Date of this Version
Zalman, C. & Vecchio, A. (2020) Mutagenesis of Claudins to Probe in vivo Interactions and Assemblies. Poster prtesentation, UNL Spring Research Fair, University of Nebraska-Lincoln.
Paracellular transport of solutes and the control of the flow of molecules through the intracellular space in vertebrate epithelia is directed by tight junctions (TJs). Claudins form paracellular barriers and pores that determine tight junction permeability. This investigation attempts to explain the molecular bases for destruction and reconstruction of tight junctions within epithelial cells, occurring via both natural and disease-causing mechanisms. This interdisciplinary research is important in the advancement of our understanding of human biology and health, as different disruptions to epithelial tight junctions are hallmarks of many human diseases. The overall objective was to take previously made Claudin containing vectors (pEGFP-N3) containing a Green Fluorescent Protein (GFP) sequence and utilize site specific mutations to change the sequences to express Cyan Fluorescent Protein (CFP) and Yellow Fluorescent Protein (YFP). Proteins tested in this experiment were Human Claudins 1, 9, 19, Tricellulin, and Occludin. Once the specified claudins and TAMPs are tagged with CFP and YFP, FRET microscopy techniques can be utilized to determine generic interactions within the formation of TJs, forming a baseline for future research.