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

dc.creatorSilva Jr., J. V. J.
dc.creatorArenhart, S.
dc.creatorSantos, H. F.
dc.creatorAlmeida-Queiroz, S. R.
dc.creatorSilva, A. N. M. R.
dc.creatorTrevisol, I. M.
dc.creatorBertani, G. R.
dc.creatorGil, L. H. V. G.
dc.date2017-11-30T13:15:29Z
dc.date2017-11-30T13:15:29Z
dc.date2014
dc.date.accessioned2023-09-26T20:47:27Z
dc.date.available2023-09-26T20:47:27Z
dc.identifierSILVA JR., J. V. J. et al. Efficient assembly of full-length infectious clone of Brazilian IBDV isolate by homologous recombination in yeast. Brazilian Journal of Microbiology, v. 45, n. 4, p. 1555–1563, dez. 2014.
dc.identifier1678-4405
dc.identifierhttps://www.arca.fiocruz.br/handle/icict/23468
dc.identifier10.1590/S1517-83822014000400054
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8863948
dc.descriptionThe 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.
dc.formatapplication/pdf
dc.languageeng
dc.rightsopen access
dc.subjectVírus da Doença Infecciosa da Bursa
dc.subjectGenética Reversa
dc.subjectrecombinação homóloga baseada em fermento
dc.subjectinfectious bursal disease virus
dc.subjectreverse genetics
dc.subjectyeast-based homologous recombination
dc.subjectAnimais
dc.subjectEmbrião de Galinha
dc.subjectRecombinação Homóloga
dc.subjectVetores Genéticos
dc.subjectVírus da Doença Infecciosa da Bursa
dc.subjectgenética
dc.subjectGenética Reversa
dc.subjectmétodos
dc.subjectBrasil
dc.subjectCélulas Cultivadas
dc.subjectFibroblastos
dc.subjectvirologia
dc.subjectInstabilidade Genômica
dc.subjectTransfecção
dc.subjectVírus da Doença Infecciosa da Bursa
dc.subjectisolamento & purificação
dc.subjectVírus da Doença Infecciosa da Bursa
dc.subjectfisiologia
dc.subjectSaccharomyces cerevisiae
dc.subjectgenética
dc.subjectCultura de Vírus
dc.subjectReplicação Viral
dc.titleEfficient assembly of full-length infectious clone of Brazilian IBDV isolate by homologous recombination in yeast
dc.typeArticle


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