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http://dspace.mediu.edu.my:8181/xmlui/handle/10261/4366| Title: | Structural analysis of hepatitis C RNA genome using DNA microarrays |
| Authors: | European Commission Ministerio de Ciencia y Tecnología (España) Instituto Nacional de Técnica Aeroespacial (España) Comunidad de Madrid |
| Publisher: | Oxford University Press |
| Description: | PMCID: PMC443556.-- Final edited version available at: http://dx.doi.org/10.1093/nar/gnh088 Many studies have tried to identify specific nucleotide sequences in the quasispecies of hepatitis C virus (HCV) that determine resistance or sensitivity to interferon (IFN) therapy, unfortunately without conclusive results. Although viral proteins represent the most evident phenotype of the virus, genomic RNA sequences determine secondary and tertiary structures which are also part of the viral phenotype and can be involved in important biological roles. In this work, a method of RNA structure analysis has been developed based on the hybridization of labelled HCV transcripts to microarrays of complementary DNA oligonucleotides. Hybridizations were carried out at non-denaturing conditions, using appropriate temperature and buffer composition to allow binding to the immobilized probes of the RNA transcript without disturbing its secondary/tertiary structural motifs. Oligonucleotides printed onto the microarray covered the entire 5' non-coding region (5'NCR), the first three-quarters of the core region, the E2–NS2 junction and the first 400 nt of the NS3 region. We document the use of this methodology to analyse the structural degree of a large region of HCV genomic RNA in two genotypes associated with different responses to IFN treatment. The results reported here show different structural degree along the genome regions analysed, and differential hybridization patterns for distinct genotypes in NS2 and NS3 HCV regions. M.M. was supported in part by grants DITTO-HCV QLK2-2000-0836 from the European Commission FP5 and Red Nacional de Gastroenterología y Hepatología (C03/02). The work in Hospital Vall d’Hebron was supported in part by grants SAF2000/0183 and PROFIT (FIT-010000-2003-51) from the Ministerio de Ciencia y Tecnología. The work at CAB was supported by the European Union, Instituto Nacional de Técnica Aerospacial, Ministerio de Ciencia y Tecnología and Comunidad de Madrid. Peer reviewed |
| URI: | http://dspace.mediu.edu.my:8181/xmlui/handle/10261/4366 |
| Other Identifiers: | Nucleic Acids Res. 2004; 32(11): e90. http://dx.doi.org/10.1093/nar/gnh088 0305-1048 (Print) 1362-4962 (Online) http://hdl.handle.net/10261/4366 10.1093/nar/gnh088 |
| Appears in Collections: | Digital Csic |
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