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Date of Award

4-27-2021

Document Type

Restricted Thesis: Campus only access

Degree Name

Bachelor of Science

Department

Biochemistry & Molecular Biol.

First Advisor

Dr. Laura Runyen-Janecky

Abstract

Sodalis glossinidius is a bacterial endosymbiont of the tsetse fly. It resides in various tissues, both intra- and extracellularly throughout the fly. Currently, the RJ lab grows Sodalis in vitro, using agar plates and liquid cultures. While these methods provide useful information about Sodalis growth and survival in certain conditions, it will be useful to have methods to understand the process of Sodalis invasion, survival, and replication within insect cells. Therefore, methods are presented here to measure Sodalis invasion of two insect cell lines, S2R+ and C6/36. Parameters for three assays, including an epifluorescent microscopy-based assay, a gentamicin protection assay, and a flow cytometry-based assay were established and used to measure Sodalis invasion of host cells. These methods will allow us to increase our understanding of how this endosymbiont, Sodalis, invades, survives, and grows within eukaryotic cells. Additionally, ysaV and orgA, which are components of type III secretion systems, were shown to not be required for Sodalis invasion of and replication within S2R+ cells, respectively. This suggests that Sodalis may infect C6/36 cells and S2R+ cells through different mechanisms, or that S2R+ cells readily endocytose Sodalis, independent of Sodalis’ type III secretion systems.

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