School of Science Division of Life Science 16 Role of PRRC1 in Regulating COPII-mediated Export of Cargo Proteins out of the Endoplasmic Reticulum Supervisor: GUO, Yusong / LIFS Student: YAU, Yin Lam / BCB Course: UROP2100, Fall UROP3100, Spring UROP4100, Summer In the eukaryotic secretory system, endoplasmic reticulum (ER) and Golgi apparatus play important roles. During the secretion process, coat protein complex II (COPII) captures cargo proteins at ER and forms vesicles in order to transfer the proteins to Golgi apparatus. Previous studies have shown that PRRC1, a cytosolic factor, may be involved in COPII disassembly. Knockdown of PRRC1 can decrease the ER-to Golgi transport efficiency of EGFR and ShhN, and increase membrane associations of COPII coats. To investigate whether PRRC1 is critical for COPII-mediated ER export process, GST-PRRC1 was generated to carry out the in vitro vesicle formation assay. The results generally showed that PRRC1 could promote the cargo trafficking. Biochemical Characterization of Histone Variants and Post-translationally Modified Nucleosomes Supervisor: ISHIBASHI, Toyotaka / LIFS Student: HARIMOTO, Yuken / BCB Course: UROP1100, Fall UROP2100, Spring H2B is an essential histone protein, and studies about its functions can be done using modified DNA and observing the effects. In this UROP project, we confirmed the specificity of the primary antibody used to bind to the mutated H2B histones. HeLa cells transfected by plasmids are used as the source of mutant H2B proteins in this experiment. The confirmation is done using immunofluorescence staining and western blotting to ensure that it is specific in both scenarios. Through both methods, it was confirmed that the primary antibody specifically binds to the mutant H2B protein and shows a signal proportional to the amount of protein. Biochemical Characterization of Histone Variants and Post-translationally Modified Nucleosomes Supervisor: ISHIBASHI, Toyotaka / LIFS Student: SHEK, Wai Hei / BCB Course: UROP1100, Fall UROP2100, Spring RNA Polymerase is an enzyme that produces RNAs and is present in most species. Recently, demands to produce RNAs increased for research and therapeutic applications such as mRNA vaccine. In this study, we target to express recombinant thermophilic RNA polymerase. We use genomic DNA of thermophilic bacteria Thermotoga Maritima, and amplified both core subunit of RNA polymerase (rpoA, rpoB, rpoC and rpoZ) and sigma factor (rpoD) which is also necessary for the RNA synthesis by RNA polymerase. I could successfully amplified rpoA and rpoB, and rpoD. The rpoD fragment was cloned into bacterial expression vector pET29a, and the sigma factor protein expression is undergoing.
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