Tokyo/Works/Assay0
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<br>2-1. pBR322 TetR + [Lux lac hybrid promoter – GFP] '''(lacI-) (AHL+)''' | <br>2-1. pBR322 TetR + [Lux lac hybrid promoter – GFP] '''(lacI-) (AHL+)''' | ||
<br>2-2. pBR322 TetR + [Lux lac hybrid promoter – GFP] '''(lacI-) (AHL-)''' | <br>2-2. pBR322 TetR + [Lux lac hybrid promoter – GFP] '''(lacI-) (AHL-)''' | ||
- | <br>3. pTrc99A + [placQI – GFP] (placQI is constitutive promoter) | + | <br>3-1. pTrc99A + [placQI – GFP] (placQI is constitutive promoter) (Pos. con.) (AHL+) |
- | <br>4. pTrc99A + [LacI promoter – GFP] | + | <br>3-2. pTrc99A + [placQI – GFP] (placQI is constitutive promoter) (Pos. con.) (AHL-) |
- | <br>5. pBR322 TetR + [LacI promoter – GFP] (Neg. con. check) | + | <br>4-1. pTrc99A + [LacI promoter – GFP] (Neg. con.) (AHL+) |
+ | <br>4-2. pTrc99A + [LacI promoter – GFP] (Neg. con.) (AHL-) | ||
+ | <br>5-1. pBR322 TetR + [LacI promoter – GFP] (Neg. con. check) (AHL+) | ||
+ | <br>5-2. pBR322 TetR + [LacI promoter – GFP] (Neg. con. check) (AHL-) | ||
<br>'''Repressor LacI expression:''' | <br>'''Repressor LacI expression:''' |
Revision as of 04:28, 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