The aim of the project is to optimize the expression of the proteins amyloid-beta and tau in the bacteria Escherichia coli. These proteins are known to be linked to the development of Alzheimer's disease, though the cause of the disease is still unknown [1]. They are both found in the brain in and around our neurons, and during Alzheimer's disease they accumulate to form plaques and tangles [3, 4]. Because these proteins are prone to misfolding and aggregate they are hard to study in a laboratory environment, which has slowed down research. It’s here our project comes in, trying to facilitate the production of these proteins.

The chaperones we will study are GroEL, GroES, DnaK and Trigger factor. GroEL and GroES helps unfolded and misfolded proteins to fold correctly. DnaK helps with disaggregating and with unfolding of misfolded proteins. And Trigger factor helps the protein to remain unfolded during its synthesis.
To express the chaperones in the bacteria we will use a large plasmid containing all four chaperones and give them unique promotors, one for GroEL-GroES, one for DnaK and one for TF. The purpose of this was to easily be able to induce transcription of the specific chaperones in different combinations.

To measure our expression of these proteins we will fuse them with fluorescent proteins called E-GFP and mNeonGreen. Thereby we can measure the fluorescence to follow the expression of the fusion proteins. For every one of alzheimer - fluorescent fusion protein we will create a plasmid resulting in 4 combinations. These 4 plasmids will use a fourth promotor so that we can control this expression as well.