Rescue of recombinant zika virus from a bacterial artificial chromosome cdna clone

© 2019 Journal of Visualized Experiments.

Detalles Bibliográficos
Autores: Ávila-Pérez, Ginés, Park, Jun-Gyu, Nogales, Aitor, Almazán, Fernando, Martínez-Sobrido, Luis
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2019
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/241180
Acceso en línea:http://hdl.handle.net/10261/241180
Access Level:acceso abierto
Palabra clave:Immunology and Infection
Issue 148
Arbovirus
Flavivirus
Zika virus
Reverse genetics
Full-length cDNA infectious clones
Bacterial artificial chromosomes
cDNA transfection
Virus rescue
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spelling Rescue of recombinant zika virus from a bacterial artificial chromosome cdna cloneÁvila-Pérez, GinésPark, Jun-GyuNogales, AitorAlmazán, FernandoMartínez-Sobrido, LuisImmunology and InfectionIssue 148ArbovirusFlavivirusZika virusReverse geneticsFull-length cDNA infectious clonesBacterial artificial chromosomescDNA transfectionVirus rescue© 2019 Journal of Visualized Experiments.The association of Zika virus (ZIKV) infection with neurological complications during the recent worldwide outbreak and the lack of approved vaccines and/or antivirals have underscored the urgent need to develop ZIKV reverse genetic systems to facilitate the study of ZIKV biology and the development of therapeutic and/or prophylactic approaches. However, like with other flaviviruses, the generation of ZIKV full-length infectious cDNA clones has been hampered due to the toxicity of viral sequences during its amplification in bacteria. To overcome this problem, we have developed a nontraditional approach based on the use of bacterial artificial chromosomes (BACs). Using this approach, the full-length cDNA copy of the ZIKV strain Rio Grande do Norte Natal (ZIKV-RGN) is generated from four synthetic DNA fragments and assembled into the single-copy pBeloBAC11 plasmid under the control of the human cytomegalovirus (CMV) immediate-early promoter. The assembled BAC cDNA clone is stable during propagation in bacteria, and infectious recombinant (r)ZIKV is recovered in Vero cells after transfection of the BAC cDNA clone. The protocol described here provides a powerful technique for the generation of infectious clones of flaviviruses, including ZIKV, and other positive-strand RNA viruses, particularly those with large genomes that have stability problems during bacterial propagation.This work was supported in part by grants from the Spanish Ministry of Economy and Competitiveness (MINECO, grant number BFU2016-79127-R) to F.A.T. and the National Institutes of Health (NIH, grant number 1R21AI120500) to L.M.S. and F.A.T.MYJoVE CorporationMinisterio de Economía y Competitividad (España)National Institutes of Health (US)Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2021202120192021info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/241180reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BFU2016-79127-Rhttp://dx.doi.org/10.3791/59537Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2411802026-05-22T06:33:51Z
dc.title.none.fl_str_mv Rescue of recombinant zika virus from a bacterial artificial chromosome cdna clone
title Rescue of recombinant zika virus from a bacterial artificial chromosome cdna clone
spellingShingle Rescue of recombinant zika virus from a bacterial artificial chromosome cdna clone
Ávila-Pérez, Ginés
Immunology and Infection
Issue 148
Arbovirus
Flavivirus
Zika virus
Reverse genetics
Full-length cDNA infectious clones
Bacterial artificial chromosomes
cDNA transfection
Virus rescue
title_short Rescue of recombinant zika virus from a bacterial artificial chromosome cdna clone
title_full Rescue of recombinant zika virus from a bacterial artificial chromosome cdna clone
title_fullStr Rescue of recombinant zika virus from a bacterial artificial chromosome cdna clone
title_full_unstemmed Rescue of recombinant zika virus from a bacterial artificial chromosome cdna clone
title_sort Rescue of recombinant zika virus from a bacterial artificial chromosome cdna clone
dc.creator.none.fl_str_mv Ávila-Pérez, Ginés
Park, Jun-Gyu
Nogales, Aitor
Almazán, Fernando
Martínez-Sobrido, Luis
author Ávila-Pérez, Ginés
author_facet Ávila-Pérez, Ginés
Park, Jun-Gyu
Nogales, Aitor
Almazán, Fernando
Martínez-Sobrido, Luis
author_role author
author2 Park, Jun-Gyu
Nogales, Aitor
Almazán, Fernando
Martínez-Sobrido, Luis
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Ministerio de Economía y Competitividad (España)
National Institutes of Health (US)
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Immunology and Infection
Issue 148
Arbovirus
Flavivirus
Zika virus
Reverse genetics
Full-length cDNA infectious clones
Bacterial artificial chromosomes
cDNA transfection
Virus rescue
topic Immunology and Infection
Issue 148
Arbovirus
Flavivirus
Zika virus
Reverse genetics
Full-length cDNA infectious clones
Bacterial artificial chromosomes
cDNA transfection
Virus rescue
description © 2019 Journal of Visualized Experiments.
publishDate 2019
dc.date.none.fl_str_mv 2019
2021
2021
2021
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/241180
url http://hdl.handle.net/10261/241180
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BFU2016-79127-R
http://dx.doi.org/10.3791/59537
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv MYJoVE Corporation
publisher.none.fl_str_mv MYJoVE Corporation
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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