Iterative Site-Directed Mutagenesis Towards Directed Evolution of Cathepsin-L

In our research, the process of iterative site-directed mutagenesis will be used in order to direct evolution of Cathepsin-L. This protein is an important lysosomal endopeptidase enzyme, which is important for the initiation of the process of protein degradation. The goal of our research is to select specific sites of the protein to mutate in order to observe the phenotypic outcomes. Once different mutants of Cathepsin-L are created, we will be able to study the difference in the functionality of the protein, such as whether the mutation hindered or enabled the protein’s ability to perform.

I expect to not only sharpen my pre-existing laboratory and research skills, but also gain new ones. It has been such a valuable experience to be able to perform actual techniques that I have only learned about in school. By being able to do things hands on, I have gained a better understanding of many concepts. Throughout this program, I have made antibiotic containing plates, performed chemical transformations with E. coli, performed plasmid mini-preps towards DNA analysis, as well as protein purification and isolation through affinity chromatography. All of these are techniques and processes that are important for  me to understand and be able to do, not only as a student, but as a scientist.

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