使用分段同位素标记对100 kDa丙型肝炎病毒内部核糖体进入位点RNA进行核磁共振研究。

NMR study of 100 kDa HCV IRES RNA using segmental isotope labeling.

作者信息

Kim Insil, Lukavsky Peter J, Puglisi Joseph D

机构信息

Department of Structural Biology, School of Medicine, Stanford University, California 94305-5126, USA.

出版信息

J Am Chem Soc. 2002 Aug 14;124(32):9338-9. doi: 10.1021/ja026647w.

DOI:10.1021/ja026647w

PMID:12167005

Abstract

RNA NMR is hindered by the large size of most biological RNAs. We present here a simple method for segmental isotopic labeling of an RNA fragment within the context of a larger RNA. The methodology uses transcription and ribozyme cleavage to prepare appropriate ends for RNA ligase catalyzed ligation. We demonstrate that a 64 nucleotide domain of the Hepatitis C virus internal ribosome entry site (IRES) RNA adopts an independently folded domain within the context of the intact, 100 kDa IRES.

摘要

RNA核磁共振技术受到大多数生物RNA分子量大的限制。我们在此展示了一种在较大RNA背景下对RNA片段进行分段同位素标记的简单方法。该方法利用转录和核酶切割来制备适合RNA连接酶催化连接的末端。我们证明，丙型肝炎病毒内部核糖体进入位点（IRES）RNA的一个64个核苷酸的结构域在完整的100 kDa IRES背景下形成了一个独立折叠的结构域。