CHED 858 |
| Our previously reported 2'-5' RNA-branching deoxyribozymes, such as 7S11 and 10DM24, have allowed for the development of new techniques for studying large RNA molecules. To expand this focus, we seek to identify unique deoxyribozymes that individually have the ability to catalyze the formation of 2',5'-branched linkages between a single set of the four different combinations of nucleic acid substrates (RNA-RNA, RNA-DNA, DNA-RNA, and DNA-DNA). As a set, these deoxyribozymes will allow for a generalized method of synthesizing branched nucleic acid structures. We designed various in vitro selection experiments to identify individual deoxyribozymes capable of catalyzing these reactions. Each selection focused on identifying a deoxyribozyme capable of catalyzing one of the four different ligations. During these studies, reaction conditions were altered for some selections to select for faster enzymes that could also tolerate more biologically relevant conditions. When a selection pool was dominated by sequences capable of catalyzing the desired reaction, the pool was cloned and screened for enzymes with not only fast activity, but also high yield. The resulting deoxyribozymes will be characterized thoroughly to determine possible sequence requirements, to give insight into the catalytic function of these branch-forming deoxyribozymes, and to contribute to the collection of deoxyribozymes capable of synthesizing branched nucleic acids. |
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Undergraduate Research Poster Session: Biochemistry
2:00 PM-4:00 PM, Monday, April 7, 2008 Morial Convention Center -- Hall A, Poster
Division of Chemical Education |