Team:Linkoping Sweden/Demonstrate
Demonstrate
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 from the experimental measurements is shown below. The y-axis represent the fluorescence intensity at 520 nm. The excitation was done at 485 nm. The x-axis show the time for the measurements. The second set of graphs shown for each substrate represent the normalized values for the fluorescence intensity. This was analyzed because it represent the kinetic growth of the substrate proteins. As seen in the those graphs, the 50 % of max has been marked out as a dotted line. This makes it easy to interpret if the folding of the different substrate proteins was affected by the chaperone systems.
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 celsius. The bottom graphs show the normalized values from the top graphs.
Results for the EGFP-Aß1-42 substrate protein
Figure x. Results for EGFP-Aß1-42. Top graphs show fluorescene intensity divided by the start OD600 over 16 hours at 37 degrees celsius. The bottom graphs show the normalized values from the top graphs.
Results for the a-synuclein-EGFP substrate protein
Figure x. Results for a-synuclein-EGFP. Top graphs show fluorescene intensity divided by the start OD600 over 16 hours at 37 degrees celsius. The bottom graphs show the normalized values from the top graphs.
Results for the Tau0N4R-EGFP substrate protein
Figure x. Results for Tau0N4R-EGFP. Top graphs show fluorescene intensity divided by the start OD600 over 16 hours at 37 degrees celsius. The bottom graphs show the normalized values from the top graphs.
Results from the experimental measurements is shown below. The y-axis represent the fluorescence intensity at 520 nm. The excitation was done at 485 nm. The x-axis show the time for the measurements. The second set of graphs shown for each substrate represent the normalized values for the fluorescence intensity. This was analyzed because it represent the kinetic growth of the substrate proteins. As seen in the those graphs, the 50 % of max has been marked out as a dotted line. This makes it easy to interpret if the folding of the different substrate proteins was affected by the chaperone systems.
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 celsius. The bottom graphs show the normalized values from the top graphs.
Results for the EGFP-Aß1-42 substrate protein
Figure x. Results for EGFP-Aß1-42. Top graphs show fluorescene intensity divided by the start OD600 over 16 hours at 37 degrees celsius. The bottom graphs show the normalized values from the top graphs.
Results for the a-synuclein-EGFP substrate protein
Figure x. Results for a-synuclein-EGFP. Top graphs show fluorescene intensity divided by the start OD600 over 16 hours at 37 degrees celsius. The bottom graphs show the normalized values from the top graphs.
Results for the Tau0N4R-EGFP substrate protein
Figure x. Results for Tau0N4R-EGFP. Top graphs show fluorescene intensity divided by the start OD600 over 16 hours at 37 degrees celsius. The bottom graphs show the normalized values from the top graphs.
Figure x. Results for mNG-Aß1-42. Top graphs show fluorescene intensity divided by the start OD600 over 16 hours at 37 degrees celsius. The bottom graphs show the normalized values from the top graphs.
Results for the EGFP-Aß1-42 substrate protein
Figure x. Results for EGFP-Aß1-42. Top graphs show fluorescene intensity divided by the start OD600 over 16 hours at 37 degrees celsius. The bottom graphs show the normalized values from the top graphs.
Results for the a-synuclein-EGFP substrate protein
Figure x. Results for a-synuclein-EGFP. Top graphs show fluorescene intensity divided by the start OD600 over 16 hours at 37 degrees celsius. The bottom graphs show the normalized values from the top graphs.
Results for the Tau0N4R-EGFP substrate protein
Figure x. Results for Tau0N4R-EGFP. Top graphs show fluorescene intensity divided by the start OD600 over 16 hours at 37 degrees celsius. The bottom graphs show the normalized values from the top graphs.
Results for the a-synuclein-EGFP substrate protein