Utilizing β-gal to visualize selective transport of the SSA4 gene in S.Cerevisiae

Authors

Ian M. Staertow, Belmont UniversityFollow
Rebecca L. Adams PhD, Belmont UniversityFollow

Publication Date

2025

College

College of Sciences & Mathematics

Department

Biology, Department of

SURS Faculty Advisor

Dr. Rebecca Adams

Presentation Type

Poster Presentation

Abstract

In eukaryotic cells such as Saccharomyces cerevisiae, protein synthesis begins with the transport of mRNA from the nucleus to the cytoplasm. In order for mRNA to leave the nucleus, it must first bind to specific transport proteins which can then facilitate transport via the nuclear pore complex (NPC), a selectively permeable membrane within the cell's nuclear envelope. Under environmental stress, such as heat shock, S. cerevisiae cells modulate transport rates. In this case, in heat stress or “heat shock” most mRNA transcripts will remain in the nucleus. The SSA4 transcript encodes for a chaperone protein which facilitates the subsequent export of this otherwise “stuck” material. This then raises the question at hand, how does this transcript go about exporting mRNA from the nucleus under these conditions, while other transcripts do not? To investigate this mechanism overall, my project focuses on developing a tangible, phenotypic reporter system for selective SSA4 transport under a select set of conditions. Furthermore, I have inserted a bacterial plasmid containing the β-gal gene into the SSA4 locus in wild type S. cerevisiae. With this integration, we expect to see the production of a visible blue expression in the presence of the X-gal substrate within our S. cerevisiae cultures following heat stress or “heat shock”. The β-gal gene was inserted to the SSA4 gene locus using CRISPR-Cas9 technology and was inserted into a wild type strain of S. cerevisiae (yeast), we later confirmed this via. a series of PCR analysis. These repeatable methods can be used in future implications by utilizing modified S.cerevisiae strains that will reflect identifiable proteins involved in this molecular mechanism, among others.

Recommended Citation

Staertow, Ian M. and Adams, Rebecca L. PhD, "Utilizing β-gal to visualize selective transport of the SSA4 gene in S.Cerevisiae" (2025). Science University Research Symposium (SURS). 278.
https://repository.belmont.edu/surs/278