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Transient Dominant Selection for the Modification and Generation of Recombinant Infectious Bronchitis Coronaviruses
We have developed a reverse genetics system for the avian coronavirus infectious bronchitis virus (IBV) in which a full-length cDNA corresponding to the IBV genome is inserted into the vaccinia virus genome under the control of a T7 promoter sequence. Vaccinia virus as a vector for the full-length I...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7121987/ https://www.ncbi.nlm.nih.gov/pubmed/25720477 http://dx.doi.org/10.1007/978-1-4939-2438-7_12 |
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author | Keep, Sarah M. Bickerton, Erica Britton, Paul |
author_facet | Keep, Sarah M. Bickerton, Erica Britton, Paul |
author_sort | Keep, Sarah M. |
collection | PubMed |
description | We have developed a reverse genetics system for the avian coronavirus infectious bronchitis virus (IBV) in which a full-length cDNA corresponding to the IBV genome is inserted into the vaccinia virus genome under the control of a T7 promoter sequence. Vaccinia virus as a vector for the full-length IBV cDNA has the advantage that modifications can be introduced into the IBV cDNA using homologous recombination, a method frequently used to insert and delete sequences from the vaccinia virus genome. Here, we describe the use of transient dominant selection as a method for introducing modifications into the IBV cDNA; this has been successfully used for the substitution of specific nucleotides, deletion of genomic regions, and the exchange of complete genes. Infectious recombinant IBVs are generated in situ following the transfection of vaccinia virus DNA, containing the modified IBV cDNA, into cells infected with a recombinant fowlpox virus expressing T7 DNA-dependent RNA polymerase. |
format | Online Article Text |
id | pubmed-7121987 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
record_format | MEDLINE/PubMed |
spelling | pubmed-71219872020-04-06 Transient Dominant Selection for the Modification and Generation of Recombinant Infectious Bronchitis Coronaviruses Keep, Sarah M. Bickerton, Erica Britton, Paul Coronaviruses Article We have developed a reverse genetics system for the avian coronavirus infectious bronchitis virus (IBV) in which a full-length cDNA corresponding to the IBV genome is inserted into the vaccinia virus genome under the control of a T7 promoter sequence. Vaccinia virus as a vector for the full-length IBV cDNA has the advantage that modifications can be introduced into the IBV cDNA using homologous recombination, a method frequently used to insert and delete sequences from the vaccinia virus genome. Here, we describe the use of transient dominant selection as a method for introducing modifications into the IBV cDNA; this has been successfully used for the substitution of specific nucleotides, deletion of genomic regions, and the exchange of complete genes. Infectious recombinant IBVs are generated in situ following the transfection of vaccinia virus DNA, containing the modified IBV cDNA, into cells infected with a recombinant fowlpox virus expressing T7 DNA-dependent RNA polymerase. 2014-12-18 /pmc/articles/PMC7121987/ /pubmed/25720477 http://dx.doi.org/10.1007/978-1-4939-2438-7_12 Text en © Springer Science+Business Media New York 2015 This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic. |
spellingShingle | Article Keep, Sarah M. Bickerton, Erica Britton, Paul Transient Dominant Selection for the Modification and Generation of Recombinant Infectious Bronchitis Coronaviruses |
title | Transient Dominant Selection for the Modification and Generation of Recombinant Infectious Bronchitis Coronaviruses |
title_full | Transient Dominant Selection for the Modification and Generation of Recombinant Infectious Bronchitis Coronaviruses |
title_fullStr | Transient Dominant Selection for the Modification and Generation of Recombinant Infectious Bronchitis Coronaviruses |
title_full_unstemmed | Transient Dominant Selection for the Modification and Generation of Recombinant Infectious Bronchitis Coronaviruses |
title_short | Transient Dominant Selection for the Modification and Generation of Recombinant Infectious Bronchitis Coronaviruses |
title_sort | transient dominant selection for the modification and generation of recombinant infectious bronchitis coronaviruses |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7121987/ https://www.ncbi.nlm.nih.gov/pubmed/25720477 http://dx.doi.org/10.1007/978-1-4939-2438-7_12 |
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