SELECTION OF HIGH-AFFINITY RNA LIGANDS TO REVERSE-TRANSCRIPTASE - INHIBITION OF CDNA SYNTHESIS AND RNASE-H ACTIVITY
SELECTION OF HIGH-AFFINITY RNA LIGANDS TO REVERSE-TRANSCRIPTASE - INHIBITION OF CDNA SYNTHESIS AND RNASE-H ACTIVITY
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DOI：
10.1021/bi00195a016
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发表时间：
1994-07-26
期刊：
影响因子：
2.9
通讯作者：
GOLD, L
中科院分区：
文献类型：
--
作者：
CHEN, H;GOLD, L
利用SELEX（系统进化的配体通过指数富集）程序从组合RNA文库中分离出特异性的、高亲和力的RNA配体，用于禽成髓细胞病病毒和Moloney小鼠白血病病毒逆转录酶。所选择的RNA配体以大约纳摩尔的解离常数结合各自的逆转录酶。这些配体在不同靶酶的选择中没有表现出一级序列的保守性。此外，所选择的配体与模板/引物复合物的结合竞争，抑制rna依赖的DNA聚合酶和同源逆转录酶的RNase H活性。SELEX可以产生高亲和力和高特异性的寡核苷酸拮抗剂来对抗蛋白质家族的特定成员。
Specific, high-affinity RNA ligands to avian myeloblastosis virus and Moloney murine leukemia virus reverse transcriptases were isolated from a combinatorial RNA library using the SELEX (systematic evolution of ligands by exponential enrichment) procedure. The selected RNA ligands bound their respective reverse transcriptases with approximately nanomolar dissociation constants. The ligands did not exhibit primary sequence conservation from selections against different target enzymes. Moreover, the selected ligands competed with the binding of template/primer complex and inhibited both the RNA-dependent DNA polymerase and the RNase H activities of the cognate reverse transcriptase. SELEX can yield both high-affinity and high-specificity oligonucleotide antagonists against specific members of a protein family.