Green L, Waugh S, Binkley J P, Hostomska Z, Hostomsky Z, Tuerk C
NeXagen, Inc., Boulder, CO 80301.
J Mol Biol. 1995 Mar 17;247(1):60-8. doi: 10.1006/jmbi.1994.0122.
We had previously used in vitro RNA selection techniques to describe a consensus RNA pseudoknot that binds and inhibits HIV-1 reverse transcriptase (HIV-RT). In this work we constructed variants of this consensus pseudoknot in order to evaluate the contributions of individual nucleotide identities and secondary structure to affinity for HIV-RT. We have also used chemical modification of ligand RNAs to corroborate the predicted structure of the pseudoknot, to discover which modifiable groups are protected from chemical attack when bound to HIV-RT, and to find which modifications interfere with binding to HIV-RT. A novel interference study is presented which involves selection of ligands from a pool created by mixed reagent oligonucleotide synthesis in order to rapidly determine allowed substitutions of 2'-OCH3 groups for the usual 2'-OH group in such RNA ligands.
我们之前使用体外RNA筛选技术描述了一种能结合并抑制HIV-1逆转录酶（HIV-RT）的共有RNA假结。在这项工作中，我们构建了这种共有假结的变体，以评估单个核苷酸序列和二级结构对与HIV-RT亲和力的贡献。我们还对配体RNA进行化学修饰，以证实假结的预测结构，发现哪些可修饰基团在与HIV-RT结合时免受化学攻击，并找出哪些修饰会干扰与HIV-RT的结合。本文提出了一项新颖的干扰研究，该研究涉及从混合试剂寡核苷酸合成产生的文库中筛选配体，以便快速确定此类RNA配体中通常的2'-OH基团被2'-OCH3基团取代的允许情况。
