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Team:UCL London/From the lab/Expts
From 2009.igem.org
Contents |
Introduction
After the 4 stress detecting devices been constructed, our team conducted a series of experiments to test how they react to different stresses that can possibly occur during fermentation. The main focuses were on mis-folded protein, especially in periplasm, shear stress, and oxygen combustion levels of E.coli.
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K239000 Promoter degP] was thought to respond to mis-folded protein in the periplasm of E.coli. Various organic and inorganic chemicals which are suggested to cause protein denaturing in bacteria were used to induce protein mis-foldeding in the experiments. Important chemicals that were involved in our experiments are: Cu2+, Fe3+, NO3-, ethanol (C2H5OH), indole (C8H7N), 1-octanol (C8H18O) and toluene (C6H5CH3).
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K239001 Promoter spy] was thought to possibly detect high levels of shear stress during cultivation in a bioreactor. Additionally, spy showed response to some of the chemicals mentioned above as well.
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K239005 NarK Promoter] is switched on during anaerobic conditions (upregulated 100-fold and a further 8-fold in the presence of nitrate). Consequently, NarK was considered to detect low oxygen level, i.e. anaerobic metabolism. The difference between NarK promoter and mNarK promoter was tested by nitrate in the experiments illustrated below.
Copper 2+ ion & Ethanol Experiments
Aims
Procedures
