Difference between revisions of "Part:BBa K415032"

Latest revision as of 06:20, 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 weaker binding than Part:BBa_K415031.

K415032 is less leaky than R0065

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.

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]

@@ 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.
-Slightly weaker binding than [[Part:BBa_K415031]]. For proof of reduced leakiness, see [[Part:BBa_K415031]].
+Slightly weaker binding than [[Part:BBa_K415031]].
+{|style="text-align: center; border: 1px solid black;"
+|-
+!
+== K415032 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