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| Kanehira,Katsushi; Uchida,Yuko; Saito,Takehiko. |
| Background: Avian influenza viruses (AIVs) are influenza A viruses which are isolated from domestic and wild birds. AIVs that include highly pathogenic avian influenza viruses (HPAIVs) are a major concern to the poultry industry because they cause outbreaks in poultry with extraordinarily high lethality. In addition, AIVs threaten human health by occasional zoonotic infection of humans from birds. Tools to visualize AIV-infected cells would facilitate the development of diagnostic tests and preventative methods to reduce the spread of AIVs. In this study, a self-assembling split-green fluorescent protein (split-GFP) system, combined with influenza virus reverse genetics was used to construct a visualization method for influenza virus-infected cells.... |
| Tipo: Journal article |
Palavras-chave: Avian influenza virus; Reverse genetics; Split-GFP. |
| Ano: 2016 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000100010 |
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| Arenhart,Sandra; Silva Junior,José Valter Joaquim; Flores,Eduardo Furtado; Weiblen,Rudi; Gil,Laura Helena Vega Gonzales. |
| Abstract The open reading frame of a Brazilian bovine viral diarrhea virus (BVDV) strain, IBSP4ncp, was recombined with the untranslated regions of the reference NADL strain by homologous recombination in Saccharomyces cerevisiae, resulting in chimeric full-length cDNA clones of BVDV (chi-NADL/IBSP4ncp#2 and chi-NADL/IBSP4ncp#3). The recombinant clones were successfully recovered, resulting in viable viruses, having the kinetics of replication, focus size, and morphology similar to those of the parental virus, IBSP4ncp. In addition, the chimeric viruses remained stable for at least 10 passages in cell culture, maintaining their replication efficiency unaltered. Nucleotide sequencing revealed a few point mutations; nevertheless, the phenotype of the rescued... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: BVDV; Pestivirus; Reverse genetics; Infectious clone; Yeast homologous recombination. |
| Ano: 2016 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822016000400993 |
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| 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.. |
| 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... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Infectious bursal disease virus; Reverse genetics; Yeast-based homologous recombination. |
| Ano: 2014 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000400054 |
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| Berenschot,Amanda S.; Zucchi,Maria I.; Tulmann-Neto,Augusto; Quecini,Vera. |
| Traditionally, mutagenesis has been used to introduce novel genetic variability in ornamental crops. More recently, it has become a powerful tool in gene discovery and functional analyses in reverse genetics approaches. The present work aimed to compare the efficiency of physical and chemical agents in generating mutant populations of petunia. We have indirectly evaluated the genomic damage by analyzing developmental characteristics of the plantlets derived from treated seeds employing gamma radiation at 0, 20, 40, 60, 80 and 100 Gy and the alkylating agent ethyl-methanesulfonate (EMS) at 0, 0.05, 0.1, 0.15, 0.2 and 0.25% (v/v). Gamma rays and EMS caused developmental defects and decreased seedling viability in plants obtained from the mutagenized seeds.... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: EMS; Gamma irradiation; Genomic damage; Mutagenesis; Reverse genetics. |
| Ano: 2008 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1677-04202008000200002 |
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