Applications of Computational Protein Design to Red Fluorescent Proteins

Author: Moore, Matthew Michaels

Year: 2013

Degree: Dissertation (Ph.D.)

Advisor: Mayo, Stephen L.

Committee Members: Hsieh-Wilson, Linda C.; Rees, Douglas C.; Arnold, Frances Hamilton; Mayo, Stephen L.

Option: Biochemistry and Molecular Biophysics

DOI: 10.7907/6876-A570

Abstract

The research comprising this thesis is presented in three chapters divided into two parts. Part One, Chapters 2 and 3, of the thesis focuses on the application of rational design and CPD to the core residues of FPs. Chapter 2 applies CPD to the well known red fluorescent protein mCherry. Design hypotheses in this work were driven by a desire to red-shift the fluorescence emission of the parent protein. Chapter 3 takes the most successful results from the mCherry system and attempts to applies these results to the far-red FP mPlum. These two proteins, mCherry and mPlum, share a directed evolution parent, mRFP1. Part Two, consisting of Chapter 4, presents the beginnings of a comprehensive study into the applications CPD for designing FP surfaces. The system used is based on DsRed, the oligomeric parent of monomer mCherry. Professional and personal acknowledgments conclude the thesis.

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