June 25th, 2008 ecoli
A while ago I posted a bit about my research woes. Well, I’m happy to report that my efforts since then have been fruitful. Creating the new constructs seem to have done the trick. There must have been something wrong with the original recombinant plasmid I was using (one that I didn’t actually create myself, by the way).
I’d never thought I would be so happy about seeing a little purple band appear on a little peice of paper before, but that band indicates the right protein is getting expressed and I can see it via western blot.
Posted in microbiology, my research | 10 Comments »
June 9th, 2008 ecoli
It’s been a while since I’ve blogged about my own work. I thought I’ve give an update.
I have the chaperone of the CS1 pathway cloned onto a vector and in a host strain. But for some reason, I haven’t been able to induce expression to high enough levels to be useful for an overlay assay, in the periplasm prep.
In english, Gram-negative Enterotoxigenic E. coli (ETEC) have two membranes. I strip the outher membrane off and collect the proteins in the periplasm space. But, for some reason, the protein I’m trying to collect, which chaperones other CS1 pilus proteins between the two membranes, is not to be found in the periplasm.
Therefore, I’m going to take the gene out of the plasmid vector its currently in and try putting it into a different one. Wish me luck.
Posted in microbiology, musings, my research | 1 Comment »
March 4th, 2008 ecoli
So there’s nothing quite like getting to the lab after a long week away (because of exams) only to find out all your frozen and refridgerated stuff had to be moved around to make room for the new grad students.
I only just got used to all my things being where they are, and it only took me 4 years!
I mean, I know I’m only a lowly undergrad, but I’ve been in this lab since before those newbs realized that you could major in biology.
I kid, though, the new grad students are good and I know them from their rotations in our lab. And my stuff is just on the very bottom shelf (does being an undergrad make me shorter?) so not a huge deal.
And since it seems that I got rejected from the research program I wanted to do at NYU, I’ll most likely be staying around for yet another summer on Long Island. It’s dissapointing but not all bad. I’ll get a lot of work done on my current project, which will come in handy for my senior thesis next year. Also, my PI said he’d pay me by the hour over the summer. Which, at $10 bucks an hour means I’ll be doing pretty well, in a lab I’m very familiar with, doing research I love, with people I like.
Also, not being in the city will mean much less distractions allowing me to study for the MCATs in July.
Posted in musings, my research | 2 Comments »
February 6th, 2008 ecoli
Enterotoxigenic E. coli is a scary bug that makes people sick. Immuno-naive people, tourists, children and the elderly in third would countries mostly.
I am working with the ‘alternate’ chaperone usher pathway of CS1 pili in ETEC. It’s known as the alternate pathway because its functionally similar but genetically distinct from the ‘classic’ PAP pathway that we all know and love.
The original thinking is that this is an example of convergent evolution, since there is no homology, as far as we can tell between the two systems. And, until now, it was thought that the subunit interactions ran on slightly different mechanisms.
My project is trying to show that the chaperone-usher interaction is like that of the PAP pathway, showing that this actually a case of ancient divergence rather than convergence.
If I can do that, it could help elucidate a new model of infection for ETEC. w00t.
I don’t have any significant results yet, but I’m presenting my proposal and preliminary work to my lab group on Friday, and am currently working on my presentation. I’ll probably upload it to this blog upon it’s completion. Expect to be kept updated on my project and its results.
ecoli out.
Posted in microbiology, my research | 2 Comments »
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