Please use this identifier to cite or link to this item: http://www.alice.cnptia.embrapa.br/alice/handle/doc/952212
Research center of Embrapa/Collection: Embrapa Soja - Artigo em periódico indexado (ALICE)
Date Issued: 2013
Type of Material: Artigo em periódico indexado (ALICE)
Authors: CARVALHO, S. L. de
SILVA, F. N. da
ZANARDO, L. G.
ALMEIDA, A. M. R.
ZERBINI, F. M.
CARVALHO, C. M.
Additional Information: 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.
Title: Production of polyclonal antiserum against Cowpea mild mottle virus coat protein and its application in virus detection.
Publisher: Tropical Plant Patology, Brasília, DF, v. 38, n. 1, p. 49-54, Jan.-Feb. 2013.
Language: en
Description: 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.
Thesagro: Doença de planta
Vírus
Soja
NAL Thesaurus: Plant diseases and disorders
Cowpea mild mottle virus
Soybeans
Data Created: 2013-03-05
ISSN: 1982-5676
Appears in Collections:Artigo em periódico indexado (CNPSO)

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