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| Radaelli,Paula; Fajardo,Thor Vinícius Martins; Nickel,Osmar; Eiras,Marcelo; Pio-Ribeiro,Gilvan. |
| The objective of this work was to produce and characterize specific antisera against Brazilian isolates of Grapevine leafroll-associated virus 2 (GLRaV-2) and Grapevine virus B (GVB), developed from expressed coat proteins (CPs) in Escherichia coli, and to test their possible use for the detection of these two viruses in diseased grapevines. The coat protein (CP) genes were RT-PCR-amplified, cloned and sequenced. The CP genes were subsequently subcloned, and the recombinant plasmids were used to transform E. coli cells and express the coat proteins. The recombinant coat proteins were purified, and their identities were confirmed by SDS-PAGE and Western blot and used for rabbit immunizations. Antisera raised against these proteins were able to recognize the... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Vitis; GLRaV-2; GVB; Indirect ELISA; Recombinant protein; Western blot. |
| Ano: 2008 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-204X2008001000020 |
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| NICKEL,OSMAR; FAJARDO,THOR V. M.; ARAGÃO,FRANCISCO J. L.; CHAGAS,CÉSAR M.; KUHN,GILMAR B.. |
| An isolate of Grapevine virus B (GVB), obtained by indexing Vitis labrusca and V. vinifera grapevines on the indicator LN33, was transmitted mechanically to several Nicotiana species. The virus was partially purified from N. cavicola and the coat protein estimated at 23 kDa by SDS-PAGE. In negatively stained leaf extracts of experimentally inoculated N. cavicola and N. occidentalis, flexuous particles with cross banding were observed, predominantly measuring 750-770 x 12 nm, with a modal length of 760 nm. Decoration indicated a clear, positive reaction against AS-GVB. In DAS-ELISA, GVB was detected in N. cavicola and grapevine extracts, and Western blots showed homologous and cross reaction of GVB and GVA antisera with GVB coat protein. Using specific... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: GVB; Vitis spp.; Vitivirus; Electron microscopy; ELISA; Western blot; GVA. |
| Ano: 2002 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-41582002000300007 |
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