Efficient assembly of full-length infectious clone of Brazilian IBDV isolate by homologous recombination in yeast

Detalhes bibliográficos
Autor(a) principal: Silva Jr.,J.V.J.
Data de Publicação: 2014
Outros Autores: Arenhart,S., Santos,H.F., Almeida-Queiroz,S.R., Silva,A.N.M.R., Trevisol,I.M., Bertani,G.R., Gil,L.H.V.G.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Brazilian Journal of Microbiology
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000400054
Resumo: The Infectious Bursal Disease Virus (IBDV) causes immunosuppression in young chickens. Advances in molecular virology and vaccines for IBDV have been achieved by viral reverse genetics (VRG). VRG for IBDV has undergone changes over time, however all strategies used to generate particles of IBDV involves multiple rounds of amplification and need of in vitro ligation and restriction sites. The aim of this research was to build the world's first VRG for IBDV by yeast-based homologous recombination; a more efficient, robust and simple process than cloning by in vitro ligation. The wild type IBDV (Wt-IBDV-Br) was isolated in Brazil and had its genome cloned in pJG-CMV-HDR vector by yeast-based homologous recombination. The clones were transfected into chicken embryo fibroblasts and the recovered virus (IC-IBDV-Br) showed genetic stability and similar phenotype to Wt-IBDV-Br, which were observed by nucleotide sequence, focus size/morphology and replication kinetics, respectively. Thus, IBDV reverse genetics by yeast-based homologous recombination provides tools to IBDV understanding and vaccines/viral vectors development.
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spelling Efficient assembly of full-length infectious clone of Brazilian IBDV isolate by homologous recombination in yeastinfectious bursal disease virusreverse geneticsyeast-based homologous recombinationThe Infectious Bursal Disease Virus (IBDV) causes immunosuppression in young chickens. Advances in molecular virology and vaccines for IBDV have been achieved by viral reverse genetics (VRG). VRG for IBDV has undergone changes over time, however all strategies used to generate particles of IBDV involves multiple rounds of amplification and need of in vitro ligation and restriction sites. The aim of this research was to build the world's first VRG for IBDV by yeast-based homologous recombination; a more efficient, robust and simple process than cloning by in vitro ligation. The wild type IBDV (Wt-IBDV-Br) was isolated in Brazil and had its genome cloned in pJG-CMV-HDR vector by yeast-based homologous recombination. The clones were transfected into chicken embryo fibroblasts and the recovered virus (IC-IBDV-Br) showed genetic stability and similar phenotype to Wt-IBDV-Br, which were observed by nucleotide sequence, focus size/morphology and replication kinetics, respectively. Thus, IBDV reverse genetics by yeast-based homologous recombination provides tools to IBDV understanding and vaccines/viral vectors development.Sociedade Brasileira de Microbiologia2014-12-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000400054Brazilian Journal of Microbiology v.45 n.4 2014reponame:Brazilian Journal of Microbiologyinstname:Sociedade Brasileira de Microbiologia (SBM)instacron:SBM10.1590/S1517-83822014000400054info:eu-repo/semantics/openAccessSilva Jr.,J.V.J.Arenhart,S.Santos,H.F.Almeida-Queiroz,S.R.Silva,A.N.M.R.Trevisol,I.M.Bertani,G.R.Gil,L.H.V.G.eng2015-02-13T00:00:00Zoai:scielo:S1517-83822014000400054Revistahttps://www.scielo.br/j/bjm/ONGhttps://old.scielo.br/oai/scielo-oai.phpbjm@sbmicrobiologia.org.br||mbmartin@usp.br1678-44051517-8382opendoar:2015-02-13T00:00Brazilian Journal of Microbiology - Sociedade Brasileira de Microbiologia (SBM)false
dc.title.none.fl_str_mv Efficient assembly of full-length infectious clone of Brazilian IBDV isolate by homologous recombination in yeast
title Efficient assembly of full-length infectious clone of Brazilian IBDV isolate by homologous recombination in yeast
spellingShingle Efficient assembly of full-length infectious clone of Brazilian IBDV isolate by homologous recombination in yeast
Silva Jr.,J.V.J.
