Fitopatologia Brasileira (2004) 29, 215-219

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Manuela R. Barbieri, Murilo G. de Carvalho, Eunize M. Zambolim and F. Murilo Zerbini (2004)
Expressão em Escherichia coli da proteína capsidial do Watermelon mosaic virus e produção de anti-soro
[Expression in Escherichia coli of the capsid protein of Watermelon mosaic virus and production of specific antiserum]
Fitopatologia Brasileira 29 (2), 215-219
Abstract: A Watermelon mosaic virus (WMV)-specific, polyclonal antiserum was raised in rabbits by immunization with purified capsid protein expressed and purified from Escherichia coli cells. The cp gene was RT-PCR-amplified using specific oligonucleotides, and viral RNA extracted from concentrated particles as a template. The amplified fragment was cloned in pBLUESCRIPT KS+ and fully sequenced in order to confirm its identity and integrity. The fragment was subsequently subcloned into the pRSET-A expression vector. Recombinant plasmids were used to induce the expression of the capsid protein in E. coli BL21::DE3. The protein was purified by affinity chromatography using a Ni+-NTA resin, from E. coli total protein extract. The purified protein was quantified and used for the immunization of rabbits. The antiserum was tested for its sensitivity and specificity in Western blot, DAS-ELISA, immunodiffusion and indirect ELISA assays. All tests indicated that the antiserum raised from in vitro-expressed capsid protein is highly specific for the detection of WMV in plant extracts, with no interspecific, heterologous reactions being observed.
(The abstract is excluded from the Creative Commons licence and has been copied with permission by the publisher.)
(original language: Portuguese)
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Database assignments for author(s): F. Murilo Zerbini

Research topic(s) for pests/diseases/weeds:
molecular biology - genes

Pest and/or beneficial records:

Beneficial Pest/Disease/Weed Crop/Product Country Quarant.

Watermelon mosaic virus