Efficient assembly of full-length infectious clone of Brazilian IBDV isolate by homologous recombination in yeast.
Autor(a) principal: | |
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Data de Publicação: | 2014 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) |
Texto Completo: | http://www.alice.cnptia.embrapa.br/alice/handle/doc/1026478 |
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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Efficient assembly of full-length infectious clone of Brazilian IBDV isolate by homologous recombination in yeast.IBVDVirologia molecularFrangoDoença de GumboroGenética molecularChickensInfectious bursal disease virusAvibirnavirusVertebrate virusesMolecular geneticsThe 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.JOSÉ VALTER JOAQUIM SILVA JÚNIOR, FIOCRUZ; SANDRA ARENHART, FIOCRUZ; HELTON FERNANDES DOS SANTOS, UFRGS/ICBS; SABRINA RIBEIRO DE ALMEIDA-QUEIROZ, FIOCRUZ; ANDRÉA NAZARÉ MONTEIRO RANGEL DA SILVA, FIOCRUZ; IARA MARIA TREVISOL, CNPSA; GIOVANI ROTA, UFPE/LIKA; LAURA HELENA VEGA GONZALES GIL, FIOCRUZ.SILVA JÚNIOR, J. V. J.ARENHART, S.SANTOS, H. F. dosALMEIDA-QUEIROZ, S. R.SILVA, A. N. M. R.TREVISOL, I. M.BERTANI, G. R.GIL, L. H. V. G.2018-01-26T23:42:11Z2018-01-26T23:42:11Z2015-10-1520142018-01-26T23:42:11Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleBrazilian Journal of Microbiology, v. 45, n. 4, p. 1555-1563, 2014.http://www.alice.cnptia.embrapa.br/alice/handle/doc/1026478enginfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice)instname:Empresa Brasileira de Pesquisa Agropecuária (Embrapa)instacron:EMBRAPA2018-01-26T23:42:17Zoai:www.alice.cnptia.embrapa.br:doc/1026478Repositório InstitucionalPUBhttps://www.alice.cnptia.embrapa.br/oai/requestopendoar:21542018-01-26T23:42:17falseRepositório InstitucionalPUBhttps://www.alice.cnptia.embrapa.br/oai/requestcg-riaa@embrapa.bropendoar:21542018-01-26T23:42:17Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa)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 JÚNIOR, J. V. J. IBVD Virologia molecular Frango Doença de Gumboro Genética molecular Chickens Infectious bursal disease virus Avibirnavirus Vertebrate viruses Molecular genetics |
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 JÚNIOR, J. V. J. |
author_facet |
SILVA JÚNIOR, J. V. J. ARENHART, S. SANTOS, H. F. dos 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. dos 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.none.fl_str_mv |
JOSÉ VALTER JOAQUIM SILVA JÚNIOR, FIOCRUZ; SANDRA ARENHART, FIOCRUZ; HELTON FERNANDES DOS SANTOS, UFRGS/ICBS; SABRINA RIBEIRO DE ALMEIDA-QUEIROZ, FIOCRUZ; ANDRÉA NAZARÉ MONTEIRO RANGEL DA SILVA, FIOCRUZ; IARA MARIA TREVISOL, CNPSA; GIOVANI ROTA, UFPE/LIKA; LAURA HELENA VEGA GONZALES GIL, FIOCRUZ. |
dc.contributor.author.fl_str_mv |
SILVA JÚNIOR, J. V. J. ARENHART, S. SANTOS, H. F. dos 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 |
IBVD Virologia molecular Frango Doença de Gumboro Genética molecular Chickens Infectious bursal disease virus Avibirnavirus Vertebrate viruses Molecular genetics |
topic |
IBVD Virologia molecular Frango Doença de Gumboro Genética molecular Chickens Infectious bursal disease virus Avibirnavirus Vertebrate viruses Molecular genetics |
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 2015-10-15 2018-01-26T23:42:11Z 2018-01-26T23:42:11Z 2018-01-26T23:42:11Z |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/publishedVersion info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
Brazilian Journal of Microbiology, v. 45, n. 4, p. 1555-1563, 2014. http://www.alice.cnptia.embrapa.br/alice/handle/doc/1026478 |
identifier_str_mv |
Brazilian Journal of Microbiology, v. 45, n. 4, p. 1555-1563, 2014. |
url |
http://www.alice.cnptia.embrapa.br/alice/handle/doc/1026478 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) instname:Empresa Brasileira de Pesquisa Agropecuária (Embrapa) instacron:EMBRAPA |
instname_str |
Empresa Brasileira de Pesquisa Agropecuária (Embrapa) |
instacron_str |
EMBRAPA |
institution |
EMBRAPA |
reponame_str |
Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) |
collection |
Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) |
repository.name.fl_str_mv |
Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa) |
repository.mail.fl_str_mv |
cg-riaa@embrapa.br |
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1794503448767496192 |