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You searched for +publisher:"University of North Carolina" +contributor:("Slade, Kristin M."). One record found.

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University of North Carolina

1. Slade, Kristin M. Protein diffusion in Escherichia coli.

Degree: Chemistry, 2009, University of North Carolina

This dissertation describes the creation of a system to provide insight about the affects of protein expression on intracellular diffusion. Fluorescence recovery after photobleaching (FRAP) is used to obtain diffusion coefficients. Nuclear magnetic resonance spectroscopy detection of 19F isotopic-enriched proteins and fluorescent labeling with FlAsH and were attemped before settling on FRAP studies with green fluorecent protein (GFP). To regulate protein levels in Escherichia coli, several vector and promoter combinations were tested. Eventually, a single vector was created containing the structural gene for GFP under the lac promoter and a test protein under the araBAD promoter. With this vector, the test protein was expressed at varying levels and GFP was expressed at a constant level. Although chymotrypsin inhibitor 2 was originally chosen as the test protein, it was quickly replaced by alpha-synuclein, maltose binding protein, tau-40, and calmodulin. My most important result is that regardless of the type or amount of protein that was co-expressed, the GFP diffusion coefficient remained constant. We conclude that expression of these soluble proteins has little to no effect on the diffusion of GFP. Several disadvantages of FRAP became apparent in the process of obtaining these data. FRAP and the other common techniques for measuring translational diffusion, present numerous obstacles when working with structures that are only slightly larger than optical resolution, such as Escherichia coli cells. To overcome these obstacles, I also developed a new method to assess diffusion using through-prism total internal reflection fluorescence microscopy with continuous photobleaching. Here, the theoretical basis for this technique is presented. I demonstrate its applicability by measuring the diffusion coefficient, 6.3 ± 1.1 µm2/sec, of GFP in Escherichia coli cells. Advisors/Committee Members: Slade, Kristin M., Pielak, Gary J., University of North Carolina at Chapel Hill.

Subjects/Keywords: College of Arts and Sciences; Department of Chemistry

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Slade, K. M. (2009). Protein diffusion in Escherichia coli. (Thesis). University of North Carolina. Retrieved from https://cdr.lib.unc.edu/record/uuid:6861cac7-4086-4538-ae91-b093a6c7699d

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16th Edition):

Slade, Kristin M. “Protein diffusion in Escherichia coli.” 2009. Thesis, University of North Carolina. Accessed January 26, 2021. https://cdr.lib.unc.edu/record/uuid:6861cac7-4086-4538-ae91-b093a6c7699d.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7th Edition):

Slade, Kristin M. “Protein diffusion in Escherichia coli.” 2009. Web. 26 Jan 2021.

Vancouver:

Slade KM. Protein diffusion in Escherichia coli. [Internet] [Thesis]. University of North Carolina; 2009. [cited 2021 Jan 26]. Available from: https://cdr.lib.unc.edu/record/uuid:6861cac7-4086-4538-ae91-b093a6c7699d.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Slade KM. Protein diffusion in Escherichia coli. [Thesis]. University of North Carolina; 2009. Available from: https://cdr.lib.unc.edu/record/uuid:6861cac7-4086-4538-ae91-b093a6c7699d

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

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