"

From 2013.igem.org

NarX

 

NarX

Nitrate sensing: proof of concept
NarX is the histidine kinase
NarL is the response regulator
http://www.sciencedirect.com/science/article/pii/S0003269704000910
http://partsregistry.org/Part:BBa_M33030
http://onlinelibrary.wiley.com/doi/10.1046/j.1472-765X.1997.00064.x/abstract
Nitrate and Nitrite hybrid sensing protein (NarX + Tar)http://jb.asm.org/content/188/11/3944.full.pdf+html
this is what an old igem team did:
https://2009.igem.org/Team:Edinburgh. They used NsrR
Didnt have a chimera - that is the difference
Do like 14-152 or 14-174
http://www.uniprot.org/uniprot/P0AFA2

Amino Acid Sequence:
QVALIVLLSTAIGLAGMAVSGWLVQGVQGSAHAINKAGSLRMQSYRLLAAVPLSEKDKPLIKEMEQTAFSAELTRAAERDGQLAQLQ

GLQDYWRNELIPALMRAQNRETVSADVSQFVAGLDQLVSGFDRTTEMRIETVVLVHRVMAVFMALLLVFTIIWL
or

QVALIVLLSTAIGLAGMAVSGWLVQGVQGSAHAINKAGSLRMQSYRLLAAVPLSEKDKPLIKEMEQTAFSAELTRAAERDGQLAQLQ

GLQDYWRNELIPALMRAQNRETVSADVSQFVAGLDQLVSGFD

Experiment Procedure:

Add nitrate to the cell suspension.
The positive control will be...
The negative control will be no nitrate in the suspension.

Used a narK E. coli mutant which does not transport nitrate across the membrane.  Adding octylglucoside to the cell suspension increased

membrane permeability for NO3.  When it was transported into the cell it was reduced to NO2.  Adding N3 caused decreased NO3 and

thus nitrate reductase activity.

Nitrate uptake was sensed with nitrate ion selective electrodes.
http://www.sciencedirect.com/science/article/pii/0014579389809068

https://en.wikipedia.org/wiki/Ion_selective_electrode

narK transport protein gene.  Used only during anaerobic growth in the presence of nitrate.
http://ecocyc.org/ECOLI/NEW-IMAGE?type=GENE&object=EG10642

Octylglucoside
http://en.wikipedia.org/wiki/Octyl_glucoside

This is where the deatils of NarX will go. This is where the deatils of NarX will go. This is where the deatils of NarX will go. This is where the deatils of NarX will go. This is where the deatils of NarX will go. This is where the deatils of NarX will go. This is where the deatils of NarX will go. This is where the deatils of NarX will go. This is where the deatils of NarX will go.This is where the deatils of NarX will go. This is where the deatils of NarX will go. This is where the deatils of NarX will go. This is where the deatils of NarX will go. This is where the deatils of NarX will go. This is where the deatils of NarX will go.

Gel samples

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Flow sample

 

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