Difference between revisions of "Part:BBa K218011"
Line 22: Line 22:
 
The functionality of the reporter was tested by measuring the fluorscence of reporter together with LuxO D47E (K218017) mutant, and this fluorscence was compared to the fluorscence of our positive control (R0040 + I13500).  
 
The functionality of the reporter was tested by measuring the fluorscence of reporter together with LuxO D47E (K218017) mutant, and this fluorscence was compared to the fluorscence of our positive control (R0040 + I13500).  
 
The following is the protocol of the fluorescence reading.
 
The following is the protocol of the fluorescence reading.
 +
<br>
 
1. Power on the Bio-tec Synergy HT plate reader, or another plate reader, and KC4 application.
 
1. Power on the Bio-tec Synergy HT plate reader, or another plate reader, and KC4 application.
−
2. On a black 96 well plate, aliquot samples in required wells.
+
<br>2. On a black 96 well plate, aliquot samples in required wells.
−
3. Measure OD600 by using the following settings:
+
<br>3. Measure OD600 by using the following settings:
−
Reader: absorbance
+
<br>Reader: absorbance
−
Still editing.
+
<br>Still editing.

Revision as of 01:35, 21 October 2009

LuxO inducible GFP

LuxO when phosphorylated binds to Pqrr4, GFP is expressed. Alternatively, LuxO D47E mutant can also bind to the Pqrr4 and induce GFP production.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 945


Pictures

Characterization of the reporter (Pqrr4 + I13500) circuit

The functionality of the reporter was tested by measuring the fluorscence of reporter together with LuxO D47E (K218017) mutant, and this fluorscence was compared to the fluorscence of our positive control (R0040 + I13500). The following is the protocol of the fluorescence reading.
1. Power on the Bio-tec Synergy HT plate reader, or another plate reader, and KC4 application.
2. On a black 96 well plate, aliquot samples in required wells.
3. Measure OD600 by using the following settings:
Reader: absorbance
Still editing.