain parameters of LacI hybrid promoter for computational simulation.
Samples:
1-1. pTrc99A + [Lux lac hybrid promoter – GFP] (LacI+) (AHL+)
1-2. pTrc99A + [Lux lac hybrid promoter – GFP] (LacI+) (AHL-)
2-1. pBR322 TetR + [Lux lac hybrid promoter – GFP] (lacI-) (AHL+)
2-2. pBR322 TetR + [Lux lac hybrid promoter – GFP] (lacI-) (AHL-)
3-1. pTrc99A + [placQI – GFP] (placQI is constitutive promoter) (Pos. con.) (AHL+)
3-2. pTrc99A + [placQI – GFP] (placQI is constitutive promoter) (Pos. con.) (AHL-)
4-1. pTrc99A + [LacI promoter – GFP] (Neg. con.) (AHL+)
4-2. pTrc99A + [LacI promoter – GFP] (Neg. con.) (AHL-)
5-1. pBR322 TetR + [LacI promoter – GFP] (Neg. con. check) (AHL+)
5-2. pBR322 TetR + [LacI promoter – GFP] (Neg. con. check) (AHL-)
Repressor LacI expression:
pTrc99A expresses LacI
pBR322 TetR does NOT express LacI
Antibiotics resistance:
pTrc99A gives ampicillin-resistance
pBR322 TetR gives ampicillin-resistance
[LacI hybrid promoter – GFP] gives kanamicin-resistance
[placQI – GFP] gives kanamicin-resistance
Procedure:
Results and Conclusion
