At the End of the Summer

And here it is. The moment we have all been traveling towards: the end of summer research! I am actually rather sad that the summer is pretty much over already. It has been a festive summer with some great results!

This summer has seen some fantastic triumphs and some disappointing failures as far as research results go (which taught me valuable lessons such as make sure the stock cells are viable).To review, my research basically deals with modifying protein using unnatural amino acids to give them novel characteristics. As I had at least four projects I will attempt to clarify some of the projects (and also I enjoy lists). Most of the summer was spent in synthesizing the proteins that I needed, with later analysis and experimentation once that was successful.

PRMT1: The first project deals with a protein called PRMT1 which is a methyltransferase that is a part of the development of heart disease. Through the insertion of an unnatural amino acid caged group, I was able to control the activity of the enzyme through a light-activated switch. Much of the time spent on this project was to be able to properly synthesize it (as it has some temperamental conditions that it requires). After much trial and error we were able to successfully synthesize the caged-PRMT1 and now all that remains to do is to find an activity assay to analyze it with (which will be done in the Fall).

GFP-FY: The unnatural amino acid fluorotyrosine can be used to modify the pKa of GFP (a protein that fluoresces). Once again a number of trials were needed to successfully get green protein with the appropriate fluorotyrosine inserted. But once done, the fluorescence was able to be measured and we discovered some interesting trends of fluorescence. This will also be continued in the fall.

Glaser-Hay: The glaser-hay reactions (where two alkynes come together) have worked very successfully. We started letting it go overnight, but once we discovered that it degraded protein we went to a time-trial. This worked and produced a gorgeous trend and we are currently finishing up applications so it can be sent off to get a mass and final verification that it worked!

Microwave PCR: Doing PCR in the microwave is a really exciting development that would allow a 20-fold increase in the amount of product produced compared to typical thermal PCR. Or at least it would if it worked. Through much trial and even more error I have performed at least a dozen trials where we alter variables to try and get it to work. So far it hasn’t but we are still hopeful and later today I will try again.

The projects that I worked on were extremely exciting and interesting. I am looking forward to finishing up all of the projects and presenting them for my honors project.  This summer has further enhanced my love for research and I thank the Charles Center for this opportunity to do what I love. A happy rest of the summer to you all!

Comments

  1. sntrackenberg says:

    What type of work are you planning on doing when you continue your projects into the fall? Are you going to continue all of these projects, or narrow your focus to one or two of the projects? Even though i don’t work on this type of research, it is extremely interesting and I am interesting to see what your results are in the future as well as what you do to continue your research.