Student Reflection - Stacy Batres

Identifying B56α and B56ε specificity

This summer, with the help of the EURECA FAST Award, I had the privilege of working under Dr. Joni Seeling on an ongoing research project that focuses on the Wnt signaling pathway. The Wnt signaling pathway is important to several developmental processes that cause the formation of cancer cells if mutated or unregulated. Within this pathway there is a regulatory complex called Protein Phosphatase 2A (PP2A) which is composed of an A subunit, B subunit and C subunit. Our focus of study is on the B subunit due to differences in function between the B56 family of proteins. There are five genes within the B56 family: B56α, B56β, B56ε, B56, and B56and while they are all within the same family, B56α, B56β, B56, and B56inhibit Wnt signaling while B56ε activates it. The goal of our research project is to identify where B56α and B56ε differ within their sequences to explain their differences in function in the Wnt pathway.

In working on our research project I was able to learn the process of research from start to finish. With the direction of Dr. Seeling, I spent time planning how and where the sequences of B56α and B56ε were to be “cut” with restriction enzymes using a computer program called Ape. In moving forward with our research, I learned various techniques that were needed such as how to set up restriction digests, gels for electrophoresis, gel purification, transformation, DNA purification, and other subcloning techniques. In addition, I practiced techniques such as microinjection into frog embryos which allowed me to work with our model organism, Xenopus laevis and learned much about their developmental process. Carrying out these techniques, which were part of a series of steps in gathering results, we were able to successfully subclone a DNA containing the N-terminus of the B56ε gene and the C-terminus and core of the B56α gene. Currently, the successfully cloned plasmid is ready to be used to synthesize RNA for microinjection and phenotypic analysis.

Working on identifying B56α and B56ε specificity I gained, not only the knowledge of how to carry out certain procedures and techniques, but the importance of being thorough, patient, and detail oriented. Being able to work alongside Dr. Seeling this summer on a research project taught me to acquire qualities such as these which has carried onto my classes and labs and will help me when I go onto more difficult subjects. As an aspiring orthodontist, researching allows for the opportunity to learn how to successfully apply the techniques I become proficient at and use them when looking for answers to a question within science.