My project this summer is on the divisome, which is the structure responsible for bacterial fission. Specifically, it's on FtsA and how it interacts with FtsZ.
Fts stands for Filamentous Temperature Sensitive, and describes how when heated these proteins form filaments! [EDIT: No they don't, see later to find out why!] More relevently, FtsZ polymerises to form a massive ring, which contracts via a GTP/GDP dependent mechanism, bringing the membrane with it, until it draws tight like the drawstring of a sleeping bag, pinching off two distinct bacteria. FtsA is a membrane-associated protein that mediates division by binding to FtsZ, but is not essential for this process.
So, technical sciency part over; what has my first day been like? Well after realising cheese and pitta bread sandwiches for breakfast was a mistake, I arrived at the lab where I met Dr. D and was filled in on the basics of what I'll be doing, at least for a few weeks. Then I was introduced to the post-grads who all seemed pretty nice people, as well as the technician who was given permission to shout at me. The ground rules for the lab are:
- No open-toed sandals in the lab
- No food in the lab
- No drinking the technician's coffee, ever
- Keep the lab tidy
- No lab coats in the research office
After smuggling my lab coat out of the research office I entered the lab, where Dr. Mohammed Jamshad (also a really nice guy, and my de facto mentor for this project) showed me around. He's currently harvesting ZipA proteins from E. coli, which are acquired in a similar manner to the FtsA I shall be harvesting later, as well as sharing similar roles (possibly).
So first we removed the growth media from the autoclave (a sort of pressure cooker used to sterilise things) and added ampicillin to kill any unwanted microorganisms that might sneak in when our backs were turned. To this we added 1% volume of a culture of E. coli, transformed with a plasmid containing ZipA and ampicillin resistance. Once this was done, we stuck them in an incubator and swirled them for about 3 hours at 37 degrees celcius and at 200rpm, securing the flasks with wads of tissue stuck between the vessel and the holding ring. That thing looks like it'd be unpleasant to clean, so I'm definitely going to remember to do that.
Then we cleaned up the glassware. In order to clean the culture flask Dr. J used a pink chemical which was presumably some sort of bleach, adding some to water and then swirling it in the flask, before leaving it for 30 minutes.
Now, it is here that I noticed a lab-based hazard. If there's anything I have learnt from organic chemistry practicals, it is that very dangerous chemicals tend to look highly edible (my substituted phenyls tend to look like gingerbread, although this is probably down to appalling technique). Moreover, laboratories have a tendency to make me very hungry (again, probably due to lab sessions traditionally ending at 12, lunchtime). Finally, this bleachy stuff looked and smelled of sherbert.
I voiced my concerns to Dr. J, who reassured me that it did indeed look and smell like sherbert, but was also very bleachy, thus I should refrain from sampling it. I was learning more and more about lab behaviour and scientific technique by the minute.
At the moment I am on a short break, in a while I shall be putting together some IPTG to induce translation of ZipA, as well as checking the flasks to make sure they're not on fire, and that the optical density is about 0.6-0.8 (which I thought was surprisingly high).
First day is going very well indeed. :D