Difference between revisions of "Part:BBa K1541009"

Revision as of 03:33, 18 October 2014

sfGFP under promoter P(Rhl) with riboregulator RR12

Improved signal-to-noise ratio and decreased basal GFP expression (leakiness) due to the use of a riboregulator in combination with a quorum-sensing module. The fluorescence per OD₆₀₀ is shown for the LuxR-system with a complete riboregulator over an inducer-range of 10^-13 M to 10^-5 M (dashed, light blue). An incomplete riboregulator without the trans-activator shows the expected reduced sensitivity towards the inducer (dark blue). As a reference, a system with a non-regulated RBS (BBa_B0034) is shown (light blue). Data points are mean values of triplicate measurements in 96-well microtiter plates 200 min after induction ± standard deviation. For the full data set and kinetics please [http://2014.igem.org/Team:ETH_Zurich/contact contact] us or visit the [http://2014.igem.org/Team:ETH_Zurich/data/raw raw data] page.

this riboregulated promoter construct contains the quorum sensing promoter BBa_I14017 which can be activated in presence of RhlR (BBa_C0171) and C4-HSL, the product of the enzyme RhlI (BBa_C0170), succeeded by the gene for sfGFP. However, the promoter BBa_I14017 by itself shows some leakiness. Together with the ribogegulator 12^[18] the leakiness could be reduced.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal XhoI site found at 765
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 1: / Line 1: @@
 __NOTOC__
 <partinfo>BBa_K1541009 short</partinfo>
-[[File:ETH_Zurich_2014_LuxWith_and_WORibo.png|thumb|350px|right]]
+[[File:ETH_Zurich_2014_LuxWith_and_WORibo.png|thumb|350px|right|Improved signal-to-noise ratio and decreased basal GFP expression (leakiness) due to the use of a riboregulator in combination with a quorum-sensing module. The fluorescence per OD<sub>600</sub> is shown for the LuxR-system with a complete riboregulator over an inducer-range of 10<sup>-13</sup> M to 10<sup>-5</sup> M (dashed, light blue). An incomplete riboregulator without the ''trans''-activator shows the expected reduced sensitivity towards the inducer (dark blue). As a reference, a system with a [https://parts.igem.org/Part:BBa_B0034 non-regulated RBS (BBa_B0034)] is shown (light blue). Data points are mean values of triplicate measurements in 96-well microtiter plates 200 min after induction &plusmn; standard deviation. For the full data set and kinetics please [http://2014.igem.org/Team:ETH_Zurich/contact contact] us or visit the [http://2014.igem.org/Team:ETH_Zurich/data/raw raw data] page.]]
 [[File:ETH_Zurich_2014_LuxWith_and_WOSites.png|thumb|350px|right]]
 this riboregulated promoter construct contains the quorum sensing promoter [https://parts.igem.org/Part:BBa_I14017 BBa_I14017] which can be activated in presence of [https://parts.igem.org/Part:BBa_C0171:Experience RhlR (BBa_C0171)] and C4-HSL, the product of the enzyme [https://parts.igem.org/Part:BBa_C0170 RhlI (BBa_C0170)], succeeded by the gene for sfGFP. However, the promoter [https://parts.igem.org/Part:BBa_I14017 BBa_I14017] by itself shows some leakiness. Together with the ribogegulator 12<sup>[[Team:ETH_Zurich/project/references#refCallura|[18]]]</sup> the leakiness could be reduced.