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Polyclonal Antibodies to the Coat Protein of Carnation etched ring virus Expressed in Bacterial System: Production and Use in Immunodiagnosis

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Title Polyclonal Antibodies to the Coat Protein of Carnation etched ring virus
Expressed in Bacterial System: Production and Use in Immunodiagnosis
 
Creator Hallan, Vipin
Raikhy, G
Kulshrestha, S
Zaidi, A A
 
Subject Plant viruses
 
Description Carnation etched ring virus (CERV), is the most
widespread virus in carnation cultivars after Carnation
mottle virus. It’s incidences has been reported worldwide.
It has double stranded DNA genome with the
length of �8 kbp. Primers were designed for CERV
coat protein gene (1482 bp) amplification and directional
and inframe cloning in expression vector, pET-
28a(+) (Novagen, USA), using Escherichia coli strain
BL 21 strain competent cells. Expression conditions
for maximum recovery of soluble recombinant protein
was standardized. The in vitro expressed protein was
purified and was used as an antigen for raising antisera.
Both intramuscular and sub-cutaneous routes
were used separately for antisera production and the
antisera was purified. Some of the antisera was used
for enzyme conjugate preparation. This antiserum and
conjugate were then used for formulation of an
ELISA-based diagnostic kit for CERV detection. Its
properties were compared with the commercially available
kit. In all cases, with both glasshouse and field
material, the antibodies had good detectability and
specificity. These antibodies combine specificity to the
target protein and versatility with regard to all the
more important serological techniques.
 
Date 2007
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://ihbt.csircentral.net/218/1/jph.pdf
Hallan, Vipin and Raikhy, G and Kulshrestha, S and Zaidi, A A (2007) Polyclonal Antibodies to the Coat Protein of Carnation etched ring virus Expressed in Bacterial System: Production and Use in Immunodiagnosis. Journal of Phytopathology, 155. pp. 616-622.
 
Relation http://ihbt.csircentral.net/218/