Tokyo/Works/Assay0

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(Procedure:)
(Samples:)
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===Samples: ===
===Samples: ===
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<br>1. pTrc99A + [LacI hybrid promoter – GFP] (LacI+)
+
<br>1. pTrc99A + [Lux lac hybrid promoter – GFP] (LacI+)
-
<br>2. pBR322 TetR + [LacI hybrid promoter – GFP] (lacI-)
+
<br>2. pBR322 TetR + [Lux lac hybrid promoter – GFP] (lacI-)
-
<br>3. pTrc99A + [placQI – GFP] (placQI is constitutive promoter)
+
<br>3. pTrc99A + [placQI – GFP] (placQI is constitutive promoter) (Pos. con.)
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<br>4. pTrc99A + [LacI promoter – GFP]  
+
<br>4. pTrc99A + [LacI promoter – GFP] (Neg. con.)
-
<br>5. pBR322 TetR + [LacI promoter – GFP]  
+
<br>5. pBR322 TetR + [LacI promoter – GFP] (Neg. con. check)
<br>'''Repressor LacI expression:'''  
<br>'''Repressor LacI expression:'''  

Revision as of 04:20, 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. pTrc99A + [Lux lac hybrid promoter – GFP] (LacI+)
2. pBR322 TetR + [Lux lac hybrid promoter – GFP] (lacI-)
3. pTrc99A + [placQI – GFP] (placQI is constitutive promoter) (Pos. con.)
4. pTrc99A + [LacI promoter – GFP] (Neg. con.)
5. pBR322 TetR + [LacI promoter – GFP] (Neg. con. check)


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
Fig.2: Result summary