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From 2010.igem.org

Organisation and logistic

- flights in the US

- Discussion of possible Substances for detection

-> list of Substances:

2-Chlorphenol (drug), Capcaicin (spiciness), Dopamin and its derivates (human hormones),

2,4,6 Trichloranisol(Responsible for bad taste in red wine))

- Literature research for the virA sensor system

contact of reaserch groups in order to get an already working system (failed)

- Evaluation of the mutageneses strategy

-> Error Proune PCR, DNA shuffling, directed evolution, Protein coupling assay

- contact to local newspapers

Wetlab

accomplished:

- qRT-PCR of induced aggrobacterium tumefaciens c58

-> the strain could be significantly induced by acetosyringon

- Synthesis of the virG by MrGene (use without rpoA, clear of illegal restriction sites, codon usage for a.tumrfaciens)

- Testing of the Promega readout machine (GloMax multiplate reader) for the LUC-assay (working)

-> calibration of the GloMax

- Testing of the virA construct from another researchgroup

-> the virA construct could not be amplified

- Testing of the virA biobrick taken of the iGEM regestry

-> Correction and improvement of the virA biobrick

- cloning of a constitutive promotor and a rbs in front of the improved virA

to be done:

- cloning of virG into the right biobrick vector

- characterisation of new build standalone virG biobrick

- characterisation of new build virA biobrick

- cloning of the antibiotic resistence in the correct biobrick backbone

- cloning of the construct backbone (promotor, virA, terminator, virB, virG, readout)

- Error Proune PCR of virA

- Sensitivity test by antibiotic gradient

- Modelling