Team:Grenoble/Biology/Notebook/June/week 26


iGEM Grenoble 2012



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Week 26: June 25th to July 01st

For the detection module, we decided to work on a modified membrane receptor. It consists of two merged E. coli membrane receptors. The extracellular part is the extracellular part of Tap, a dipeptide membrane receptor. The cytoplasmic part is the cytoplasmic part of the EnvZ receptor, which has the ability to activate OmpR (by phosphorilation) which is a transcription factor.


It will be a proof of concept for a bigger system which is a complete pathogene detection module :

1st network : with amplifier 1 (RsmA-rsmY)

Amplifier 1 : RsmA-rsmY system characterisation: (3 final plasmids)

Plasmid Mapping

On a pSB4K5 plasmid, we wanted to put the construction: pLAC_fha1_eCFP
On a pSB3C5 plasmid, we wanted to put the construction: pLAC_RBS_RsmA
On a pSB1A3 plasmid, we wanted to put the construction: pLAC_rsmY

Biobricks involved

pLAC_RBS (BBa_I13601) => final length ≃ 90bp
pLAC (BBa_I13601) => final length ≃ 90bp
eCFP (BBa_E0422 or BBa_E0022) => final length ≃ 800bp
plasmid pSB4K5 => final length ≃ 2400bp
plasmid pSB3C5 => final length ≃ 2400bp
plasmid pSB1A3 => final length ≃ 2400bp

New parts

RsmA (iGEM Grenoble 2011) => final length ≃ 200bp
rsmY (iGEM Grenoble 2011) => final length ≃ 170bp
fha1 (iGEM Grenoble 2011) => final length ≃ 80bp

2nd network : with amplifier 2 (cAMP-CRP)

Amplifier 2: cAMP-CRP system (first construction): (1 final plasmid)

Plasmid Mapping

On the pSB4K5 plasmid, we wanted to put the construction: pAra/Bad_RBS_GFP_RBS_Cya

Biobricks involved

pSB4K5 plasmid => final length ≃ 2400bp

New parts

pAra/Bad_RBS_GFP (Alon collection) => final length ≃ 1300bp
RBS_Cya (E. coli chromosom) => final length ≃ 2600bp