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Registro Completo |
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Biblioteca(s): |
Embrapa Soja. |
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Data corrente: |
05/03/2013 |
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Data da última atualização: |
25/03/2013 |
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Autoria: |
CARVALHO, S. L. de; SILVA, F. N. da; ZANARDO, L. G.; ALMEIDA, A. M. R.; ZERBINI, F. M.; CARVALHO, C. M. |
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Afiliação: |
SILVIA L. DE CARVALHO, UFV; FÁBIO N. DA SILVA, UFV; LARISSA G. ZANARDO, UFV; ALVARO MANUEL RODRIGUES ALMEIDA, CNPSO; F. MURILO ZERBINI, UFV; CLAUDINE M. CARVALHO, UFV. |
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Título: |
Production of polyclonal antiserum against Cowpea mild mottle virus coat protein and its application in virus detection. |
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Ano de publicação: |
2013 |
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Fonte/Imprenta: |
Tropical Plant Patology, Brasília, DF, v. 38, n. 1, p. 49-54, Jan.-Feb. 2013. |
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ISSN: |
1982-5676 |
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Idioma: |
Inglês |
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Conteúdo: |
Cowpea mild mottle virus (CpMMV), the causal agent of stem necrosis disease, has drawn attention of soybean producers in recent years due to yield losses in the main producing regions of Brazil. Serological methods for viral detection require the use of an antiserum of good quality to achieve specificity and sensitivity. The entire coat protein gene of a Brazilian CpMMV isolate was cloned into a bacterial expression vector and transformed into Escherichia coli BL21::DE3 for in vitro expression. The coat protein, fused to a His-tag, was purified under denaturing conditions by affinity chromatography using a Ni-NTA resin. After renaturation, the integrity and identity of the purified recombinant protein was confirmed by SDS-Page and MALDI-ToF/ToF mass spectrometer analyses. A rabbit was immunized with increasing amounts of the recombinant protein. The specificity and sensitivity of the antiserum was demonstrated by Western blot and indirect ELISA assays. The polyclonal antisera raised against recombinant coat protein proved to be a reliable tool for CpMMV detection. |
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Thesagro: |
Doença de planta; Soja; Vírus. |
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Thesaurus Nal: |
Cowpea mild mottle virus; Plant diseases and disorders; Soybeans. |
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Categoria do assunto: |
F Plantas e Produtos de Origem Vegetal |
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URL: |
https://ainfo.cnptia.embrapa.br/digital/bitstream/item/77969/1/ID-34307.pdf
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Marc: |
LEADER 01905naa a2200265 a 4500
001 1952212
005 2013-03-25
008 2013 bl uuuu u00u1 u #d
022 $a1982-5676
100 1 $aCARVALHO, S. L. de
245 $aProduction of polyclonal antiserum against Cowpea mild mottle virus coat protein and its application in virus detection.$h[electronic resource]
260 $c2013
520 $aCowpea mild mottle virus (CpMMV), the causal agent of stem necrosis disease, has drawn attention of soybean producers in recent years due to yield losses in the main producing regions of Brazil. Serological methods for viral detection require the use of an antiserum of good quality to achieve specificity and sensitivity. The entire coat protein gene of a Brazilian CpMMV isolate was cloned into a bacterial expression vector and transformed into Escherichia coli BL21::DE3 for in vitro expression. The coat protein, fused to a His-tag, was purified under denaturing conditions by affinity chromatography using a Ni-NTA resin. After renaturation, the integrity and identity of the purified recombinant protein was confirmed by SDS-Page and MALDI-ToF/ToF mass spectrometer analyses. A rabbit was immunized with increasing amounts of the recombinant protein. The specificity and sensitivity of the antiserum was demonstrated by Western blot and indirect ELISA assays. The polyclonal antisera raised against recombinant coat protein proved to be a reliable tool for CpMMV detection.
650 $aCowpea mild mottle virus
650 $aPlant diseases and disorders
650 $aSoybeans
650 $aDoença de planta
650 $aSoja
650 $aVírus
700 1 $aSILVA, F. N. da
700 1 $aZANARDO, L. G.
700 1 $aALMEIDA, A. M. R.
700 1 $aZERBINI, F. M.
700 1 $aCARVALHO, C. M.
773 $tTropical Plant Patology, Brasília, DF$gv. 38, n. 1, p. 49-54, Jan.-Feb. 2013.
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Registro original: |
Embrapa Soja (CNPSO) |
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| 4. |  | ZORATO, M. F.; FRANÇA-NETO, J. B.; ASTAFEIEF, N. C.; TAKEDA, C. Nova interpretação na metodologia do teste de tetrazólio. Informativo ABRATES, Londrina, v.11, n.2, p.216, set. 2001. Numero especial, ref. 338. Edicao de Resumos do XII Congresso Brasileiro de Sementes, 17 a 20 de Setembro de 2001, Curitiba, PR.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Soja. |
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| 5. | | ZORATO, M. F.; FRANÇA-NETO, J. B.; ASTAFEIEF, N. C.; TAKEDA, C. Nova interpretação na metodologia do teste de tetrazólio. In: CONGRESSO BRASILEIRO DE SOJA, 2.; MERCOSOJA 2002, 2002, Foz do Iguaçu. Perspectivas do agronegócio da soja: resumos. Londrina: Embrapa Soja, 2002. p. 369. (Embrapa Soja. Documentos, 181). Organizado por Odilon Ferreira Saraiva, Clara Beatriz Hoffmann-Campo.| Tipo: Resumo em Anais de Congresso |
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