From 2009.igem.org
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| - | ===C. Our Strategy=== | + | ===B. Our Strategy=== |
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| - | ====C.1 The Fec operon ==== | + | ====B.1 ==== |
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| - | 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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| - | 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.
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| - | 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.
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| - | [[Image:Transduction_overview_fec.png|500px|center| fec operon induction]]
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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.
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| - | ====C.2 The trick TCS==== | + | |
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| - | We could ,with the help of Alfonso Jaramillo, design a synthetic PBP which could detect the substrate we choosed and activate a specific HK.
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| - | 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.
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| - | 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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Revision as of 14:52, 12 October 2009
iGEM > Paris > Reception > Our strategy
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