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dc.contributor.authorLopez-Galindez, Luis Cecilio 
dc.contributor.authorLopez, Juan Antonio 
dc.contributor.authorMelero, Jose Antonio 
dc.contributor.authorFuente, Luis de la 
dc.contributor.authorMartinez, C 
dc.contributor.authorOrtin, J
dc.contributor.authorPerucho, M
dc.identifier.citationProc Natl Acad Sci U S A. 1988 May;85(10):3522-6.es_ES
dc.description.abstractWe 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.sponsorshipThis 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, Madrides_ES
dc.publisherNational Academy of Scienceses_ES
dc.relation.isversionofPublisher's versiones_ES
dc.relation.isreferencedbyProc Natl Acad Sci U S A. 1988 May;85(10):3522-6es_ES
dc.subject.meshAnimals es_ES
dc.subject.meshCell Line es_ES
dc.subject.meshChick Embryo es_ES
dc.subject.meshInfluenza A virus es_ES
dc.subject.meshNucleic Acid Hybridization es_ES
dc.subject.meshPlasmids es_ES
dc.subject.meshRibonuclease, Pancreatic es_ES
dc.subject.meshGenes, Viral es_ES
dc.subject.meshGenetic Variation es_ES
dc.subject.meshMutation es_ES
dc.titleAnalysis of genetic variability and mapping of point mutations in influenza virus by the RNase A mismatch cleavage methodes_ES
dc.rights.licenseAtribución-NoComercial-CompartirIgual 4.0 Internacional*
dc.contributor.funderFondo de Investigaciones Sanitariases_ES
dc.identifier.journalProceedings of the National Academy of Sciences of the United States of Americaes_ES
dc.repisalud.centroISCIII::Centro Nacional de Microbiologíaes_ES

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