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| Format: | Artículo científico |
| Language: | en |
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Academia Brasileira de Ciências
2013
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| Online Access: | https://www.redalyc.org/articulo.oa?id=32725624012 |
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Table of Contents:
- Construction of yellow fever virus subgenomic replicons by yeast-based homologous recombination cloning technique SABRINA R.A. QUEIROZ ANDRÉA N.M.R. SILVA JEFFERSON J.S. SANTOS ERNESTO T.A. MARQUES Jr GIOVANI R. BERTANI LAURA H.V.G. GIL Multidisciplinaria (Ciencias Naturales y Exactas) replicon reporter gene Cloning technique yellow fever virus homologous recombination 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. 2013 artículo científico 0001-3765 https://www.redalyc.org/articulo.oa?id=32725624012 en http://www.redalyc.org/revista.oa?id=327 Anais da Academia Brasileira de Ciências application/pdf Academia Brasileira de Ciências Anais da Academia Brasileira de Ciências (Brasil) Num.1 Vol.85