Date Thesis Awarded

5-2021

Access Type

Honors Thesis -- Access Restricted On-Campus Only

Degree Name

Bachelors of Science (BS)

Department

Biology

Advisor

Margaret Saha

Committee Members

Kurt Williamson

Hannes Schniepp

Mark Forsyth

Abstract

The accessibility for the tools of traditional 3D printing have opened new avenues for constructing inexpensive, DIY bioprinters. This bioprinter was constructed by assembling a commercial, off the shelf 3D printer, and further modifying it to be capable of extruding living cells. The viability of E. coli and bacteriophage were tested, and the results demonstrate that both biological agents were capable of surviving the extrusion process. This bioprinter will be a useful tool for the additive manufacturing of cell laden bioinks.

Inexpensive, portable, and low equipment diagnostic techniques are ideal for field work where refrigeration and laboratory equipment are not always available. By utilizing the specificity of bacteriophage, a colorimetric-based biosensor can be created to identify the presence of specific bacteria in the soil. We developed a design for a mycobacteriophage biosensor in which phage are engineered to contain a horseradish peroxidase insert; after expression and release from the host, the enzyme can be detected through a colorimetric response after the addition of a secondary substrate. The mycobacteriophage were immobilized to a strip of nitrocellulose membrane treated with the cationic polymer polydiallyldimethylammonium chloride, and we determined that the phage remained stable and infective for at least 13 days post treatment.

Elucidating the interactions between host and phage during infection has biomedical and biotechnological relevance and can lead to the development of phage based tools. The results of the RNA-seq revealed that WC1 follows a standard lytic phage pattern of early, middle, and late phase gene expression. Only 16.6% (1,045/6,700) of the host genes were differentially expressed, with approximately the same number of upregulated genes as downregulated. A gene set enrichment analysis of both KEGG and GO host genes was performed, revealing an overall upregulation in translational process and stress and defense, and a suppression in energy metabolism and cellular macromolecule metabolism.

Available for download on Sunday, May 14, 2023

On-Campus Access Only

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