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From 2010.igem.org

Contents 1 Organisation and logistic 2 Wetlab 2.1 accomplished 2.2 to be done

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; TV; Radio

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

• cloning of virG into the right biobrick vector

• creating new antibiotic biobricks

• Testing Top10 and Ec100D for ccdb-gene

to be done

• characterisation of new build standalone virG biobrick

• characterisation of new build virA biobrick

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

• Error Prone PCR of virA

• Sensitivity test by antibiotic gradient

• Modeling

• Constructing a new Backboneplasmid -> creating a cloning system in order to insert the r6k-origin in all psb-backbone plasmids

• Testing of the ccdb-death gene