dc.contributor.author | Lopez-Galindez, Luis Cecilio | |
dc.contributor.author | Lopez, Juan Antonio | |
dc.contributor.author | Melero, Jose Antonio | |
dc.contributor.author | Fuente, Luis de la | |
dc.contributor.author | Martínez, Concepción | |
dc.contributor.author | Ortin, Juan | |
dc.contributor.author | Perucho, Manuel | |
dc.date.accessioned | 2019-06-05T10:51:20Z | |
dc.date.available | 2019-06-05T10:51:20Z | |
dc.date.issued | 1988-05 | |
dc.identifier.citation | Proc Natl Acad Sci U S A. 1988 May;85(10):3522-6. | es_ES |
dc.identifier.issn | 0027-8424 | es_ES |
dc.identifier.uri | http://hdl.handle.net/20.500.12105/7734 | |
dc.description.abstract | We have applied the RNase A mismatch cleavage method to analyze genetic variability in RNA viruses by using influenza virus as a model system. Uniformly labeled RNA probes synthesized from a cloned hemagglutinin gene of a given viral strain were hybridized to RNA isolated from other strains of characterized or uncharacterized genetic composition. The RNA.RNA heteroduplexes containing a variable number of base mismatches were digested with RNase A, and the resistant products were analyzed by denaturing polyacrylamide gel electrophoresis. We show that many of these single base mismatches are cleaved by RNase A, generating unique and characteristic patterns of resistant RNA fragments specific for each of the different viral strains. Comparative analysis of the cleavage patterns allows a qualitative estimation of the genetic relatedness and evolution of field strains. We also show that cleavage by RNase A at single base mismatches can readily detect and localize point mutations present in monoclonal antibody-resistant variants. This method should have wide applications in the study of RNA viruses, not only for epidemiological analysis but also in some diagnostic problems, such as characterization of phenotypic mutants. | es_ES |
dc.description.sponsorship | This work was supported by National Institutes of Health Grant CA33021 awarded by the Nationa l Cancer Institute to M.P. and by grants from the Comision Asesora de Investigacion Cientificay Tecnica (Grant 608/438) and Fondo de Investigaciones Sanitarias to J.O. and J.A.M.C.L.-G. was a recipient of a NATO short-term post doctoral fellow-ship while on leave from the Centro Nacional de Microbiologia, Majadahonda, Madrid | es_ES |
dc.language.iso | eng | es_ES |
dc.publisher | National Academy of Sciences | es_ES |
dc.type.hasVersion | VoR | es_ES |
dc.relation.isreferencedby | Proc Natl Acad Sci U S A. 1988 May;85(10):3522-6 | es_ES |
dc.rights.uri | http://creativecommons.org/licenses/by-nc-sa/4.0/ | * |
dc.subject.mesh | Animals | es_ES |
dc.subject.mesh | Cell Line | es_ES |
dc.subject.mesh | Chick Embryo | es_ES |
dc.subject.mesh | Influenza A virus | es_ES |
dc.subject.mesh | Nucleic Acid Hybridization | es_ES |
dc.subject.mesh | Plasmids | es_ES |
dc.subject.mesh | Ribonuclease, Pancreatic | es_ES |
dc.subject.mesh | Genes, Viral | es_ES |
dc.subject.mesh | Genetic Variation | es_ES |
dc.subject.mesh | Mutation | es_ES |
dc.title | Analysis of genetic variability and mapping of point mutations in influenza virus by the RNase A mismatch cleavage method | es_ES |
dc.type | journal article | es_ES |
dc.rights.license | Atribución-NoComercial-CompartirIgual 4.0 Internacional | * |
dc.identifier.pubmedID | 3368463 | es_ES |
dc.format.volume | 85 | es_ES |
dc.format.number | 10 | es_ES |
dc.format.page | 3522-6 | es_ES |
dc.contributor.funder | Instituto de Salud Carlos III | |
dc.identifier.journal | Proceedings of the National Academy of Sciences of the United States of America | es_ES |
dc.repisalud.centro | ISCIII::Centro Nacional de Microbiología | es_ES |
dc.repisalud.institucion | ISCIII | es_ES |
dc.rights.accessRights | open access | es_ES |