Team:Paris/Transduction overview2 construction

iGEM > Paris >  Receiving the message >  Construction

Construction
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a.menu_sub_active { padding-left: 7px; padding-right: 7px; color:#b0310e; font-weight:bold; }  Main |  Transduction|  Our strategy|  Construction

Proof of the activity of pfec
After all the process to obtain pfec as a biobrick, we have to clone it into BBa j61002 which allows to test promotor efficiency. After the transformation of this plasmid into a FecA + strains and on a medium were the ferric citrate is the only iron source, the RFP should be expressed and visible.

Proof of the activity of FecA
Once again after the process of obtaining FecA as a biobrick, we have to clone it into a low copy number plasmid such as psb3t5 and with an inducible promotor such as pBAD Afterwards it will be necessary to double transform it with the previous pfec construction in a FecA deleted strain and on a 1% glucose medium and 1% arabinose medium. So on the 1% arabinose medium the FecA would be expressed, translocated in the outer membrane and its constitutive activity will activated fecR and FecI and finaly pfec, and like previously the RFP will be expresed and visible. The glucose medium is a a negative control.

 

reference

 * 1) 1995- Cosima Harle & Volkmar Braun - Signal transfer through three compartments transcription initiation of the Escherichia coiferric citrate transport system from the cell surface
 * 2) 2000 - Stock & Goudreau – Two-component signal transduction
 * 3) 2001 - Mishima & Murata - Super-channel in bacteria function and structure of the macromolecule import system mediated by a pit-dependent ABC transporter
 * 4) 2009 - Akyriakidis & Tiligada – signal transduction TCS the AtoSC paradigm
 * 5) 2009 - Tomii & Kanehisa – comparative analysis of ABC transporter