The amoeba Dictyostelium discoideum is a single-celled organism that has shown great promise in terms of research, especially in the modeling of cancer cells because of their responses to starvation. Our objective is to utilize these similarities to study the interaction between a protein kinase, TklA and a phospholipase, PldB, for the regulation of the actin cytoskeleton in Dictyostelium discoideum. Although previous studies have studied other genes and their impact, our pair of genes have not been studied together in-depth. We tested the amount of cells that would adhere to the substrate over a period of 40-minutes after starvation and studied the upstream or downstream effect of the mutations, both on the protein kinase TklA and the phospholipase PldB, by comparing the two mutants and their double mutant.
We measured cell concentrations before starvation and then at set intervals for a total of 40-minutes. The differences between the initial and interval measurements were used to calculate the percentage of cells that remained adhered to the flask, which acted as the substrate.
In the end, through the analysis of the two mutants and the double mutant, the connection between the two mutants was established that the phospholipid gene, PldB, acts downstream to gene of the protein kinase, TklA, Our results were justified as significant through our usage of the statistical student t-test as the cut-off for separate populations was p<0.05.