infectious bursal disease virus
reverse genetics
yeast-based homologous recombination
title_short Efficient assembly of full-length infectious clone of Brazilian IBDV isolate by homologous recombination in yeast
title_full Efficient assembly of full-length infectious clone of Brazilian IBDV isolate by homologous recombination in yeast
title_fullStr Efficient assembly of full-length infectious clone of Brazilian IBDV isolate by homologous recombination in yeast
title_full_unstemmed Efficient assembly of full-length infectious clone of Brazilian IBDV isolate by homologous recombination in yeast
title_sort Efficient assembly of full-length infectious clone of Brazilian IBDV isolate by homologous recombination in yeast
author Silva Jr.,J.V.J.
author_facet Silva Jr.,J.V.J.
Arenhart,S.
Santos,H.F.
Almeida-Queiroz,S.R.
Silva,A.N.M.R.
Trevisol,I.M.
Bertani,G.R.
Gil,L.H.V.G.
author_role author
author2 Arenhart,S.
Santos,H.F.
Almeida-Queiroz,S.R.
Silva,A.N.M.R.
Trevisol,I.M.
Bertani,G.R.
Gil,L.H.V.G.
author2_role author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Silva Jr.,J.V.J.
Arenhart,S.
Santos,H.F.
Almeida-Queiroz,S.R.
Silva,A.N.M.R.
Trevisol,I.M.
Bertani,G.R.
Gil,L.H.V.G.
dc.subject.por.fl_str_mv infectious bursal disease virus
reverse genetics
yeast-based homologous recombination
topic infectious bursal disease virus
reverse genetics
yeast-based homologous recombination
description The Infectious Bursal Disease Virus (IBDV) causes immunosuppression in young chickens. Advances in molecular virology and vaccines for IBDV have been achieved by viral reverse genetics (VRG). VRG for IBDV has undergone changes over time, however all strategies used to generate particles of IBDV involves multiple rounds of amplification and need of in vitro ligation and restriction sites. The aim of this research was to build the world's first VRG for IBDV by yeast-based homologous recombination; a more efficient, robust and simple process than cloning by in vitro ligation. The wild type IBDV (Wt-IBDV-Br) was isolated in Brazil and had its genome cloned in pJG-CMV-HDR vector by yeast-based homologous recombination. The clones were transfected into chicken embryo fibroblasts and the recovered virus (IC-IBDV-Br) showed genetic stability and similar phenotype to Wt-IBDV-Br, which were observed by nucleotide sequence, focus size/morphology and replication kinetics, respectively. Thus, IBDV reverse genetics by yeast-based homologous recombination provides tools to IBDV understanding and vaccines/viral vectors development.
publishDate 2014
dc.date.none.fl_str_mv 2014-12-01
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000400054
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000400054
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1590/S1517-83822014000400054
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv text/html
dc.publisher.none.fl_str_mv Sociedade Brasileira de Microbiologia
publisher.none.fl_str_mv Sociedade Brasileira de Microbiologia
dc.source.none.fl_str_mv Brazilian Journal of Microbiology v.45 n.4 2014
reponame:Brazilian Journal of Microbiology
instname:Sociedade Brasileira de Microbiologia (SBM)
instacron:SBM
instname_str Sociedade Brasileira de Microbiologia (SBM)
instacron_str SBM
institution SBM
reponame_str Brazilian Journal of Microbiology
collection Brazilian Journal of Microbiology
repository.name.fl_str_mv Brazilian Journal of Microbiology - Sociedade Brasileira de Microbiologia (SBM)
repository.mail.fl_str_mv bjm@sbmicrobiologia.org.br||mbmartin@usp.br
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