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

Revision as of 06:49, 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

Gene name: luxCDABE
Description: 5 subunit protein
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 13: / Line 13: @@
 ===Bacterial luciferase===
 *Gene name: ''luxCDABE''
+*Description: 5 subunit protein
 *Enzyme: [http://www.expasy.org/enzyme/1.14.14.3 EC 1.14.14.3]
 *Reaction: RCHO + reduced FMN + O<sub>2</sub> <=> RCOOH + FMN + H<sub>2</sub>O + light
 *Response Time: ~30 minutes
-*Peak Response Time: ?
+*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?