| Registro completo |
| Provedor de dados: |
Anais da ABC (AABC)
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| País: |
Brazil
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| Título: |
Construction of yellow fever virus subgenomic replicons by yeast-based homologous recombination cloning technique
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| Autores: |
Queiroz,Sabrina R.A.
Silva,Andréa N.M.R.
Santos,Jefferson J.S.
Marques Jr,Ernesto T.A.
Bertani,Giovani R.
Gil,Laura H.V.G.
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| Data: |
2013-03-01
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| Ano: |
2013
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| Palavras-chave: |
Cloning technique
Homologous recombination
Replicon
Reporter gene
Yellow fever virus
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| Resumo: |
RNA replicon derived from Flavivirus genome is a valuable tool for studying viral replication independent of virion assembly and maturation, besides being a great potencial for heterologous gene expression. In this study we described the construction of subgenomic replicons of yellow fever virus by yeast-based homologous recombination technique. The plasmid containing the yellow fever 17D strain replicon (pBSC-repYFV-17D), previously characterized, was handled to heterologous expression of the green fluorescent protein (repYFV-17D-GFP) and firefly luciferase (repYFV-17D-Luc) reporter genes. Both replicons were constructed by homologous recombination between the linearized vector pBSC-repYFV-17D and the PCR product containing homologous 25 nucleotides ends incorporated into PCR primers. The genomic organization of these constructs is similar to repYFV-17D, but with insertion of the reporter gene between the remaining 63 N-terminal nucleotides of the capsid protein and 72 C-terminal nucleotides of the E protein. The replicons repYFV-17D-GFP and repYFV-17D-Luc showed efficient replication and expression of the reporter genes. The yeast-based homologous recombination technique used in this study proved to be applicable for manipulation of the yellow fever virus genome in order to construct subgenomic replicons.
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| Tipo: |
Info:eu-repo/semantics/article
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| Idioma: |
Inglês
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| Identificador: |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652013000100159
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| Editor: |
Academia Brasileira de Ciências
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| Relação: |
10.1590/S0001-37652013005000008
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| Formato: |
text/html
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| Fonte: |
Anais da Academia Brasileira de Ciências v.85 n.1 2013
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| Direitos: |
info:eu-repo/semantics/openAccess
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