Difference between revisions of "Part:BBa K415031"

 
(7 intermediate revisions by 2 users not shown)
Line 4: Line 4:
 
This part is a promoter that was designed as a correction to the anomalous [[Part:BBa_R0065]] BioBrick. This part consists of a Lux Box binding site for the LuxR-C6HSL Complex; in addition, it contains two operator sites for the cI protein appropriated from the Lambda bacteriophage. Because basal expression without the LuxR-C6HSL complex present is expected to be minimal, and because the cI operator sites are positioned so as to block the critical -10 transcription initiation site on the promoter, if cI is high, the promoter is off; if AHL is low and cI is absent, the promoter is leaky; if AHL is high and cI is absent, the promoter is on.
 
This part is a promoter that was designed as a correction to the anomalous [[Part:BBa_R0065]] BioBrick. This part consists of a Lux Box binding site for the LuxR-C6HSL Complex; in addition, it contains two operator sites for the cI protein appropriated from the Lambda bacteriophage. Because basal expression without the LuxR-C6HSL complex present is expected to be minimal, and because the cI operator sites are positioned so as to block the critical -10 transcription initiation site on the promoter, if cI is high, the promoter is off; if AHL is low and cI is absent, the promoter is leaky; if AHL is high and cI is absent, the promoter is on.
  
Slightly stronger binding than [[Part:BBa_K415032]].
+
Slightly stronger binding than [[Part:BBa_K415032]]. For proof of reduced leakiness, see [[Part:BBa_K415032]].
 
+
{|style="text-align: center; border: 1px solid black;"
+
|-
+
|== K415031 is less leaky than R0065 ==
+
|-
+
|[[Image:Leakiness.png]]
+
|-
+
|The above images are FACS scatter plots of co-transformations of either pTSMa ([[Part:BBa_K415300]]) and [[Part:BBa_K415069]] or pTSMa and [[Part:BBa_K415023]]. The x-axis represents the amount of green fluorescence, and the y-axis represents the amount of red fluorescence. The parts K415069 and K415023 differ only in a single promoter sequence: K415069 has the K415031 promoter in place of a single R0065 promoter in the K415023 sequence. In this way, the more "leaky" the promoter, the more red fluorescing cells.
+
As you can see in the images, fewer cells with pTSMa+K415069 are leaky: 3% of total cells with pTSMa+K415069 fluoresce red, and 15% of total cells with pTSMa+K415023 fluoresce red.
+
|}
+
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 06:21, 27 October 2010

Improved PLux/cI-OR Promoter

This part is a promoter that was designed as a correction to the anomalous Part:BBa_R0065 BioBrick. This part consists of a Lux Box binding site for the LuxR-C6HSL Complex; in addition, it contains two operator sites for the cI protein appropriated from the Lambda bacteriophage. Because basal expression without the LuxR-C6HSL complex present is expected to be minimal, and because the cI operator sites are positioned so as to block the critical -10 transcription initiation site on the promoter, if cI is high, the promoter is off; if AHL is low and cI is absent, the promoter is leaky; if AHL is high and cI is absent, the promoter is on.

Slightly stronger binding than Part:BBa_K415032. For proof of reduced leakiness, see Part:BBa_K415032.

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
    COMPATIBLE WITH RFC[1000]