Date of Award

6-4-2014

Document Type

Thesis

Publisher

Santa Clara University

First Advisor

Lou Hom

Abstract

Bioluminescence is the ability of naturally occurring organisms to emit light. Many instances of bioluminescence have been identified ranging from fireflies to bacteria. The protein responsible for bioluminescence in all of these organisms is luciferase. The exact function and structure of this protein are still unknown to this day. This project aims at better understanding luciferase as well as enhancing its bioluminescent properties. In order to work with luciferase, we had to isolate all of the genes necessary for bioluminescence. The six genes necessary for bioluminescence are Lux A,B,C,D,E,G. We isolated each of these genes from original bacterial DNA. We then put these genes back together into our own synthesized plasmids through a method called Gibson Assembly. These plasmids were then mutated through a process called site-directed mutagenesis. The mutated plasmids were introduced to E.coli cells and mutated protein were produced and screened for enhanced bioluminescent properties. We were able to successfully isolate all of the DNA fragments necessary for bioluminescence. We were then able to successfully create one of the two plasmids necessary for bioluminescence. The successful creation of this plasmid shows that we were able to successfully isolate and combine the DNA fragments. By shifting a few small experimental parameters we should be able to complete our second plasmid and achieve bioluminescence, and move onto mutating the DNA. Luciferase doesn’t have many applications in today’s world because it remains so understudied. By mutating luciferase, we hope to shed some light on this protein and allow it to be widely used. One possible application is being explored by a company called the Glowing Plant Project, located in San Francisco. They are using the luciferase and inserting it into plant genomes to create glow in the dark plants. These have some amazing promise not just aesthetically in our gardens but also as a potential light source for third world and developing countries.

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