Graduation Year

2025

Date of Submission

12-2024

Document Type

Campus Only Senior Thesis

Degree Name

Bachelor of Arts

Department

W.M. Keck Science Department

Second Department

Biochemistry

Reader 1

Bethany Caulkins

Reader 2

Kathleen Purvis Roberts

Terms of Use & License Information

Terms of Use for work posted in Scholarship@Claremont.

Abstract

Plasmodesmata are intercellular pores that connect plant cells together, establishing cytoplasmic and endomembrane continuities within the plant. This physical connection facilitates the movement of nutrients and other materials throughout the plant, as well as cell-to-cell communication. The novel protein Plasmodesmata 1 (PD1, 36 kDa) was extracted from the plasmodesmata of Arabidopsis Thaliana, and its sequence was determined, however the structure and function of this protein remain unknown. This research focuses on developing a purification procedure for PD1 to obtain pure protein for future nuclear magnetic resonance (NMR) structural analysis. PD1 protein was first expressed in e. coli, and bacterial growth was monitored using UV-Vis spectroscopy. A standard Ni-NTA column purification procedure was used to guide the development of a PD1 specific procedure, and several attempts at purification were performed to optimize the conditions for this protein. Successful purification occurred under denaturing conditions using Tris-HCl based buffers at pH 8.0 with all steps run at 4 °C. Next steps include an isotopically labeled expression and purification of PD1 so interrogation of the biomolecular structure of PD1 can be performed.

This thesis is restricted to the Claremont Colleges current faculty, students, and staff.

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