Team:Linkoping Sweden/construct
Our constructs
Chaperone systems
We designed one superduper-plasmid with the genes for all three cheperone systems we are useing, GroEL/GroES, DnaK and Trigger Factor.
We put in different promotors for these genes to be able to regulate the expression. We used the the iGEM backbone psb1C3 for our superplasmid. The psb1C3 backbone contains a gene for chloramphenicol resistence.
For the GroEL/GroEs chaperone system we used the T7 promotor which is induced by addition of IPTG.
For the Trigger Factor gene, we chose a tetracycline promotor, therefor we also had to put in the gene for tetracycline resistance.
The tetracycline promotor is induced by adding tetracycline. The last promotor we used in the superduper-plasmid was a rhamnose promotor called pRha, this promotor was used for the DnaK gene and is induced by adding L-rhamnose.
Amyloid-beta
We designed one plasmids with amyloid-beta bound to EGFP with C-terminus and one plasmid with Amyloid-beta bound to mNeonGreen protein with C-terminus. For these plasmids we used an arabinose promotor called AraC which is induced by addition of arabinos. We put these substrates in the psb1A3 backbone from iGEM. The psb1A3 backbone has a gene for ampicillin resistance.
Tau
We designed one plasmid with Tau bound to GFP with N-terminus and one plasmid with Tau bound to mNeonGreen protein with N-terminus . We used the arabinose promotor and the psb1A3 backbone here as well.
