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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 | | + | <span/ id="bottom">[https://2009.igem.org/ iGEM ] > [[Team:Paris#top | Paris]] > [[Team:Paris/Transduction_overview#top | Receiving the message]] > [[Team:Paris/Transduction_overview#bottom | Our strategy]] |
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| - | + | ==B. Our Strategy== | |
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| - | + | ===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 . | 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 . | ||
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'''problems :''' the message is unidirectionnal and unrepeatable. It would just be a proof of principle that a vesicle-mediated controlled communication is possible. | '''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. | We could ,with the help of Alfonso Jaramillo, design a synthetic PBP which could detect the substrate we choosed and activate a specific HK. | ||
Revision as of 22:09, 14 October 2009
iGEM > Paris > Receiving the message > Our strategy
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.
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.
