KMID : 0380020050200020088
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Korean Journal of Biotechnology and Bioengineering 2005 Volume.20 No. 2 p.88 ~ p.92
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Development and Characterization of RNA Aptamers for Phosphorylated Amino Acids
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Cho Soo-Hyung
Kim Byung-Gee
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Abstract
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Phosphorylation of amino acid residues in proteins, plays a major role in biological mechanism. Phosphorylation acts as a process regulating the protein activity in variable pathways such as metabolism, signal transduction and cell division. Therefore the development of ligands for phosphoamino acids are an important work for protein analysis and proteomics studies. In this study, RNA aptamers for o-phosphoserine, o-phosphotyrosine and o-phosphotyrosine which appears frequently in nature were developed by in vitro evolution method. We could obtain similar sequences from random RNAs of 40 mer by SELEX method through 10 cycles. As result, the aptamers for o-phosphoserine and o-phosphothreonine among phosphoamino acids aptamers showed high affinity of Kd=2.60 nM and 2.65 nM for their target molecules, respectively. In addition, these aptamers could be confirmed the high selectivity for their target.
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KEYWORD
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Phosphorylation, phosphoamino acids, aptamer, SELEX
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