The Roux Lab Summary

The Roux Lab is one of the labs at Sanford Research that focuses on proteins. The goal of the lab is to map the proteins of the nuclear envelope using BioID. This may sound like a simple goal but figuring out where each protein is located but it is a daunting task. With the lab trying to map the proteins, it is necessary to develop efficient ways of seeing how proteins interact. In the past years of the Roux lab, they have developed a system to see which proteins interact with other proteins, through a slightly mutated protein, BioID (BirA). The way that BioID works is by attaching to a protein of interest. Then with the help of ATP within the cell and excess Biotin that was introduced into the cells, the lab is able to use anti-bodies to see which proteins were biotinalated. Over the past few years they have been able to create BioID2 which is a smaller more efficient biotinalator. In this method all of the proteins that interact get biotinalated, but there is not a method yet to see if just two proteins interact with each other.
With the help of Dr.
Roux, Daein, and Gaya, this summer I will help develop a way to split the BioID enzyme into two parts and then attach the parts onto two proteins of interest. When the two separate proteins then interact with each other, the BioID molecule will be complete and have the ability to biotinalate. Knowing that Jun and Fos interact with each other within the nuclease, we have attached parts of BioID to it. As of yesterday, we have confirmed that it is possible to split BioID and that it will still biotinalate, and now for the rest of my research I will be continuing to test different arrangements of BioID and BioID2. The testing will include repeated cloning, immunofluorescence, and western blotting. The purpose of cloning is to change the type of BioID inside of the cell, and the immunofluorescence and western blot will show us the
results. By the end of the summer we expect that we will have at least 4 different ways to configure BioID on the proteins. These will be available to other labs to buy. In the Roux lab, a big focus is on helping other people’s research. Our lab right now is in transition, with two people leaving within a few weeks. When I started there was the PI, Dr. Kyle Roux, two post docs, Kyle Noble and Daein Kim, three research associates, one graduate student, and one research assistant. Two weeks ago Daein left to go work in Texas and next Wednesday Ben will transition to work in the PROMISE lab. With 3 summer interns there is a variety of lab members. Daein was a lab coordinator until he left, so currently Dr. Roux is the acting lab coordinator. Out of the lab members I typically work under Gaya and she is very informed on my project. Since there are only three summer students, I feel like I fit in the lab but have the ability to ask a number of people if I have any questions. I have been invited and included in all of the activities of the lab, including a farewell lunch and a happy hour (water for me). After journal clubs and lectures my menotors usually do not follow-up with me but if I have any questions they are willing to answer it to the best of their ability.