Team:Linkoping Sweden/Demonstrate

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All work done on BBa_K2671420

Plasmids used during the characterization of our biobrick

Figure x. Plasmids used in our experiments. pSub. can be either of mNG-Aß1-42, EGFP-Aß1-42, a-synuclein-EGFP and Tau0N4R-EGFP. As seen in the next figure, four combinations were introduced into E.coli (BL21). Substrate, substrate and GroES, substrate and GroES and GroE, substrate and GroE.

Overview of our experimental design to archive the results shown below

Figure x. A simple explaination on how we archived our experimental results. The plasmids shown is the same as before, where A can be either of mNG-Aß1-42, EGFP-Aß1-42, a-synuclein-EGFP and Tau0N4R-EGFP, B is always pGroE7 and C is pSB4A5-GroES. The concentrations used for inducing the chaperone plasmids was 0.25 mg/ml L-arabinose, 200 ng/ml tetracycline. For the substrate plasmids we used 0.5 mM IPTG, and the substrate was induced 30 mins after the chaperone plasmids.

Results for the mNG-Aß1-42 substrate protein

Figure x. Results for mNG-Aß1-42. Top graphs show fluorescene intensity divided by the start OD600 over 16 hours at 37 degrees. The bottom graphs show the normalized values from the top graphs, this creates a much better understanding and visual presentation of the kinetics of the substrate proteins folding rate in concert with the different chaperone combinations.

Results for the EGFP-Aß1-42 substrate protein

Results for the a-synuclein-EGFP substrate protein

Results for the Tau0N4R-EGFP substrate protein