Date of Award

Summer 2025

Rights

Access is available to all users

Document Type

Thesis

Degree Name

Master of Science (MS) in Biology

Department

Biology

Abstract

Acne vulgaris is one of the most common skin diseases globally. One key aspect of acne pathogenesis is the disturbance of healthy strain diversity of the skin bacterium Cutibacterium acnes. Many acne treatments are antimicrobial; however, these treatments are generally nonspecific and carry a variety of side effects in addition to contributing to rising antibiotic resistance. Lytic bacteriophages (or phages) are viruses that infect and kill bacteria. Their lytic lifestyle is facilitated by enzymes called endolysins. Phages have been studied as an alternative to traditional antimicrobials in acne treatment and have shown reduced C. acnes load in acne lesions and improved symptoms in patients. However, most current research uses intact phages. This provides logistical challenges as the phages must be cultured, maintained, and kept viable until treatment. Here, we sought to recombinantly produce and extracellularly secrete the P100.1 phage endolysin in a Lactococcus lactis production system. However, throughout this process, we encountered several critical roadblocks and were ultimately unable to achieve recombinant production. We attempted production in three expression plasmids, one of which was chimerically produced. In the absence of results testing the antimicrobial effects of the recombinant endolysin, here we will discuss and investigate the challenges and impediments we faced during this project. We will discuss attempts with various cloning techniques, chimeric vector design and production, and the exhaustive efforts undertaken in our attempt to achieve production.

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