Presentation description

In the Brasch Lab, we study the molecular mechanism behind formation of neuronal synapses, with specific focus on the structure and function of the protein complexes assembling in the extracellular space. Previous research suggests that proper protein interactions are crucial for the overall structure and stability of the synapses, and any disturbances could result in neurological diseases.

My goal in the Brasch lab is to design a recombinant mouse neurexin DNA to express neurexin in mammalian cells for downstream cell aggregation assays to study protein interactions. This recombinant expression DNA will help further experiments that seek to study the synaptic protein interactome of Neurexins. In patients with schizophrenia, neurexin, a synaptic adhesion protein, is found to be mutated. I mutated vector DNA using PCR to introduce a restriction enzyme site that allows us to cut the vector and insert neurexin DNA. We used PCR to produce DNA encoding neurexin flanked by the same restriction enzyme sites such that the insert can fit into the prepared vector and 'glued' these together through a ligation enzyme.

We have been able to successfully mutate the vector DNA using PCR and insert our recombinant DNA. Our future directions, utilizes ligation reaction to attach the insert into cut vector DNA.

This study will provide recombinant expression neurexin DNA needed for cell expression to properly study this important synaptic protein.