Expression and Purification of zebrafish GADD34 and CReP
Poster Number
05B
Format
Poster Presentation
Faculty Mentor Name
Douglas Weiser
Faculty Mentor Department
Biological Sciences
Abstract/Artist Statement
In eukaryotic cells, the endoplasmic reticulum (ER) regulates and processes a third of the proteins. Failure to maintain proteostasis, homeostasis of proteins, results in accumulation of unfolded proteins in the ER which is also termed as ER stress. In response to this, cells will activate unfolded protein response (UPR), triggering downstream events such as the attenuation of protein synthesis. CReP (Constitutive Repressor of eIF2α Phosphorylation) and GADD34 (Growth Arrest and DNA-Damage Inducible) are the two major regulators of protein phosphatase 1 (PP1) that helps regulate ER stress and inhibit UPR. Zebrafish are useful model organisms because they are easy to breed, maintain, and manipulate their genome. Preliminary data suggests GADD34 and CReP expression levels fluctuate with ER stress levels. However, there are currently no antibodies to detect zebrafish GADD34 and CReP. Multiple different constructs of GADD34 and CReP were expressed under different temperatures to find which construct is best expressed in BL21 cells, a strain of E. coli .
Location
DeRosa University Center, Ballroom
Start Date
28-4-2018 10:00 AM
End Date
28-4-2018 12:00 PM
Expression and Purification of zebrafish GADD34 and CReP
DeRosa University Center, Ballroom
In eukaryotic cells, the endoplasmic reticulum (ER) regulates and processes a third of the proteins. Failure to maintain proteostasis, homeostasis of proteins, results in accumulation of unfolded proteins in the ER which is also termed as ER stress. In response to this, cells will activate unfolded protein response (UPR), triggering downstream events such as the attenuation of protein synthesis. CReP (Constitutive Repressor of eIF2α Phosphorylation) and GADD34 (Growth Arrest and DNA-Damage Inducible) are the two major regulators of protein phosphatase 1 (PP1) that helps regulate ER stress and inhibit UPR. Zebrafish are useful model organisms because they are easy to breed, maintain, and manipulate their genome. Preliminary data suggests GADD34 and CReP expression levels fluctuate with ER stress levels. However, there are currently no antibodies to detect zebrafish GADD34 and CReP. Multiple different constructs of GADD34 and CReP were expressed under different temperatures to find which construct is best expressed in BL21 cells, a strain of E. coli .