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Title: Selection of aptamers for signal transduction proteins and development of highly sensitive aptamer probes
Authors: TOK JUNIE
Keywords: aptamer, probe, selex, signal transduction protein, capillary electrophoresis, non-selex
Issue Date: 4-Dec-2009
Citation: TOK JUNIE (2009-12-04). Selection of aptamers for signal transduction proteins and development of highly sensitive aptamer probes. ScholarBank@NUS Repository.
Abstract: The development of aptamers from the improvement in selection technologies to its potential applications in diagnostics was explored in this thesis. High affinity aptamers for important signal transduction proteins, i.e. Cdc42-GTP, PAK1 and MRCKa were successfully selected in the low micro- to nanomolar range using non-SELEX, which is a new technique with capillary electrophoresis as an instrumental platform. The potential application of aptamers in cellular imaging was then studied. A representative MRCKa aptamer was used for the development of molecular aptamer beacon (MAB) through modification into a hairpin structure with a fluorescence-quenching pair at the 5? and 3? ends. In the presence of MRCKa, the MRCKa-binding conformer is preferred, giving rise to an increase in fluorescence. This simple method for MAB design could be potentially applied to other aptamers for modification into a protein probe. Preliminary studies showed that the aptamer retained its specificity for the protein in cellular environment.
Appears in Collections:Ph.D Theses (Open)

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