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...
| Authors: | ; ; ; ; ; ; |
|---|---|
| Format: | Article |
| Publication date: | 1988 |
| Country: | España |
| Institution: | Instituto de Salud Carlos III (ISCIII) |
| Repository: | Repisalud |
| Language: | English |
| OAI Identifier: | oai:repisalud.isciii.es:20.500.12105/7734 |
| Online Access: | http://hdl.handle.net/20.500.12105/7734 |
| Access Level: | Open access |
| Keyword: | Animals Cell Line Chick Embryo Influenza A virus Nucleic Acid Hybridization Plasmids Ribonuclease, Pancreatic Genes, Viral Genetic Variation Mutation |
| _version_ | 1878737005306707968 |
|---|---|
| author | Lopez-Galindez, Luis Cecilio |
| 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 |
| author_browse | Fuente, Luis de la Lopez, Juan Antonio Lopez-Galindez, Luis Cecilio Martínez, Concepción Melero, Jose Antonio Ortin, Juan Perucho, Manuel |
| 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 |
| collection | Repisalud |
| 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. |
| eu_rights_str_mv | openAccess |
| format | article |
| id | ES_98ec2c19d2ce00196ead1bd3fdb55835 |
| institution | Instituto de Salud Carlos III (ISCIII) |
| instname_str | Instituto de Salud Carlos III (ISCIII) |
| language | eng |
| network_acronym_str | ES |
| network_name_str | España |
| oai_identifier_str | oai:repisalud.isciii.es:20.500.12105/7734 |
| publishDate | 1988 |
| publishDateSort | 1988 |
| publisher | National Academy of Sciences |
| publisherStr | National Academy of Sciences |
| reponame_str | Repisalud |
| spelling | Analysis of genetic variability and mapping of point mutations in influenza virus by the RNase A mismatch cleavage method Lopez-Galindez, Luis Cecilio Lopez, Juan Antonio Melero, Jose Antonio Fuente, Luis de la Martínez, Concepción Ortin, Juan Perucho, Manuel Animals Cell Line Chick Embryo Influenza A virus Nucleic Acid Hybridization Plasmids Ribonuclease, Pancreatic Genes, Viral Genetic Variation Mutation 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. National Academy of Sciences http://hdl.handle.net/20.500.12105/7734 |
| 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 | 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_short | 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 |
| topic | Animals Cell Line Chick Embryo Influenza A virus Nucleic Acid Hybridization Plasmids Ribonuclease, Pancreatic Genes, Viral Genetic Variation Mutation |
| topic_facet | Animals Cell Line Chick Embryo Influenza A virus Nucleic Acid Hybridization Plasmids Ribonuclease, Pancreatic Genes, Viral Genetic Variation Mutation |
| url | http://hdl.handle.net/20.500.12105/7734 |