Team:UZurich/Parts
Parts
Working on two chassis simultaneously, using different versions of our parts for each organism was often a necessity. Especially Chlamydomonas reinhardtii, having a particularly high GC content, has a strong bias towards certain codons. This led to an increase in the number of parts used during the project as a lot of parts were codon optimized for C. reinhardtii.
Another issue facing us at the start of the project was the lack of iGEM projects about the plant immune system. Each of our receptors and all its modifications was new to the registry.
We therefore see our mission not just in pursuing a new biosensing system, but also in driving forward the development of standardized parts for plant immunity. We hope to have built a solid basis for future iGEM teams sharing our enthusiasm for plant PRRs.
Our constructs that were expressed in S. cerevisiae were assembled by Golden Gate Cloning and selected for on a synthetic dropout medium.
For C. reinhardtii we used Golden Gate Cloning to preassemble the constructs and then transferred them to a C. reinhardtii vector with In-Fusion cloning.
Basic Parts
The table below contains all our basic parts.
We created a lot of new coding parts for our project.
Many parts encode the cell surface receptors used for our biosensor while others are different versions of split-reporter genes which were fused to our receptors.
Clicking on the part name will take you straight to the part page on the registry!
Composite Parts
During our project we designed multiple composite parts. Most are for expression of our plant PRRs in different versions in S. cerevisiae or C. reinhardtii. These include the receptors fused to YFP, designed to visualize expression and localization in the cells and the receptor fused to a split-protein to assess the ligand-dependent dimerization.
Further there are composite parts containing split luciferases and proteins for dimerization. We used these to further characterize already existing parts and our improved versions.