Title

Expression and Purification of zebrafish GADD34 and CReP

Poster Number

05B

Lead Author Major

Biological Sciences

Lead Author Status

Senior

Second Author Major

Biological Sciences

Second Author Status

Senior

Third Author Major

Biological Sciences

Third Author Status

Senior

Fourth Author Major

Biological Sciences

Fourth Author Status

Sophomore

Fifth Author Major

Biological Sciences

Fifth Author Status

Senior

Sixth Author Major

Biological Sciences

Sixth Author Status

Freshman

Additional Authors

Youngin Park, Senior, Biological Sciences

Jacob Scheel, Senior, Biological Sciences

Format

Poster Presentation

Faculty Mentor Name

Douglas Weiser

Faculty Mentor Email

dweiser@pacific.edu

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

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Apr 28th, 10:00 AM Apr 28th, 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 .