Team:UZurich/Progress Reports
On this page we will post updates on our project throughout the summer and fall.
If you don't know what our project is or what we are doing, it is best to first read our project description first!
Progress report 19th of July
After familiarizing ourselves with the lab equipment and numerous techniques, we are currently in the middle of our in vivo analysis. We designed a plasmid with the lacI system, antibiotic resistance against streptomycin/spectinomycin and a reporter gene (yellow fluorescent protein). A multiple cutting site downstream of the latter enables us to insert our gene of interest which is then linked to the YFP.
With the help of BLAST and this paper, we were able to select 11 genes that are likely involved in the formation
of the phage compartment that we want to artificially induce.
After amplifying these genes with PCR, we ligated each of them into one of the described plasmids. As soon as we had
ensured that the genes had been successfully been inserted into our plasmids, we transformed 11 Pseudomonas chlororaphis
cultures with the 11 plasmids we obtained, of course, one plasmid per culture. We are working on getting
the transformations done correctly. It is a bit difficult, since we need to use electroporation and it is less efficient
than usual transformation with for example E. coli. Before August we will be able to start with imaging.
For the imaging we will first infect all cultures of P. chlororaphis with our phage. Then the phage compartment will
naturally form, but additionally the bacteria have a plasmid carrying a gene linked to YFP. So we are looking closely at bacteria
that are expressing our gene of interest linked to YFP AND also have been infected by the phage (and therefore are developing the compartment).
In these bacteria we can evaluate the role of that gene in forming the compartment.
The involvement of the proteins will be assessed by their location. Our hypothesis is that if the protein is not
imperative for the phage compartment it will be spread evenly throughout the cell, whereas the fluorophores linked
to essential proteins will be bundled in a circular structure.
This hypothesis is based on the fact that gp105, the main component of the compartment is first distributed evenly
in P. chlororaphis cells and upon infection it is concentrated at the compartment.
Beside our lab work, numerous milestones regarding human practices (an iGEM medal requirement) have already
been accomplished. Firstly, we taught high school students about synthetic biology, our project and iGEM through
holding presentations at various high schools. Furthermore, two of the high school students will be performing
experiments crucial to our project as their end-of-school project (Maturarbeit). We are looking forward not only to
teaching them essential synthetic biology techniques, but we also want to encourage them to think critically about how
fundamental research is done in academia today. For learning more about the ethical implications of synthetic biology
we met with Anna Deplazes, an ethics lecturer at the University of Zurich.
Also important, we have the first version of our wiki online!
- check it out here: https://2019.igem.org/Team:UZurich
Much is to be done, but we managed to give you a flavour for the content that is going to be released throughout the project.
The next progress report will be released after the imaging and analysis for the first 11 candidate genes has been done.
By then the version 2 of our wiki will be released, that version will completely update the layout.