Team:Paris/Transduction overview strategy

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Overview

C. Our Strategy

C. 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.

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

C.x 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 (Valencia 2006) design a vanilin-sensitive PBP and a network for a graduated response whereas we just need a proteic sensitive PBP and a binary type of response.

strain and part needed: strain : E.coli MRi7 (unable to translocate ribose from periplasm to cytoplasme)

part :

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-====C.x Fec====
+====C. 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, some mutant can also have a constitutive expression of FecABCD . We could use FecA- mutant  receiver
+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 .
-and FecA+ mutant donor to transfert working FecA to the receiver. In this case the receiver will express the FecABCD operon, and so we could place under the control of the Fec ABCD promoter the gene sequence encoding for the response.
+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.