Mutagenesis of Dbp5-VC for screening of Nab2 association

Publication Date

2026

College

College of Sciences & Mathematics

Department

Biology, Department of

Student Level

Undergraduate

Faculty Mentor

Rebecca Adams, PhD

Presentation Type

Poster

Summary

Saccharomyces cerevisiae is a budding yeast species used in biological molecular research to study eukaryotic cell processes. The process of mRNA export out of the nucleus is unique to eukaryotic cells due to the presence of a nucleus and the necessity of translation to proteins in a different part of the cell. When mRNA is transcribed in the nucleus, several associated proteins bind, including Nab2 and Mex67, which are the two removed by Dbp5 at the nuclear envelope. This removal of Nab2 and Mex67 but not any other associated protein raises the question of how Dbp5 selects for these proteins specifically. It can be hypothesized that Dbp5 and Nab2 bind at one specific and unique site. To test this, a mutant library of Dbp5 will be created and then tested to see if Nab2 still binds with the randomly mutated Dbp5. A conclusion this research has found is the concentration of manganese needed in order to induce a significant amount of mutations as well as have sufficient DNA to continue with the experiment. Additionally, a successful Gibson Assembly experiment has been completed but yielded a smaller amount of colonies than expected. Future directions would be to create more colonies to make a larger library of mutant plasmids. To do this, gel extraction has been a technique used to try to have more clean Dbp5.

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