Luke Cline

Generating a reliable reporter for SSA4 mRNA export during heat shock

For gene expression to occur in a cell, the DNA containing the genes must first be transcribed into mRNA, the mRNA must be exported from the nucleus to the cytoplasm, and the mRNA in the cytoplasm must be translated into protein. Thus, an essential process of eukaryotic cells, such as S. cerevisiae, is mRNA export. The export of mRNA can be regulated to change gene expression in response to changing conditions to keep a cell functioning. This project centers around a group of mRNAs selectively exported during heat shock in yeast cells, specifically the heat shock mRNA called SSA4. We seek to understand why SSA4 mRNA gets exported while most mRNA transcripts remain trapped in the nucleus.

Answering this question requires a reliable way to accurately assess the export of SSA4 mRNA in the yeast cell. My project is to generate new reporter plasmids with the SSA4 gene that accurately reflect this gene as it is found in normal yeast cells. Generating these more accurate plasmids will remove the region called U1A that we hypothesize to be hindering the functionality of our reporter plasmids.

From our experimentation with the new reporter plasmids, we garnered inconclusive results. We can tell that new reporter plasmids have been successfully generated, but no definitive conclusions can be made in the steps beyond plasmid generation. In future experiments, a time course with the plasmids generated here should be conducted

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