Team:Paris/Transduction overview2 strategy

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<span/ id="bottom">[https://2009.igem.org/ iGEM ] > [[Team:Paris#top | Paris]] > [[Team:Paris/Transduction_overview#top | Reception]] > [[Team:Paris/Transduction_overview#bottom | Transduction]]
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== Overview  ==
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* [[Team:Paris/Transduction_overview2#Overview#bottom | Introduction]]
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* [[Team:Paris/Transduction_overview_transduction#bottom |A. Transduction]]
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** [[Team:Paris/Transduction_overview_transduction#B.1 ABC transporters|A.1 ABC transporters]]
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** [[Team:Paris/Transduction_overview_transduction#B.2 Two-component system|A.2  Two Component System]]
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* [[Team:Paris/Transduction_overview2_strategy#bottom |B. Our strategy]]
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** [[Team:Paris/Transduction_overview2_strategy#C.1 The Fec operon |B.1 The Fec operon]]
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** [[Team:Paris/Transduction_overview2_strategy#C.2 The trick TCS|B.2 The trick TCS]]
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* [[Team:Paris/Transduction_overview2_construction#bottom |C. Construction]]
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===B. Our Strategy===
===B. Our Strategy===
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Revision as of 14:43, 12 October 2009

iGEM > Paris > Reception > Transduction

Contents

Overview


B. Our Strategy

B.1 The Fec operon

Few ABC transporter such as FecABCD (iron transporter) are able to induce a response regardless of the tranlocation, due to the activity of FecA, moreover some mutant can also have a constitutive expression of FecABCD .


The plan would be to use FecA- mutant receiver and FecA+ mutant donor to transfert the constitutive FecA protein to the receiver. In this case the receiver will express the FecABCD operon without being induce by ferric citrate in the medium , and so we could place under the control of the Fec ABCD promoter, which is called pfec, the gene sequence encoding for the response. For the moment a response that would be easy to detect is the fluorescence of the RFP and the biobrick BBa-J61002 is the perfect candidate to test the system.


We also discovered that some fecR and fecI mutants can be use to amplify the signal because they have a constitutive activity. So we put under the control of pfec a FecR and FecI mutated. When they will be expressed, they will be activators of pfec and consequently of RFP. Normaly we would be able to obtain a increasing fluorescence.


fec operon induction


problems : the message is unidirectionnal and unrepeatable. It would just be a proof of principle that a vesicle-mediated controlled communication is possible.

B.2 The trick TCS

We could ,with the help of Alfonso Jaramillo, design a synthetic PBP which could detect the substrate we choosed and activate a specific HK. The previous work done by Valencia in 2006 was to design vanilin-sensitive PBP and a network for a graduated response whereas we just need a proteic sensitive PBP and a binary type of response.

Even if the idea of creating a synthethic protein was very attractive, it would have been difficult to have concrete result in just 3 months.

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