Completion of more E. coli expression work, paving the way for algal transformation work.
Last week, we reached another milestone. We were able to successfully confirm that our modular expression system (derived from the pASapI plasmid) is indeed functional, and working well in E. coli. We used a green fluorescent protein as our observable marker and got some beautiful images. The results are promising because it means that our system is robust and should be capable of expressing other recombinant proteins (like insulin and oral vaccines) now that we have confirmed the genetic constructs are not only integrated, but are also expressing correctly so far.
Meanwhile, in Cork, Ireland, we were given a lab space in collaboration with IndieBio EU and the microbiology department at University College Cork. We proceeded to transform and begin testing each pASapI plasmid construct that Sebastian had assembled and sent to us.
Transformed E. coli harboring pASapI-aaDa (a spectinomycin resistance gene).
These DNA plasmids, which will be miniprepped from the cells above, will eventually be used for algal tranformations in the next coming weeks. We will be very busy preparing for Day 90 (IndieBio Demo Day), which will be live-streamed on YouTube to potential investors all around the world!
Insulin Update
We still expect to have some algal insulin expression work finished fairly soon. Our focus was pulled a bit by the roller coaster that is the startup life, but fear not, because the cell lysate produced from the E. coli containing the pASapI-proinsulin plasmid is waiting in a freezer to be probed with an anti-proinsulin antibody, which we are waiting on in the mail. Once we can confirm that the insulin is working in E. coli, we will officially have completed all of the objectives in the first half of the experimental work, and start completing the second half (which involves verifying that the insulin gene also works in our microalgae).
Onward, for science and human progress!
Antonio and Simon subculturing the microalgae strains in preparation for the algal transformation experiments.
A closeup of one of our happy wild-type algae strains (chlamydomonas) growing quite vigorously.
