Start Date
2022
Description
Sec23 and Sec24 are Endoplasmic Reticulum budding proteins that assists various proteins in transport from the Endoplasmic Reticulum to the Golgi Apparatus. There are multiple isoforms of the Sec23/24 proteins, that serve the same function in cells.
We have evidence that Sec23/24 interacts with the glucose transporter GLUT1 in mammalian cells, and to confirm such interaction, we must able to purify and isolate Sec23/24 proteins expressed by mammalian cells. Therefore, our research goal is to clone mammalian expression vectors containing the DNA sequences of the Sec24a and Sec24b isoforms with the DNA sequences of affinity purification tags, such as a six-histidine (6xHis) tag and a hemagglutinin tag (HA), adjacent to the Sec24 DNA. Having previously cloned Sec23 expression vectors with affinity tags, successfully cloning Sec24 vectors will allow future study of both proteins. Once transfected into mammalian cells, these newly synthesized vectors will express the Sec24 protein with an affinity purification tag, which will allow us to purify each protein so that we can evaluate if they interact directly with GLUT1.
Recommended Citation
Bouwer, Seth; Griffen, Liz; and Arnoys, Eric, "Cloning Affinity Purification Tags into ER Budding Protein Sec24" (2022). Summer Research. 24.
https://digitalcommons.calvin.edu/summer_research/2022/Posters/24
Included in
Cloning Affinity Purification Tags into ER Budding Protein Sec24
Sec23 and Sec24 are Endoplasmic Reticulum budding proteins that assists various proteins in transport from the Endoplasmic Reticulum to the Golgi Apparatus. There are multiple isoforms of the Sec23/24 proteins, that serve the same function in cells.
We have evidence that Sec23/24 interacts with the glucose transporter GLUT1 in mammalian cells, and to confirm such interaction, we must able to purify and isolate Sec23/24 proteins expressed by mammalian cells. Therefore, our research goal is to clone mammalian expression vectors containing the DNA sequences of the Sec24a and Sec24b isoforms with the DNA sequences of affinity purification tags, such as a six-histidine (6xHis) tag and a hemagglutinin tag (HA), adjacent to the Sec24 DNA. Having previously cloned Sec23 expression vectors with affinity tags, successfully cloning Sec24 vectors will allow future study of both proteins. Once transfected into mammalian cells, these newly synthesized vectors will express the Sec24 protein with an affinity purification tag, which will allow us to purify each protein so that we can evaluate if they interact directly with GLUT1.