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In vitro Selection of Ligase Ribozymes Containing 2'-Amino Groups

机译:含2'-氨基的连接酶核酶的体外选择

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摘要

Novel ribozymes containing 2'-amino groups in the side chains were in vitro selected to accelerate their ligation reaction rates with oligodeoxynucleotides. The ligation rate of random sequenced RNAs in the starting pool was accelerated by incorporation of 2'-amino-2'-deoxyuridine and N~6-(6-aminohexyl) adenosine. The incorporation of the amino group enhanced the activity of non-selected RNAs independent of the incorporation site. In vitro selection using 2'-amino-2'-deoxyuridine instead of uridine produced more active ribozymes. In this case, the activity of ribozymer was reduced when N~6-(6-aminohexyl)adenosine was incorporated into the selected RNAs instead of natural adenosine. The presence of amino groups as well as the incorporation site affected the activity of the in vitro selected ribozyme. It seems that RNAs with tertiary structures suitable for the ligation reaction were selected by the in vitro method.
机译:在体外选择了在侧链中含有2'-氨基的新型核酶,以加速它们与寡脱氧核苷酸的连接反应速率。通过掺入2'-氨基-2'-脱氧尿苷和N〜6-(6-氨基己基)腺苷可加快起始库中随机测序RNA的连接速度。氨基的掺入增强了独立于掺入位点的未选择RNA的活性。使用2'-氨基-2'-脱氧尿苷代替尿苷进行体外选择产生了更多的活性核酶。在这种情况下,当将N〜6-(6-氨基己基)腺苷而非天然腺苷掺入所选的RNA中时,核酶的活性降低。氨基的存在以及掺入位点影响了体外选择的核酶的活性。似乎通过体外方法选择了适合于连接反应的三级结构的RNA。

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