AN RNA APTAMER CONTAINING TWO BINDING SITES AGAINST THE HCV MINUS-IRES DOMAIN I |
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Authors: | Keisuke Konno Mana Iizuka Syusuke Fujita Satoshi Nishikawa Tsunemi Hasegawa Kotaro Fukuda |
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Affiliation: | 1. Department of Material and Biological Chemistry , Faculty of Science, Yamagata University , Yamagata, Japan;2. Age Dimension Research Center , National Institute of Advanced Industrial Science and Technology (AIST) , Tsukuba, Japan |
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Abstract: | The higher order structure of HCV (?)IRES containing five stem-loop structures (domain I) is essential for HCV replication because the viral RNA-dependent RNA polymerase, NS5B, recognizes it as the initiation site for plus-strand synthesis. To inhibit a de novo synthesis of plus-strand RNA molecules, in vitro selection against (?)IRES domain I was performed. One of the obtained aptamers, AP30, contained two consensus sequences within a random sequence region. Two consensus sequences form two apical loops and mutational analysis showed that both sequences were essential for binding to the target and for inhibiting NS5B-mediated RNA synthesis in vitro. |
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Keywords: | HCV minus-IRES NS5B in vitro selection RNA aptamer |
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