Difference between revisions of "Team:Linkoping Sweden/Description"

Our Workplan

We designed a superduper-plasmid with the code for three different chaperones, GroES/GroEL, DnaK and Trigger factor each with a different promotor. The purpose of this was to be able to induce transcription of the specific chaperones in different combinations. Four other plasmids were designed for the substrates; E-GFP bound to Amyloid-beta or Tau and M-Neongreen protein bound to Amyloid-beta or Tau, for these substrates we used a fourth promotor. After some research, we found that both E-GFP and M-Neongreen are suitable fusion proteins for proteins that are hard to express in E.coli and decided to use both.

The plasmid with chaperones and the plasmid with substrates have different antibiotic resistance genes to make it possible to select the bacteria’s that expresses both plasmids.

We will induce the expression of the chaperones before we induce the expression of the substrates to make sure that there are chaperones present when the translation of the substrates begins.

Both fusion proteins are fluorescent which will enable us to get an indication of the expression. Further we will use several methods to detect and quantify our proteins.

The overview of the project is summarized in figure 1 below. As can be seen in the figure, chaperones and conditions will be tested with various experiments, with the purpose to find the best setup for further detailed studies. The detailed studies will involve modeling with systems biology, where the modeling team will give suggestions on what the laboratory team shall do to achieve the desired task. The flow of data between the laboratory and the modeling team will eventually result in the final data, with is the optimized setup for the expression of our peptides.