Senior Project Advisor
gene expression, ARO4 protein expression, RNA sequencing, Pollard lab
My capstone project sought to investigate the degree to which ARO4 gene expression is influenced by local vs distant genetic variants. This involved engineering allele-swap strains of ARO4 tagged with a co-expressing fluorescent protein. I then compared protein expression between each strain's genotype which provided an estimate of the proportion of the overall phenotypic variation that can be explained by genetic variation between the underlying genomes.
From my experiment I was able to conclude that ARO4 protein expression was complex and dynamic as both strain background and allele had genetic influences on ARO4 protein expression. In addition, not only did time change the total ARO4 protein expression, it also modifies the genetic differences. This was illustrated by how early in pheromone response there was a big difference in the background protein expression, while that difference became less pronounced as time went on. These results are consistent with the RNA sequencing experiment, which means that background can be attributed to RNA levels. The allele on the other hand was not consistent with the RNA sequencing experiment, which means the allelic effect can be attributed to protein synthesis or decay.
The goal of my capstone is to share what work I have done in the Pollard lab, as well as improve my capability to communicate scientific information to a general audience. To accomplish this, I will develop a webpage for the Pollard lab website that provides the process and results of our transformation optimization experiments. To be able to create allele swap strains for this experiment, I performed a yeast transformation. However, the Pollard lab’s transformation protocol needed to be optimized to improve transformation efficiency. I documented the process and results from my yeast transformation optimization experiments on a webpage for the Pollard lab website, with the intention that it would be a resource for other research laboratories or prospective students who are interested in learning more about the Pollard research lab.
Tietz, Nadine, "Cupid’s Arrow: A Tale of a Complex and Dynamic Protein" (2022). WWU Honors College Senior Projects. 537.
Subjects - Topical (LCSH)
Gene expression; Nucleotide sequence; Saccharomyces cerevisiae
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