Summer cell cycle research

A summer of protein overexpression and harvesting awaits me!  I will be trekking into the Integrated Science Center bright and early every day to acquire purified truncations of the cell cycle protein Ulp1.  This will involve cloning the DNA coding for these truncated proteins into circular expression plasmids with an affinity tag, and transforming these cloned plasmids into bacteria.  Then, I will chemically compel these bacteria to produce buckets of the protein product.  Following affinity purification, I will use these purified truncations of Ulp1 protein to assay their ability to bind to the yeast Small Ubiquitin-like Modifier (SUMO), in hopes of isolating what portion of the protein is responsible for SUMO binding.  This will be accomplished, hopefully, through the use of another affinity column with SUMO immobilized on beads.

Why do all this?  Well, SUMO is a nifty little protein which is tacked onto other proteins to modify the localization, activity, or function of the target protein.  Ulp1 is a SUMO protease, which means it cuts SUMO off of sumoylated proteins.  A better understanding of how proteins become sumoylated or desumoylated has the potential to shed light on a plethora of cellular processes on the molecular level.  So, I embark to find what portion of Ulp1 is necessary to target it to SUMO, and ultimately sumoylated proteins.

My experiences with molecular cloning this semester has been humbling; although the techniques for doing it have been honed over the entire lifespan of modern molecular biology as a discipline, it still has the capacity to frustrate even the most meticulous, experienced, and patient researchers.  That being said, I think with my attention undivided and focused at the task in hand, I will be able to surmount any obstacles that molecular bench work puts before me.

Good luck to everyone else doing summer research!