Data/hora: 01/06/2024 14:39:47

Provedor de dados: 	42	
País: 	Brazil	
Título: 	Construction of yellow fever virus subgenomic replicons by yeast-based homologous recombination cloning technique	
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.	
Data: 	2013-03-01	
Ano: 	2013	
Palavras-chave: 	Cloning technique; Homologous recombination; Replicon; Reporter gene; Yellow fever virus.	
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.	
Tipo: 	Info:eu-repo/semantics/article	
Idioma: 	Inglês	
Identificador: 	http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652013000100159	
Editor: 	Academia Brasileira de Ciências	
Relação: 	10.1590/S0001-37652013005000008	
Formato: 	text/html	
Fonte: 	Anais da Academia Brasileira de Ciências  v.85 n.1 2013	
Fonte: 	info:eu-repo/semantics/openAccess