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http://hdl.handle.net/11375/22784
Title: | Creation and Characterization of a Fluorescence Signalling DNA Enzyme |
Other Titles: | A Fluorescence Signalling DNA Enzyme |
Authors: | Mei, Shirley |
Advisor: | Li, Yingfu |
Department: | Biochemistry |
Keywords: | fluorescence;DNA;enzyme;in vitro |
Publication Date: | Sep-2003 |
Abstract: | ππ― π·πͺπ΅π³π° selection has been widely used to isolate single-stranded DNA molecules from large random-sequence pools that are able to perform a desired catalytic reaction, or bind to a target molecule. Using this technique, we have created a DNA enzyme, named DET22-18, which has a uniquely linked chemical catalysis/real-time signaling capability. It is a true enzyme with a π¬cββ of~7 minβ»ΒΉ -the second fastest rate ever reported for a DNA enzyme. The DNA enzyme cleaves a substrate containing a single ribonucleotide linkage embedded in a DNA chain and sandwiched between a fluorophore-labeled deoxyribonucleotide and a quencher-modified deoxyribonucleotide. The ability of DET22-18 to generate a large fluorescence signal provides a useful tool to engineer potential allosteric deoxyribozyme biosensors for real-time detection of important biological targets. To provide a proof of concept that a reporter system can be built from the above signaling DNA enzyme, the cis-acting version of this enzyme, DEC22-18A, was engineered into an allosterically regulated deoxyribozyme biosensor that can report ATP. A preliminary investigation was also conducted to determine a possible secondary structure of the DNA enzyme. This study lays a foundation for pursuing novel signaling DNA enzymes for biological detection directed applications. |
URI: | http://hdl.handle.net/11375/22784 |
Appears in Collections: | Digitized Open Access Dissertations and Theses |
Files in This Item:
File | Description | Size | Format | |
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mei_shirley_2003Sept_masters.pdf | 16.4 MB | Adobe PDF | View/Open |
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