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Show 78 Sierra Wilen college of science Introduction: NMDARs, which are a type of glutamate receptor, have a significant role in neurodegenera-tive diseases such as Alzheimers and Parkinsons. In neurodegenerative diseases NMDARs are overstimu-lated. Overstimulation of these neuroreceptors causes the cell to die. Conus toxins found in snail venom are natural inhibitors of NMDARs. These toxins bind to a portion of NMDARs called S1-S2 subdomains. Analysis of conus toxin-S1S2 binding reactions may lead us to finding similar drugs that can cure neuro-logical diseases. Method & Results: The S1 and S2 encoding DNA portions were initially separated by unneeded DNA. My task during the 2011 Summer was to construct a linked S1-S2 encoding gene, as seen in the figure below [left]. This was done by a series of PCR reactions. S1 and S2 were amplified by doing two separate PCR reac-tions [lanes 1&2]. The clones were linked together through a third PCR reaction [lanes 3&4]. The gene was then inserted into a modified vector made by another student researcher. During the Fall 2011 semester, I expressed the protein using the modified vector. Temperature, IPTG concentration, inducing time, and many other conditions were tested. The result of doing multiple inductions that tested the various condi-tions was expressed insoluble protein, as seen the right-hand figure below. Future Plans: The next step in the experiment is to attain the protein in soluble form. The soluble form of the protein will be used in x-ray crystallography. X-ray crystallography will provide us with a molecular picture of the conus toxin-S1S2 interaction. SDS-page gel stained with Coomassie Blue. Agarose gel stained with ethidium bromide. STRUCTURE AND ACTIVITY OF GLUTAMATE RECEPTOR-TARGETED CONUS TOXINS Sierra Wilen (Martin Horvath) Department of Biology University of Utah UNDERGRADUATE RESEARCH ABSTRACTS Martin Horvath |