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| Format: | Artículo científico |
| Language: | en |
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Academia Brasileira de Ciências
2017
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| Online Access: | https://www.redalyc.org/articulo.oa?id=32753602015 |
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| _version_ | 1866815446500507648 |
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| author | TELISSA C. KASSAR |
| author_facet | TELISSA C. KASSAR |
| contents | Construction and characterization of a recombinant yellow fever virus stably expressing Gaussia luciferase TELISSA C. KASSAR TEREZA MAGALHÃES JOSÉ V.J.S. JÚNIOR AMANDA G.O. CARVALHO ANDRÉA N.M.R. DA SILVA SABRINA R.A. QUEIROZ GIOVANI R. BERTANI LAURA H.V.G. GIL Multidisciplinaria (Ciencias Naturales y Exactas) reporter gene Gaussia luciferase yellow fever virus homologous recombination in yeast Yellow fever is an arthropod-borne viral disease that still poses high public health concerns, despite the availability of an effective vaccine. The development of recombinant viruses is of utmost importance for several types of studies, such as those aimed to dissect virus-host interactions and to search for novel antiviral strategies. Moreover, recombinant viruses expressing reporter genes may greatly facilitate these studies. Here, we report the construction of a recombinant yellow fever virus (YFV) expressing Gaussia luciferase (GLuc) (YFV-GLuc). We show, through RT-PCR, sequencing and measurement of GLuc activity, that stability of the heterologous gene was maintained after six passages. Furthermore, a direct association between GLuc expression and viral replication was observed (r 2 =0.9967), indicating that measurement of GLuc activity may be used to assess viral replication in different applications. In addition, we evaluated the use of the recombinant virus in an antiviral assay with recombinant human alfa-2b interferon. A 60% inhibition of GLuc expression was observed in cells infected with YFV-GLuc and incubated with IFN alfa-2b. Previously tested on YFV inhibition by plaque assays indicated a similar fold-decrease in viral replication. These results are valuable as they show the stability of YFV-GLuc and one of several possible applications of this construct. 2017 artículo científico 0001-3765 https://www.redalyc.org/articulo.oa?id=32753602015 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.3 Vol.89 |
| format | Artículo científico |
| id | redalyc_32753602015 |
| language | en |
| publishDate | 2017 |
| publisher | Academia Brasileira de Ciências |
| spellingShingle | Construction and characterization of a recombinant yellow fever virus stably expressing Gaussia luciferase TELISSA C. KASSAR Multidisciplinaria (Ciencias Naturales y Exactas) reporter gene Gaussia luciferase yellow fever virus homologous recombination in yeast Construction and characterization of a recombinant yellow fever virus stably expressing Gaussia luciferase TELISSA C. KASSAR TEREZA MAGALHÃES JOSÉ V.J.S. JÚNIOR AMANDA G.O. CARVALHO ANDRÉA N.M.R. DA SILVA SABRINA R.A. QUEIROZ GIOVANI R. BERTANI LAURA H.V.G. GIL Multidisciplinaria (Ciencias Naturales y Exactas) reporter gene Gaussia luciferase yellow fever virus homologous recombination in yeast Yellow fever is an arthropod-borne viral disease that still poses high public health concerns, despite the availability of an effective vaccine. The development of recombinant viruses is of utmost importance for several types of studies, such as those aimed to dissect virus-host interactions and to search for novel antiviral strategies. Moreover, recombinant viruses expressing reporter genes may greatly facilitate these studies. Here, we report the construction of a recombinant yellow fever virus (YFV) expressing Gaussia luciferase (GLuc) (YFV-GLuc). We show, through RT-PCR, sequencing and measurement of GLuc activity, that stability of the heterologous gene was maintained after six passages. Furthermore, a direct association between GLuc expression and viral replication was observed (r 2 =0.9967), indicating that measurement of GLuc activity may be used to assess viral replication in different applications. In addition, we evaluated the use of the recombinant virus in an antiviral assay with recombinant human alfa-2b interferon. A 60% inhibition of GLuc expression was observed in cells infected with YFV-GLuc and incubated with IFN alfa-2b. Previously tested on YFV inhibition by plaque assays indicated a similar fold-decrease in viral replication. These results are valuable as they show the stability of YFV-GLuc and one of several possible applications of this construct. 2017 artículo científico 0001-3765 https://www.redalyc.org/articulo.oa?id=32753602015 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.3 Vol.89 |
| title | Construction and characterization of a recombinant yellow fever virus stably expressing Gaussia luciferase |
| topic | Multidisciplinaria (Ciencias Naturales y Exactas) reporter gene Gaussia luciferase yellow fever virus homologous recombination in yeast |
| url | https://www.redalyc.org/articulo.oa?id=32753602015 |