大型RNA的互补片段标记：用于测量更多剩余偶极耦合（RDC）的经济制备方法及简化的核磁共振（NMR）谱图

Complementary segmental labeling of large RNAs: economic preparation and simplified NMR spectra for measurement of more RDCs.

作者信息

Tzakos Andreas G, Easton Laura E, Lukavsky Peter J

机构信息

MRC, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, United Kingdom.

出版信息

J Am Chem Soc. 2006 Oct 18;128(41):13344-5. doi: 10.1021/ja064807o.

DOI:10.1021/ja064807o

PMID:17031932

Abstract

NMR structure determination of large RNAs is often restricted by limited RDC information caused by chemical shift degeneracy. We established a general, time- and cost-effective methodology for the preparation of 13C/15N complementary labeled RNAs from a single plasmid. Applying this method to the 25 kDa BC1-DTE RNA, we were able to resolve severe chemical shift degeneracy, thereby almost doubling the number of RDC restraints in comparison to the conventional 13C,15N uniform-labeled RNA.

摘要

大RNA的核磁共振结构测定常常受到化学位移简并导致的剩余偶极耦合（RDC）信息有限的限制。我们建立了一种通用的、具有时间和成本效益的方法，用于从单个质粒制备13C/15N互补标记的RNA。将该方法应用于25 kDa的BC1-DTE RNA，我们能够解决严重的化学位移简并问题，从而使RDC约束的数量相比传统的13C、15N均匀标记RNA几乎增加了一倍。