Closing Comments

This summer, I was fortunate to work with a team in Dr. Allison’s lab on a project characterizing importin binding to thyroid hormone receptor alpha. Importins are proteins that play important roles in transporting molecules into the nucleus of the cell; by binding to a target protein, and then binding to nuclear pore complex on the nuclear membrane, importins can translocate and release cargo across the nuclear membrane. Thyroid hormone receptor alpha (TRα) is a transcription factor; that is, it is a protein that regulates gene expression, in this case in response to the presence of thyroid hormone, or T3. The receptor shuttles back and forth between the nucleus, but the precise methods of localization of TRα have yet to be fully characterized.

[Read more…]

Protein Expression

In my previous entry, I mentioned that we would move on to actually expressing our protein constructs. The procedure for protein expression was filled with its own issues – finding the ideal conditions to get the bacteria to express our plasmid has been very slow-going, and involved a lot of trial and error.

[Read more…]

Sequencing and troubleshooting

Our goal this summer was to create fusion proteins of thyroid hormone receptor alpha with GST and His tags, which I described more in depth in my last entry. In order to do this, we had to insert the gene encoding TRα or its domains into an entry vector (plasmid), before moving on to later steps in the project.

[Read more…]

Protein tags

The project I worked on this summer is ongoing, so by the time I joined, a lot of work had already been completed. The first “phase” of our project involves creating a plasmid (or small, circular piece of DNA) that when expressed will yield our fusion protein (TRα or individual domains with a peptide tag fused to it).

[Read more…]

Abstract v. 2

My original project this summer was to focus on analyzing mitochondrial DNA of human remains. However, due to unanticipated difficulties in obtaining new samples for study, I was unable to work on that project this summer. Fortunately, I was able to work on another project in Dr. Allison’s lab this summer.

[Read more…]