NMR Study of 100 KDa HCV IRES RNA Using Segmental Isotope Labeling

Overview

Journal J Am Chem Soc

Publisher American Chemical Society

Specialty Chemistry

Date 2002 Aug 9

PMID 12167005

Citations 45

Authors

Insil Kim

Peter J Lukavsky

Joseph D Puglisi

Affiliations

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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.

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