Analysis of genetic variability and mapping of point mutations in influenza virus by the RNase A mismatch cleavage method

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 chara...

Descripción completa

Detalles Bibliográficos
Autores: Lopez-Galindez, Luis Cecilio, Lopez, Juan Antonio, Melero, Jose Antonio, Fuente, Luis de la, Martínez, Concepción, Ortin, Juan, Perucho, Manuel
Tipo de recurso: artículo
Fecha de publicación:1988
País:España
Institución:Instituto de Salud Carlos III (ISCIII)
Repositorio:Repisalud
Idioma:inglés
OAI Identifier:oai:repisalud.isciii.es:20.500.12105/7734
Acceso en línea:http://hdl.handle.net/20.500.12105/7734
Access Level:acceso abierto
Palabra clave:Animals
Cell Line
Chick Embryo
Influenza A virus
Nucleic Acid Hybridization
Plasmids
Ribonuclease, Pancreatic
Genes, Viral
Genetic Variation
Mutation
id ES_98ec2c19d2ce00196ead1bd3fdb55835
oai_identifier_str oai:repisalud.isciii.es:20.500.12105/7734
network_acronym_str ES
network_name_str España
repository_id_str
spelling Analysis of genetic variability and mapping of point mutations in influenza virus by the RNase A mismatch cleavage methodLopez-Galindez, Luis CecilioLopez, Juan AntonioMelero, Jose AntonioFuente, Luis de laMartínez, ConcepciónOrtin, JuanPerucho, ManuelAnimalsCell LineChick EmbryoInfluenza A virusNucleic Acid HybridizationPlasmidsRibonuclease, PancreaticGenes, ViralGenetic VariationMutationWe 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.National Academy of SciencesInstituto de Salud Carlos III20192019-06-0519881988-05-0119881988-05-01journal articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/20.500.12105/7734reponame:Repisaludinstname:Instituto de Salud Carlos III (ISCIII)Inglésengopen accesshttp://purl.org/coar/access_right/c_abf2Atribución-NoComercial-CompartirIgual 4.0 Internacionalhttp://creativecommons.org/licenses/by-nc-sa/4.0/info:eu-repo/semantics/openAccessoai:repisalud.isciii.es:20.500.12105/77342026-06-12T12:43:37Z
dc.title.none.fl_str_mv Analysis of genetic variability and mapping of point mutations in influenza virus by the RNase A mismatch cleavage method
title Analysis of genetic variability and mapping of point mutations in influenza virus by the RNase A mismatch cleavage method
spellingShingle Analysis of genetic variability and mapping of point mutations in influenza virus by the RNase A mismatch cleavage method
Lopez-Galindez, Luis Cecilio
Animals
Cell Line
Chick Embryo
Influenza A virus
Nucleic Acid Hybridization
Plasmids
Ribonuclease, Pancreatic
Genes, Viral
Genetic Variation
Mutation
title_short Analysis of genetic variability and mapping of point mutations in influenza virus by the RNase A mismatch cleavage method
title_full Analysis of genetic variability and mapping of point mutations in influenza virus by the RNase A mismatch cleavage method
title_fullStr Analysis of genetic variability and mapping of point mutations in influenza virus by the RNase A mismatch cleavage method
title_full_unstemmed Analysis of genetic variability and mapping of point mutations in influenza virus by the RNase A mismatch cleavage method
title_sort Analysis of genetic variability and mapping of point mutations in influenza virus by the RNase A mismatch cleavage method
dc.creator.none.fl_str_mv Lopez-Galindez, Luis Cecilio
Lopez, Juan Antonio
Melero, Jose Antonio
Fuente, Luis de la
Martínez, Concepción
Ortin, Juan
Perucho, Manuel
author Lopez-Galindez, Luis Cecilio
author_facet Lopez-Galindez, Luis Cecilio
Lopez, Juan Antonio
Melero, Jose Antonio
Fuente, Luis de la
Martínez, Concepción
Ortin, Juan
Perucho, Manuel
author_role author
author2 Lopez, Juan Antonio
Melero, Jose Antonio
Fuente, Luis de la
Martínez, Concepción
Ortin, Juan
Perucho, Manuel
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Instituto de Salud Carlos III
dc.subject.none.fl_str_mv Animals
Cell Line
Chick Embryo
Influenza A virus
Nucleic Acid Hybridization
Plasmids
Ribonuclease, Pancreatic
Genes, Viral
Genetic Variation
Mutation
topic Animals
Cell Line
Chick Embryo
Influenza A virus
Nucleic Acid Hybridization
Plasmids
Ribonuclease, Pancreatic
Genes, Viral
Genetic Variation
Mutation
description 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.
publishDate 1988
dc.date.none.fl_str_mv 1988
1988-05-01
1988
1988-05-01
2019
2019-06-05
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/20.500.12105/7734
url http://hdl.handle.net/20.500.12105/7734
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Atribución-NoComercial-CompartirIgual 4.0 Internacional
http://creativecommons.org/licenses/by-nc-sa/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Atribución-NoComercial-CompartirIgual 4.0 Internacional
http://creativecommons.org/licenses/by-nc-sa/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv National Academy of Sciences
publisher.none.fl_str_mv National Academy of Sciences
dc.source.none.fl_str_mv reponame:Repisalud
instname:Instituto de Salud Carlos III (ISCIII)
instname_str Instituto de Salud Carlos III (ISCIII)
reponame_str Repisalud
collection Repisalud
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869414232242520064
score 15.81155

Similares en LA Referencia