School of Science Division of Life Science 19 Functional Roles of Arfpr1 in Regulating Export of Cargo Proteins out of the Trans Golgi Network Supervisor: GUO Yusong / LIFS Student: WANG Xinnan / BCB Course: UROP 1000, Summer Apo-E is a protein that is involved in the metabolism of fats. It is encoded in humans by the gene APOE, and it has 3 kinds of alleles (E2, E3, E4). These alleles are different at one or two amino acids (E2 112C 158C; E3 112C 158R; E4 112R 158R), these differences alter APOE structure and function. For example, people with the homozygous APOE4 gene have a higher probability of developing Alzheimer’s disease. We only had plasmids containing the APOE4 gene in our lab. To get the plasmids with APOE3 and APOE2, we employed molecular cloning techniques to mutate the existing gene APOE4 into the other two alternative alleles. Cell Surface Receptor Interactome Related to Forgetting Supervisor: HIRANO Yukinori / LIFS Student: HAYASHIZAKI Yuki / BCB ZHONG Yuqing / BCB-IRE Course: UROP 1100, Spring Active forgetting is a critical yet poorly understood aspect of memory regulation. Building on Professor Hirano’s discovery of a gene-expression-based forgetting pathway, this project investigates the role of Kek2, a synaptic plasticity regulator, in dopamine-dependent memory modulation. To identify Kek2’s interactors, we constructed 18 expression plasmids encoding candidate proteins from prior proteomic data. Despite challenges in PCR amplification—including nonspecific amplification and template degradation—we successfully cloned Inx2, fax, CG17600, Kek5, and Spz5 into vectors for transfection in Drosophila S2 cells. Co-immunoprecipitation (co-IP) and Western blotting revealed limited Kek2-FLAG detection, prompting optimization of tag multiplicity and transfection conditions. Staining assays also failed to show expected colocalization between HA- and FLAG-tagged proteins, suggesting technical refinements are needed. Future work will employ touchdown PCR and nested PCR to resolve amplification issues, followed by systematic optimization of co-IP protocols to elucidate Kek2’s protein interaction network. This study advances understanding of active forgetting mechanisms and provides a methodological framework for probing membrane protein interactomes.
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