Saint Petersburg

From 2007.igem.org

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We intend to use the CusR promoter with the reporter gene to build sensor.
We intend to use the CusR promoter with the reporter gene to build sensor.
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'''Problem'''
 
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The main problem of our project is to make the "trigger" response with the two stable levels
 
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('''high copper сoncentration'''/'''high copper сoncentration''').
 
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Constructing such switch is a challenge.
 
[[Image:Saint-Petersburg_Trigger_invCu_sh.jpg|frame|left|Trigger with two copper-depended promoters]]
[[Image:Saint-Petersburg_Trigger_invCu_sh.jpg|frame|left|Trigger with two copper-depended promoters]]
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But this scheme is rather difficult and need many constructing operations.
But this scheme is rather difficult and need many constructing operations.
So, we decided to think more simple scheme, using the secondary metabolites.
So, we decided to think more simple scheme, using the secondary metabolites.
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'''Scheme with secondary metabolites'''
 
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[[Image:Saint-Petersburg_HSL-Activated_sh.jpg|frame|left|HSL-activating scheme]]
 
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Firsly, to use the secondary metabolites, we need to have metabolite-depended activator or repressor.
 
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Such biobrick avaliable in the iGem2007 registry as part <bbpart>BBa_C0062</bbpart>, the LuxR protein.
 
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In complex with HSL (homoserine lactone), LuxR binds to the Lux promoter, activating transcription  from part <bbpart>BBa_R0062</bbpart>.
 
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Secondary, we need a HSL-syntase. Probably, it may be part <bbpart>BBa_C0061</bbpart>, the enzyme for creating acyl-homoserine lactones from normal cell metabolites.
 
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If we have this enzyme after the copper-depended promoter, we can synthesize HSL in response to increasing copper level, and start transcription of LuxR promoter.
 
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In this scheme we can significantly increase signal level without making the triggers.
 
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For my mind, this sheme is optimal, certainly if this biobriks are working.
 

Revision as of 17:38, 17 July 2007

Saint-Petersburg iGem2007 team

This year our team is participating for the first time in iGem2007 project.

Our team works at the Saint-Petersburg State Polytechnical University in the department of Biophysics.

About Us

Our Project

General

The aim of our project is to build on the E.Coli bacteria the copper sensor. To solve this problem we need to use the copper-homeostasis system of E.Coli. The most convenient for our purposes protein is CusR, the component of the Copper-Responsive Two-Component System.

We intend to use the CusR promoter with the reporter gene to build sensor.

Trigger with two copper-depended promoters

So we perform a theoretical study of this problem.

So, if we want to use only proteins without secondary metabolites, the best trigger sheme is the next:

In this scheme we can observe the switching between two repressors (x and y), depending on the transcription level from the copper-depended promoter.

But this scheme is rather difficult and need many constructing operations. So, we decided to think more simple scheme, using the secondary metabolites.

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