NMR study of 100 kDa HCV IRES RNA using segmental isotope labeling.
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.
Kim
Insil
I
Department of Structural Biology, School of Medicine, Stanford University, California 94305-5126, USA.
Lukavsky
Peter J
PJ
Puglisi
Joseph D
JD
D016174:Hepacivirus / Q000235:genetics*; D007553:Isotope Labeling; D019906:Nuclear Magnetic Resonance, Biomolecular; D012367:RNA, Viral / Q000737:chemistry*
D012367:RNA, Viral