Graduation Year

2025

Document Type

Campus Only Senior Thesis

Degree Name

Bachelor of Arts

Department

Molecular Biology

Reader 1

Sarah Marzen

Reader 2

Erin Jones

Abstract

Firefly luciferase (Fluc) is a powerful bio-imaging tool that is well-suited for use in living organisms. The process of evolving Fluc mutants for selective processing of modified luciferins has compromised important properties of Fluc that aid imaging performance, including brightness. To restore and enhance these properties, we sought to screen mutants for improved brightness, and to identify specific residues that are important for this improvement. I constructed a library of mutations in residues 524-530 onto a prior successful mutant, P-TS-LGR. These positions were selected due to their proximity to the activity site during the light-emitting step of luciferase oxidation. Following screening, the brightest mutants were identified and quantitatively assessed for increase in luminescence. The best mutants have shown possible convergence at positions K524 and L526, indicating novel mutations that can increase brightness.

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

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