2 min read
It's been quite a busy summer with many developments outside of the lab that have given us a lot of new avenues to explore as we get back in the lab this fall.
First, I was invited by Scott to join him on the Juice Box podcast at Arden's Day to speak about our project. You can listen to the interview here.
I also spoke about the Open Insulin project at the Colorado State University Ethics Colloquium in September. You can watch the video here.
Meanwhile in the lab, we've continued working on getting the expression of the insulin gene in e. coli to work. In mid-summer, we were thinking that a small error in our sequence might have been preventing the e. coli from expressing the fusion protein, consisting of green fluorescent protein linked to human proinsulin. Our main centrifuge also hadn't been fixed yet, (but it is now!) so we were mainly looking into re-ordering the construct with the small error fixed.
As we talked over fixing the error and what we really needed and ideally would have in the construct, we determined we could resume lab work with the existing version, as the small error should not make such a significant difference that it would completely prevent expression; rather it should only add an extra residue to the proinsulin protein, which won't be relevant until we're past the purification stage. So we're reviewing and incorporating suggestions we've gotten from our own literature research as well as from various experts along the way, and revising our experiments accordingly. We're going to be looking into inducing the expression of the protein earlier in the e. coli growth trajectory, and using different concentrations of IPTG, the expression-inducing agent, as well as trying auto-inducing medium. We've already started to investigate the effects of changing these variables in last week's lab work.
To continue to grow our knowledge about insulin production, we've also started a literature review at our weekly meetings which will begin tonight. We'll be looking at highly-cited papers on insulin production to get a better sense of the details of how we should be proceeding. The first few of these, we're dusting off from the early days of the project to review what we already know, and then we'll be working to get broader and deeper knowledge to inform our efforts.
Finally, we'll be working to streamline our online infrastructure so that we can provide better updates with less effort across all the channels the project uses. Stay tuned for more updates!