Difference between revisions of "Part:BBa K091146:Experience"

Revision as of 18:38, 15 May 2009

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Applications of BBa_K091146

User Reviews

UNIQe526b0d60514794f-partinfo-00000000-QINU

No review score entered. Wideloache

This part was tested with the autoinducer molecules 3OC6 and 3OC12 in cells that constitutively expressed LuxR and LasR. It was expected that GFP would be expressed when 3OC12 was given to the cells but not when both 3OC12 and 3OC6 was given. This was because of the lux box that was put in the -35 to -10 region of the promoter. Binding of LuxR+3OC6 to this region was expected to result in repression of the promoter. I found that 3OC12 appeared to activate LuxR, which resulted in unexpected behavior. See the graphs below for my data.

This graph shows how the pLasLux promoter (K091146) is induced by 3OC12 and 3OC6. The pLas' promoter corresponds to BBa_K091117 and has no lux box in the -35 to -10 region of the promoter. The cell strain MC4100::K091206 expresses LuxR and LasR, while JM109 does not. I would have expected to see induction of pLasLux when only 3OC12 was added. We expected that the lack of induction is due to cross reactivity between 3OC12 and LuxR.

The Lux Receiver corresponds to K09100. The Las Receiver corresponds to K091134. This is evidence of cross reactivity between 3OC12 and LuxR, as 3OC12 is shown to induce the Lux Receiver.

Further discussion of these results can be found [http://www.bio.davidson.edu/courses/genomics/2008/DeLoache/HonorsThesis.pdf here]. The raw data can be downloaded [http://gcat.davidson.edu/GcatWiki/images/c/cb/FluorescenceExperiment.xls here].

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UNIQe526b0d60514794f-partinfo-00000002-QINU

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 This part was tested with the autoinducer molecules 3OC6 and 3OC12 in cells that constitutively expressed LuxR and LasR. It was expected that GFP would be expressed when 3OC12 was given to the cells but not when both 3OC12 and 3OC6 was given. This was because of the lux box that was put in the -35 to -10 region of the promoter. Binding of LuxR+3OC6 to this region was expected to result in repression of the promoter. I found that 3OC12 appeared to activate LuxR, which resulted in unexpected behavior. See the graphs below for my data.
-[[Image:Graph1 WD.png|400px|thumb|none|This gel shows how the pLasLux promoter (K091146) is induced by 3OC12 and 3OC6. The pLas' promoter corresponds to BBa_K091117 and has no lux box in the -35 to -10 region of the promoter. The cell strain MC4100::K091206 expresses LuxR and LasR, while JM109 does not. I would have expected to see induction of pLasLux when only 3OC12 was added. We expected that the lack of induction is due to cross reactivity between 3OC12 and LuxR.]]
+[[Image:Graph1 WD.png|400px|thumb|none|This graph shows how the pLasLux promoter (K091146) is induced by 3OC12 and 3OC6. The pLas' promoter corresponds to BBa_K091117 and has no lux box in the -35 to -10 region of the promoter. The cell strain MC4100::K091206 expresses LuxR and LasR, while JM109 does not. I would have expected to see induction of pLasLux when only 3OC12 was added. We expected that the lack of induction is due to cross reactivity between 3OC12 and LuxR.]]
 [[Image:Graph2 WD.png|400px|thumb|none|The Lux Receiver corresponds to K09100. The Las Receiver corresponds to K091134. This is evidence of cross reactivity between 3OC12 and LuxR, as 3OC12 is shown to induce the Lux Receiver.]]
 Further discussion of these results can be found [http://www.bio.davidson.edu/courses/genomics/2008/DeLoache/HonorsThesis.pdf here]. The raw data can be downloaded [http://gcat.davidson.edu/GcatWiki/images/c/cb/FluorescenceExperiment.xls here].