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It is important for proteins with unknown functions that we determine characteristics about them to better understand their cellular role. Throughout this investigation, our aim was to determine the function of the known protein 2O14 in bacteria cells. The bacteria cells were transformed, inoculated, and finally isolated in a bacterial pellet. Protein induction followed this step and after this our protein of interest, 2O14, was purified from the bacteria cells using Ni-NTA affinity chromatography. Upon purification, we performed an SDS-PAGE analysis, in order to determine where the protein 2O14 was present in our purification steps, a Bradford Assay was then performed in order to determine the concentration of 2O14 in the cells. From these tests, we were able to determine that the protein was present in the first elution of the purification process. After creating a standard curve, our concentration of 2O14 was found to be approximately 7.27 mg/mL. Through further investigation, using p-nitrophenyl acetate as a substrate, we will be able to determine the activity and the kinetics of our protein 2O14 in which we hope to use to find its overall cellular role.
Johnson, Colton; Sprankle, Ken; and McCoun, Chris, "Determination of the Structure and Function of Protein 2O14" (2021). Biochemistry and Molecular Biology Presentations. 16.
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