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Title:Identification of Deoxyribozymes with Nucleopeptide Linkage Formation and Phenylalanine Ammonia Lyase Activities
Author(s):Li, Xinyi
Contributor(s):Silverman, Scott K.
nucleopeptide linkage
phenylalanine ammonia lyase
Abstract:Biocatalysts such as deoxyribozymes (single-strand DNA with catalytic activity) could be very useful tools in the many areas of study. Here we present two projects for in vitro identification of deoxyribozymes for specific chemical reactions. One is for nucleopeptide linkage formation, the other is for phenylalanine lyase activity. Nucleopeptide conjugation is involved in many activities in nature including topoisomerization. In our study, we focus on topoisomerase-related polypeptide sequence and aimed to identify deoxyribozymes with the ability to catalyze tyrosine-oligonucleotides linkage. If found, the deoxyribozyme could be useful for the study of Tyrosyl-DNA phosphodiesterase, an enzyme for breaking the trapped topoisomerase-oligonucleotides bond. Phenylalanine lyase is an enzyme exists in plants that turns phenylalanine into trans-cinamic acid. It is studied as a drug candidate for Phenylketonuria. The deoxyribozymes for this activity could help the study. Both target deoxyribozymes would be identified from randomized pool with single-strand DNA in various length (30nt ~ 80nt). The pool was first ligated with 5’ imidazole DNA-RNA hybridized substrate or 3’ phenylalanine DNA substrate depended on the target reaction. The ligated pool then go through selection under the condition of targeted reaction. The product was captured using either copper(I)-catalyzed azide/alkyne cycloaddition “click” reaction or DTT treatment following iodoacetate-oligonucleotides capture reaction depended on which deoxyribozyme was selected. Both experiments are still ongoing.
Issue Date:2017
Genre:Dissertation / Thesis
Rights Information:Copyright 2017 Xinyi Li
Date Available in IDEALS:2017-05-04

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