| Registro completo |
| Provedor de dados: |
Trop. Plant Pathol.
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| País: |
Brazil
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| Título: |
Expression of Grapevine leafroll-associated virus 3 coat protein gene in Escherichia coli and production of polyclonal antibodies
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| Autores: |
Fajardo,Thor V.M.
Barros,Danielle R.
Nickel,Osmar
Kuhn,Gilmar B.
Zerbini,F. Murilo
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| Data: |
2007-12-01
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| Ano: |
2007
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| Palavras-chave: |
Grapevine
GLRaV-3
Recombinant protein
Serology
Antibodies
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| Resumo: |
Grapevine leafroll-associated virus 3 (GLRaV-3), the main viral species of the grapevine leafroll complex, causes yield and quality reduction in grapes (Vitis spp.). The coat protein gene was RT-PCR-amplified from total RNA extracted from infected grapevine leaves and the amplified fragment was cloned and completely sequenced. The fragment was subsequently subcloned into the pRSET-C expression vector. The recombinant plasmid was used to transform Escherichia coli BL21:DE3 and express the capsid protein. The coat protein, fused to a 6 His-tag, was purified by affinity chromatography using an Ni-NTA resin. The identity of the purified protein was confirmed by SDS-PAGE and Western blot. The in vitro-expressed protein was quantified and used for rabbit immunizations. The antiserum was shown to be sensitive and specific for the detection of GLRaV-3 in grapevine extracts in Western blot and DAS-ELISA assays, with no unspecific or heterologous reactions against other non-serologically related viruses being observed.
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| Tipo: |
Info:eu-repo/semantics/other
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| Idioma: |
Inglês
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| Identificador: |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-41582007000600007
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| Editor: |
Sociedade Brasileira de Fitopatologia
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| Relação: |
10.1590/S0100-41582007000600007
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| Formato: |
text/html
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| Fonte: |
Fitopatologia Brasileira v.32 n.6 2007
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| Direitos: |
info:eu-repo/semantics/openAccess
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