Graduation Year

2018

Graduation Month

May

Document Type

Thesis - Campus Access Only

Degree Name

Bachelor of Science

School or Department

Biochemistry and Biophysics Program

Major

Biochemistry

Faculty Mentor Department

Biological Sciences, Division of

Faculty Mentor

Scott Samuels

Subject Categories

Pathogenic Microbiology

Abstract

The bacterium Borrelia burgdorferi is the causative agent of Lyme disease, the most common arthropod-borne disease in the Northern Hemisphere. This ailment affects over 300,000 people in the United States annually, and can cause symptoms such as fever, joint stiffness, fatigue, carditis, and neurological issues. One of the most fundamental processes in any cell, including B. burgdorferi, is the transcription of DNA into an RNA messenger, which is then translated into proteins. Transcription is carried out by a multisubunit molecular complex called RNA polymerase, which is recruited onto the DNA strand and synthesizes RNA by reading the DNA. RpoD, also known as σ70, is the subunit of RNA polymerase in bacteria that recognizes the RNA polymerase binding site on DNA. In all bacteria studied to date, the size of RpoD is 70 kilodaltons (kDa). Although the rpoD gene in B. burgdorferi appears to encode a 70-kDa subunit, two lines of evidence suggest that only a 50-kDa protein is produced in these bacteria. In order to biochemically dissect the function of this truncated RpoD in B. burgdorferi, I have overexpressed and purified it to near homogeneity using recombinant DNA methodologies. The putative RpoD protein was then assayed for activity using an in vitro transcription assay. Future directions for this project would include obtaining a structure of the truncated RpoD from B. burgdorferi and analyzing its regulation.

Honors College Research Project

Yes

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© Copyright 2018 Bethany Crouse