title: Efficient assembly of full-length infectious clone of Brazilian IBDV isolate by homologous recombination in yeast.
abstract: 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.
authors.affiliation: Departamento de Virologia e Terapia Experimental Centro de Pesquisas Aggeu Magalhães Fundação Oswaldo Cruz RecifePE Brazil Departamento de Virologia e Terapia Experimental, Centro de Pesquisas Aggeu Magalhães, Fundação Oswaldo Cruz, Recife, PE, Brazil.
authors.forename: J V J
authors.initials: JV
authors.lastname: Silva
authors.affiliation: Departamento de Virologia e Terapia Experimental Centro de Pesquisas Aggeu Magalhães Fundação Oswaldo Cruz RecifePE Brazil Departamento de Virologia e Terapia Experimental, Centro de Pesquisas Aggeu Magalhães, Fundação Oswaldo Cruz, Recife, PE, Brazil. ; Setor de Virologia, Departamento de Medicina Veterinária Preventiva Centro de Ciências Rurais Universidade Federal de Santa Maria Santa MariaRS Brazil Setor de Virologia, Departamento de Medicina Veterinária Preventiva, Centro de Ciências Rurais, Universidade Federal de Santa Maria, Santa Maria, RS, Brazil.
authors.forename: S
authors.initials: S
authors.lastname: Arenhart
authors.affiliation: Laboratório de Virologia, Departamento de Microbiologia Imunologia e Parasitologia, Instituto de Ciências Básicas da Saúde Universidade Federal do Rio Grande do Sul Porto AlegreRS Brazil Laboratório de Virologia, Departamento de Microbiologia, Imunologia e Parasitologia, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil.
authors.forename: H F
authors.initials: HF
authors.lastname: Santos
authors.affiliation: Departamento de Virologia e Terapia Experimental Centro de Pesquisas Aggeu Magalhães Fundação Oswaldo Cruz RecifePE Brazil Departamento de Virologia e Terapia Experimental, Centro de Pesquisas Aggeu Magalhães, Fundação Oswaldo Cruz, Recife, PE, Brazil.
authors.forename: S R
authors.initials: SR
authors.lastname: Almeida-Queiroz
authors.affiliation: Departamento de Virologia e Terapia Experimental Centro de Pesquisas Aggeu Magalhães Fundação Oswaldo Cruz RecifePE Brazil Departamento de Virologia e Terapia Experimental, Centro de Pesquisas Aggeu Magalhães, Fundação Oswaldo Cruz, Recife, PE, Brazil.
authors.forename: A N M R
authors.initials: AN
authors.lastname: Silva
authors.affiliation: Embrapa Suínos e Aves ConcórdiaSC Brazil Embrapa Suínos e Aves, Concórdia, SC, Brazil.
authors.forename: I M
authors.initials: IM
authors.lastname: Trevisol
authors.affiliation: Departamento de Bioquímica e Laboratório de Imunopatologia Keiso Asami Universidade Federal de Pernambuco RecifePE Brazil Departamento de Bioquímica e Laboratório de Imunopatologia Keiso Asami, Universidade Federal de Pernambuco, Recife, PE, Brazil.
authors.forename: G R
authors.initials: GR
authors.lastname: Bertani
authors.affiliation: Departamento de Virologia e Terapia Experimental Centro de Pesquisas Aggeu Magalhães Fundação Oswaldo Cruz RecifePE Brazil Departamento de Virologia e Terapia Experimental, Centro de Pesquisas Aggeu Magalhães, Fundação Oswaldo Cruz, Recife, PE, Brazil.
authors.forename: L H V G
authors.initials: LH
authors.lastname: Gil
mesh_terms: D000818:Animals; D001938:Brazil; D002478:Cells, Cultured; D002642:Chick Embryo; D005347:Fibroblasts / Q000821:virology; D005822:Genetic Vectors; D042822:Genomic Instability; D059765:Homologous Recombination; D007243:Infectious bursal disease virus / Q000235:genetics* / Q000302:isolation & purification / Q000502:physiology; D059386:Reverse Genetics / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014162:Transfection; D014776:Virus Cultivation; D014779:Virus Replication
keywords: infectious bursal disease virus; reverse genetics; yeast-based homologous recombination