Tokyo/Works/Assay0

From 2007.igem.org

(Difference between revisions)
Line 2: Line 2:
<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: ===
-
===Procedure: ===
+
<br>1. pTrc99A + [LacI hybrid promoter – GFP] (LacI+)
-
===Result & Conclusion: ===
+
<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:


AHL assay Standard protocol

Fig.1: Result
Fig.2: Result summary