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| Formato: | Artículo científico |
| Lenguaje: | en |
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Academia Brasileira de Ciências
2014
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| Acceso en línea: | https://www.redalyc.org/articulo.oa?id=32732907016 |
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- A two-plasmid strategy for engineering a dengue virus type 3 infectious clone from primary Brazilian isolate JEFFERSON J. S. SANTOS MARLI T. CORDEIRO GIOVANI R. BERTANI ERNESTO T. A. MARQUES LAURA H. V. G. GIL Multidisciplinaria (Ciencias Naturales y Exactas) dengue virus Reverse genetics infectious clone molecular cloning Dengue infections represent one of the most prevalent arthropod-borne diseases worldwide, causing a wide spectrum of clinical outcomes. Engineered infectious clone is an important tool to study Dengue virus (DENV) biology. Functional full-length cDNA clones have been constructed for many positivestrand RNA viruses and have provided valuable tools for studying the molecular mechanisms involved in viral genome replication, virion assembly, virus pathogenesis and vaccine development. We report herein the successful development of an infectious clone from a primary Brazilian isolate of dengue virus 3 (DENV3) of the genotype III. Using a two-plasmid strategy, DENV3 genome was divided in two parts and cloned separately into a yeast-bacteria shuttle vector. All plasmids were assembled in yeast by homologous recombination technique and a full-length template for transcription was obtained by in vitro ligation of the two parts of the genome. Transcript-derived DENV3 is infectious upon transfection into BHK-21 cells and in vitro characterization confirmed its identity. Growth kinetics of transcript-derived DENV3 was indistinguishable from wild type DENV3. This system is a powerful tool that will help shed light on molecular features of DENV biology, as the relationship of specific mutations and DENV pathogenesis. 2014 artículo científico 0001-3765 https://www.redalyc.org/articulo.oa?id=32732907016 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.4 Vol.86