IGEM:MIT/2005/Receiver 1: ToxR

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Receiver Device #1 -- ToxR

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wiki-Will 8/18 - i'm out of town today and maybe a few more days. Possibly through the end of the week. the parents just sort of showed up!

Function

To cause gene expression in the presence of input signal using the ToxR pathway.

Device Depiction

Image:SystemToxRpathway.jpg Image:Receiver.1.ToxR e.ppt

Information path ...

  1. Fluoroscein introduced to system (as Fluoroscein+DNAspacer+Fluoroscein), travels into periplasm.
  2. Fluoroscein binds to adjacent transmembrane ToxR'-scFv fusions .
    1. Causes ToxR' dimerization
  3. GFP expressed @ CTX_promoter
    1. As a function of ToxR dimerization

People path ...

  1. Maxine gives us Fluoroscein, or identifies something else
  2. Maxine's antigen binds to Jenny's scFv
  3. Then, Jenny's scFv induces Will's ToxR' to dimerize
  4. The dimerization leads to the expression of Jessica's output @ CTXpromoter [cI, maybe]
  5. Ray makes the whole system look like "we meant to do that."
  6. Jen.

Device Parts

                Composite Parts for the first run of tests. 
                BBa_J07024	Device	ToxR'::scFv Fusion 1 (with promoter and rbs and terminator)	1648	
	  	BBa_J07025	Device	ToxR'::scFv Fusion 2 (with promoter and rbs and terminator)	1648	
	  	BBa_J07026	Device	ToxR'::scFv Fusion 3 (with promoter and rbs and terminator)	1648	
	  	BBa_J07027	Measurement	ToxR'::PhoA (with promoter and RBS and terminator)	2300	
	  	BBa_J07028	Measurement	ToxR'::MalE(with promoter and RBS and terminator)	2075

Experiments

Receiver 1 experiments: ToxR

  • Move contents to respective place.

Current Status

  1. In the process of verifying that we can get our signal into the periplasm (in lab)
  2. Part creation ...
    1. working out kinks in PhoA, malE isolation.
    2. soon ordering promotor-RBS tandem biobrick
  3. Continue research on putting scFv onto outermembrane of cell.

Open Issues/Questions

  1. The periplasm
    1. Is the signal going in?
    2. Are our scFv's going to come out (to the surface)?
  2. we won't know til we try! unless...

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