<p>In our research, we have undertaken the construction of a plasmid to serve as a pivotal tool in our experimental endeavors. This plasmid has been meticulously designed to facilitate the expression of the CAHS3 protein, which is tagged with the V5 epitope.</p>
<p>Designed was a vector for the expression of CAHS3 protein tagged with V5 in the experiment. In this process, the XbaI and NotI enzymes were chosen to facilitate the insertion of the PCR-amplified product into the pUAS-attB vector. The use of agarose gel electrophoresis confirmed the successful amplification of CAHS3 cDNA via PCR.</p>
<p>Before generating transgenic flies, immunofluorescence and western blot assays were conducted to validate the expression of CAHS3 in cultured cells. The original construct obtained from Addgene allowed successful expression of CAHS3 protein in mammalian cells. Subsequently, the newly created plasmid was transfected into Drosophila S2 cells. Immunofluorescence staining revealed the successful expression of CAHS3, and western blot analysis confirmed the presence of CAHS3 protein with the correct molecular weight. Tubulin was employed as a loading control.</p>
<p>Following the generation of flies with the specified genotypes, another western blot analysis was performed on the flies. The results clearly demonstrated that CAHS3 was successfully expressed in these flies.</p>
<h3>Modeling</h3>
<p>We also built a mathematical model to help us better calculate and observe the results of the experiment.</p>
<p>In the model, we analyzed the imported testing images through the RGB color system and then analyzed the obtained data with the T-statistics. Using the computer program we developed, the color oscillation of the imported pictures of the fly eyes can help to identify the arrangement patterns of the compound eyes. The program then outputs with data that could be used to do statistics to determine whether there is morphological difference between different experimental groups.</p>
