Tokyo/Works/Assay0

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(Results and Conclusion)
(Results and Conclusion)
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===Results and Conclusion===
===Results and Conclusion===
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[[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(-)).]]
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[[Image:hybridgraph2.JPG|thumb|350px| '''Fig.1: '''Result of assay 0. 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 04:35, 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:


AHL assay Standard protocol

Results and Conclusion

Fig.1: Result of assay 0. 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(-)).
Fig.2: Result summary