Difference between revisions of "Part:BBa R0079:Experience"

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<partinfo>BBa_R0079 AddReview 4</partinfo>
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<I>ETH Zurich 2014</I>
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= Characterization of two-order crosstalk on the promoter =
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== Background information ==
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=== System considered ===
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=== Modeling crosstalk ===
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== First-order crosstalk ==
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=== First Level crosstalk: LasR binds to different HSL and activates the promoter ===
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[[File:ETH Zurich 1crosstalkPlas.png|400px|center]]
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=== Second Level crosstalk: other regulatory proteins, like LuxR, bind to their natural HSL substrate and activates the promoter ===
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[[File:ETH_Zurich_2crosstalkPlas.png|400px|center]]
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== Second order crosstalk: Combination of both cross-talk levels  ==
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Other regulatory proteins, like LuxR, bind to different HSL and activates the promoter
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[[File:ETh Zurich 3crosstalkPlas.png|400px|center]]
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== Results ==
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Revision as of 08:53, 20 October 2014

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_R0079

User Reviews

UNIQcc4d9875c18c807d-partinfo-00000000-QINU UNIQcc4d9875c18c807d-partinfo-00000001-QINU

••••

ETH Zurich 2014

Characterization of two-order crosstalk on the promoter

Background information

System considered

Modeling crosstalk

First-order crosstalk

First Level crosstalk: LasR binds to different HSL and activates the promoter

ETH Zurich 1crosstalkPlas.png

Second Level crosstalk: other regulatory proteins, like LuxR, bind to their natural HSL substrate and activates the promoter

ETH Zurich 2crosstalkPlas.png

Second order crosstalk: Combination of both cross-talk levels

Other regulatory proteins, like LuxR, bind to different HSL and activates the promoter

ETh Zurich 3crosstalkPlas.png

Results

No review score entered. NYMU-Taipei 2009

NYMU 2009-09-29.png We have characterised the strength of the promoters pCI, p22, pLux, pLas relative to pCI. More details are available at the [http://2009.igem.org/Team:NYMU-Taipei/Project/Promoter_Strength_Testing NYMU-Taipei iGEM09 wiki].
;

No review score entered. Northwestern 2011

The Northwestern iGEM team used this part as a unit within our Pseudomonas Aeruginosa biosensor. When this LasR/PAI regulated promoter is induced at varying concentrations of PAI in the presence of excess LasR, we observed GFP fluorescence in accordance with the graph below.


LasR graph.jpg

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•••

Tokyo-tech iGEM 2011

Judging from Northwestern iGEM 2011 team's data, in the presence of 3OC12-HSL, fluorescence intensity was about 3-fold higher than that in the absence of 3OC12-HSL.


We improved this part. lasI promoter(BBa_K649000) which we constructed was more successfully regulated by 3OC12-HSL. GFP expression after induction of 3OC12-HSL is 170 times as high as before.

Effect of 3OC12-HSL induction on fluorescence intensity
This work is done by Takuya Tsubaki.


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No review score entered. Tsinghua-A 2011

Tsinghua-A 2011 assembled E0840 (BBa_E0840) under the pLas promoter (BBa_R0079) that was contained into K574009 (BBa_K574009). We kept pSB1A2 as the scaffold vector.

K574009 chart.png
From the chart, we can see that cells were hardly induced in the control group, and with the concentration of inducer growing, the intensity of GFP increased by groups. The most efficient concentration of inducer was around 10^-5M, and higher concentration may lead to the expression of GFP decreasing. Additionally, to most groups, the intensity of GFP reached its maxium after 4 hours.
More details are available at the [http://2011.igem.org/Team:Tsinghua-A/Parts Tsinghua-A 2011 Wiki].


•••••

iGEM Dundee 2014

Dundee iGEM 2014 used this lasB promoter region to build a composite part BBa_K1315009. This was designed as a biosensor for Pseudomonas aeruginosa AutoInducer-1 (PAI-1), which was to be used in a bio-electronic device to improve diagnostics for Cystic Fibrosis patients. Details of experimental work are logged on the experience page of BBa_K1315009.