IGEM:IMPERIAL/2007/Projects/Reporters: Difference between revisions

Revision as of 06:52, 27 July 2007

Description: Analyze the activity of a promoter (in terms of expression of a gene under that promoter) either in a quantitative way or through visualization of its activity in different tissues. The technique is based on beta-glucuronidase, an enzyme from the bacterium Escherichia coli. This enzyme when incubated with some specific colorless or not fluorescent substrates, can transform them into coloured or fluorescent products.
Potential use: Fluorescent output
Link: http://en.wikipedia.org/wiki/GUS_reporter_system

Enzyme: EC 1.13.12.7
Reaction: Photinus luciferin + O₂ + ATP <=> oxidized Photinus luciferin + CO₂ + AMP + diphosphate + light
Response Time: ~30 minutes
Steady State Time: 4 hours
Concentration sensitivity: Moderate
Advantages:
- Fast response
Disadvantages:

Gene name: luxCDABE
Description: 5 subunit protein which oxidizes FMN with the help of oxygen to generate FMN and H₂ complex. This complex breaks down to form light.
Enzyme: EC 1.14.14.3
Reaction: RCHO + reduced FMN + O₂ <=> RCOOH + FMN + H₂O + light
Response Time: ~30 minutes
Steady State Time: 8 hours
Concentration sensitivity: Moderate
Advantages:
- Fast response
Disadvantages:
- Five subunits required
- Metabolic stress, FMN is consumed
- Substrate limiting

Questions: What substrate does this need?

@@ Line 7: / Line 7: @@
 ===Firefly luciferase===
 *Gene name: ''lucff''
-*'''Description:'''
+*Description:
-*
+*Enzyme: [http://www.expasy.org/enzyme/1.13.12.7 EC 1.13.12.7]
+*Reaction: Photinus luciferin + O<sub>2</sub> + ATP <=> oxidized Photinus luciferin + CO<sub>2</sub> + AMP + diphosphate + light
+*Response Time: ~30 minutes
+*Steady State Time: 4 hours
+*Concentration sensitivity: Moderate
+*Advantages:
+**Fast response
+*Disadvantages:
+**
+**
 ===Bacterial luciferase===