Data/hora: 09/06/2024 05:53:58

Provedor de dados: | 102
País: | Brazil
Título: | Polyclonal antibodies to the coat protein of Apple stem grooving virus expressed in Escherichia coli: production and use in immunodiagnosis
Autores: | Nickel,Osmar; Targon,Maria L.P.N.; Fajardo,Thor V.M.; Machado,Marcos A.; Trivilin,Ana P.
Data: | 2004-10-01
Ano: | 2004
Palavras-chave: | Recombinant antigen; Latent virus; ELISA; Malus domestica.
Resumo: | The coat protein gene of Apple stem grooving virus (ASGV) was amplified by RT-PCR, cloned, sequenced and subcloned in the expression vector pMal-c2. This plasmid was used to transform Escherichia coli BL21c+ competent cells. The ASGV coat protein (cp) was expressed as a fusion protein containing a fragment of E. coli maltose binding protein (MBP). Bacterial cells were disrupted by sonication and the ASGVcp/MBP fusion protein was purified by amylose resin affinity chromatography. Polyclonal antibodies from rabbits immunized with the fusion protein gave specific reactions to ASGV from infected apple (Malus domestica) cv. Fuji Irradiada and Chenopodium quinoa at dilutions of up to 1:1,000 and 1:2,000, respectively, in plate trapped ELISA. The ASGVcp/MBP fusion protein reacted to a commercial antiserum against ASGV in immunoblotting assay. The IgG against ASGVcp/MBP performed favorably in specificity and sensitivity to the virus. This method represents an additional tool for the efficient ASGV-indexing of apple propagative and mother stock materials, and for use in support of biological and molecular techniques.
Tipo: | Info:eu-repo/semantics/report
Idioma: | Inglês
Identificador: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-41582004000500017
Editor: | Sociedade Brasileira de Fitopatologia
Relação: | 10.1590/S0100-41582004000500017
Formato: | text/html
Fonte: | Fitopatologia Brasileira  v.29 n.5 2004
Fonte: | info:eu-repo/semantics/openAccess