Tokyo/Works/Assay0
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<br>[[Tokyo/Works|Works top]] 0. [[Tokyo/Works/Hybrid promoter|Hybrid promoter]] 1. [[Tokyo/Works/Formulation |Formulation]] 2. [[Tokyo/Works/Assay |Assay1]] 3. [[Tokyo/Works/Simulation |Simulation]] 4. [[Tokyo/Works/Assay2 |Assay2]] 5. [[Tokyo/Works/Future works |Future works]] | <br>[[Tokyo/Works|Works top]] 0. [[Tokyo/Works/Hybrid promoter|Hybrid promoter]] 1. [[Tokyo/Works/Formulation |Formulation]] 2. [[Tokyo/Works/Assay |Assay1]] 3. [[Tokyo/Works/Simulation |Simulation]] 4. [[Tokyo/Works/Assay2 |Assay2]] 5. [[Tokyo/Works/Future works |Future works]] | ||
| - | ===Purpose: === | + | ===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: === | ===Samples: === | ||
| - | + | <br>1. pTrc99A + [LacI hybrid promoter – GFP] (LacI+) | |
| - | === | + | <br>2. pBR322 TetR + [LacI hybrid promoter – GFP] (lacI-) |
| + | <br>3. pTrc99A + [placQI – GFP] (placQI is constitutive promoter) | ||
| + | <br>4. pTrc99A + [LacI promoter – GFP] | ||
| + | <br>5. pBR322 TetR + [LacI promoter – GFP] | ||
| + | |||
| + | <br>'''Repressor LacI expression:''' | ||
| + | <br>pTrc99A expresses LacI | ||
| + | <br>pBR322 TetR does NOT express LacI | ||
| + | |||
| + | <br>'''Antibiotics resistance:''' | ||
| + | <br>pTrc99A gives ampicillin-resistance | ||
| + | <br>pBR322 TetR gives ampicillin-resistance | ||
| + | <br>[LacI hybrid promoter – GFP] gives kanamicin-resistance | ||
| + | <br>[placQI – GFP] gives kanamicin-resistance | ||
| + | |||
| + | ===Procedure:=== | ||
| + | <br>[[Tokyo/AHL assay Standard protocol |AHL assay Standard protocol ]] | ||
[[Image:hybridgraph2.JPG|thumb|350px| '''Fig.1: '''Result]] | [[Image:hybridgraph2.JPG|thumb|350px| '''Fig.1: '''Result]] | ||
[[Image:hybridgraph.JPG|thumb|300px| '''Fig.2: '''Result summary]] | [[Image:hybridgraph.JPG|thumb|300px| '''Fig.2: '''Result summary]] | ||
Revision as of 10:37, 24 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 + [LacI hybrid promoter – GFP] (LacI+)
2. pBR322 TetR + [LacI hybrid promoter – GFP] (lacI-)
3. pTrc99A + [placQI – GFP] (placQI is constitutive promoter)
4. pTrc99A + [LacI promoter – GFP]
5. pBR322 TetR + [LacI promoter – GFP]
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:
