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dc.contributor.advisor Huang, Dr. Patrick
dc.contributor.advisor Tinnacher, Dr. Ruth
dc.creator Sin, Davina K.
dc.date 2018-05-31
dc.date.accessioned 2018-06-14T00:14:07Z
dc.date.available 2018-06-14T00:14:07Z
dc.date.issued 2018-06-13
dc.identifier.uri http://hdl.handle.net/10211.3/203802
dc.description.abstract Single stranded oligonucleotides called aptamers that have a binding affinity to reduced glutathione (GSH) and oxidized glutathione (GSSG) were isolated. An iterative technique called Systematic Evolution of Ligands by Exponential Enrichment (SELEX) was used to obtain an enriched aptamer pool of randomized sequences for GSH and another enriched aptamer pool for GSSG. Quantification via UV-visible spectroscopy yielded an aptamer pool with a nucleic acid concentration of 610.4 ng/L for GSH and 660.4 ng/L for GSSG. The resulting amplicons were inserted into E. coli for bacterial cloning to allow for DNA extraction and sequencing. Twenty-four GSH-binding aptamers and twenty-four GSSG-binding aptamers were isolated and sequenced. The GSH-binding aptamers are sensitive to as low as a 0.1 mM GSH solution while the GSSG-binding aptamers are sensitive to as low as a 0.01 mM GSSG solution. The Unified Nucleic Acid Folding Mfold web server was used to predict aptamer structures. The aptamers were analyzed for consistent sequences and structural motifs. Future studies should focus on testing the obtained aptamers with fluorescence binding assay to determine the sequence and structural motif with the highest binding affinity to GSH and GSSG respectively. en_US
dc.language English en_US
dc.subject Aptamers, Nucleotide en_US
dc.subject Glutathione en_US
dc.title Isolation of Aptamers with Binding Affinity for Reduced and Oxidized Glutathione en_US
dc.type Thesis en_US
dc.contributor.primaryAdvisor Halim, Dr. Marlin
thesis.degree.name Master of Science in Chemistry with Biochemistry Option en_US

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