I spent the first six weeks of my summer break staying on campus to intern with Dr.
Erika Sorensen-Kamakian. Two of my classmates, Nate Powell ’27 and Khush Ochilov ’28, were also on the job with me. Our research focused on the effects of loss-of-function (LOF) and gain-of-function (GOF) mutations in the glp-1 gene in C. elegans, a small roundworm. The LOF mutants were sterile and could not reproduce, but they had an increased lifespan due to improved proteostasis, or protein maintenance. The GOF mutants maintained their germline at an extreme level, leading to tumor formation.
Most of the research focused on two different ways that proteostasis could be
improved: through an improved proteasome or through improved proteases. The
proteasome is a protein-degrading complex made up of multiple subunits. To image the proteasome, we grew about 10 million wild-type worms, which underwent lysis and
centrifugation at 100,000 × g to isolate the proteasome. Using anion-exchange
chromatography, we determined the concentrations needed to calculate the amount of
proteasome required for BCA and CTL assays. Some of the proteasome sample was also sent for imaging.
Protease experiments were another major part of our research this summer. The
four protease-encoding genes we focused on were tpp-2, sch-1, lonp-2, and K12H7.4.
Using PCR, we amplified these genes. After running a gel to confirm PCR success, the gel extractions were used in Gibson cloning. The resulting products were then plated and screened. Successful minipreps were sent for sequencing, and confirmation of the correct sequence could lead to metal-affinity chromatography to isolate the protein.
Lastly, I would like to thank both Dr. Erika Sorensen-Kamakian and Dr. Walter Novak for helping guide me through this internship. As I had never worked in a lab before, I had a lot of questions for them, but their technical and biological knowledge was a critical resource throughout the summer. I greatly enjoyed working with them and learning from their expertise.
