Date Thesis Awarded

5-2021

Access Type

Honors Thesis -- Open Access

Degree Name

Bachelors of Science (BS)

Department

Chemistry

Advisor

John Poutsma

Committee Members

Rachel O'Brien

Kurt Williamson

Abstract

Bacteriophages represent a large portion of the biomatter on our planet, and many of them have yet to be fully characterized. Here we discuss the proteomic analysis of a particular Bacteriophage, Mycobacteriophage CrimD. This phage was discovered on the Campus of William & Mary and has had its genome characterized. We took the next logical step of proteomic analysis.

In our analyses we made use high pressure liquid chromatography paired with linear ion trap mass spectrometry to analyze the proteome of CrimD at specific time points after the infection of its host, Mycobacterium smegmatis. Additionally, we used nanospray ionization with in-house produced analytical columns and emitters to analyze our samples. These techniques had been previously used in our lab to analyze different bacteriophages but required significant optimization in order to successfully analyze CrimD.

In our analyses we found that we are able to see different proteins being expressed in the different time samples. Consequently, we were able to assign each time point to represent a different phase in the replication cycle of CrimD, namely the lysogenic and Early and Late lytic phase of replication. We were also able to assign many proteins with unknown function to specific time points, opening the door for further characterization of these proteins and CrimD.

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