Belmont University Research Symposium (BURS)

Publication Date

Spring 3-31-2022

College

Sciences and Mathematics, College of

Department

Biology, Department of

BURS Faculty Advisor

Rebecca Adams, PhD

Presentation Type

Poster Presentation

Abstract

In eukaryotes, mature mRNA is transported out of the nucleus, the site where it is generated, to the cytoplasm, where it is translated into protein, and through the nuclear pore complex, which are doorways in the nuclear envelope. A host of proteins that regulate proper mRNA export have been identified, but the process is still not fully understood. A gene called BOP3 has been identified as having similar functionality as proteins that aid in mRNA export, but very little is understood about this gene. In order to uncover its function, a synthetic lethal screen was performed in the Adams lab by a previous student, leading to a strain that is mutated in an unknown, functionally-related gene, which we call bop3synthetic lethal gene 1, BSL1. The bop3D bsl1 mutant strain has been transformed from a library of 2m plasmids to identify those that have gained the ability to lose the BOP3 plasmid that allowed viability. The goal of this project is to isolate the plasmids from this screen. Our hypothesis is that identification of the rescuing plasmid will enable identification of BSL1 and thus inference of Bop3 function. In the process of this project, we confirmed the bop3Dbsl1 strain by preforming colony PCR and gel electrophoresis. Thus far, rescuing strains that demonstrated presence of a 2m plasmid and loss of BOP3 were then cultured, mini prepped, and transformed into bacteria to isolate rescuing plasmid. We will next sequence the plasmids and confirm their rescue of the bop3Dbsl1 strain. Only after identification of BSL1, can the function of BOP3 and its role in mRNA export be better understood. This research aims to increase our understanding of the proteins involved and overall process of successful mRNA export.

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