Data/hora: 09/06/2024 05:43:02

Provedor de dados: | 102
País: | Brazil
Título: | Expression of Grapevine leafroll-associated virus 3 coat protein gene in Escherichia coli and production of polyclonal antibodies
Autores: | Fajardo,Thor V.M.; Barros,Danielle R.; Nickel,Osmar; Kuhn,Gilmar B.; Zerbini,F. Murilo.
Data: | 2007-12-01
Ano: | 2007
Palavras-chave: | Grapevine; GLRaV-3; Recombinant protein; Serology; Antibodies.
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.
Tipo: | Info:eu-repo/semantics/other
Idioma: | Inglês
Identificador: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-41582007000600007
Editor: | Sociedade Brasileira de Fitopatologia
Relação: | 10.1590/S0100-41582007000600007
Formato: | text/html
Fonte: | Fitopatologia Brasileira  v.32 n.6 2007
Fonte: | info:eu-repo/semantics/openAccess