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Expression of recombinant Chrysanthemum virus B coat protein for raising polyclonal antisera

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Title Expression of recombinant Chrysanthemum virus B coat
protein for raising polyclonal antisera
 
Creator Singh, L
Hallan, Vipin
Raja, Ram
Zaidi, A A
 
Subject Plant sciences
Plant viruses
 
Description Chrysanthemum virus B (CVB genus Carlavirus,
family Flexiviridae) is one of the major viral pathogens of
chrysanthemum. This virus occurs worldwide, is a potential
threat to the floriculture industry and hence is a quarantine
pathogen. CVB has a positive sense single-stranded RNA
(ssRNA) genome ~8.8 kb. The coat protein gene of CVB was
amplified by RT-PCR, cloned and over expressed in E. coli
BL21. The protein (CP) was expressed as a fusion protein
with Glutathione S-Transferase (GST). Fused protein with
GST was purified by GST tagged affinity chromatography
and polyclonal (but monospecific) antibodies from rabbits
immunized with the fusion protein was used for formulation
of ELISA based diagnostic kit for CVB detection. The
antisera produced showed specific reaction to CVB from
infected chrysanthemums, Nicotiana glutinosa, Nicotiana
clevelandii and Chenopodium quinoa at dilution of 1:1000
in ELISA. Results obtained were comparable (some times
better) than commercial kit. The IgG against CVB performed
favorably in specificity and sensitivity to the virus. Findings
present a procedure for production of antibodies for CVB
indexing of chrysanthemums propagative and mother stock
materials to provide the disease free planting material.
 
Date 2011
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://ihbt.csircentral.net/623/1/572_2011_9-12-2011.pdf
Singh, L and Hallan, Vipin and Raja, Ram and Zaidi, A A (2011) Expression of recombinant Chrysanthemum virus B coat protein for raising polyclonal antisera. Journal of Plant Biochemistry and Biotechnology, 20. pp. 96-101.
 
Relation http://ihbt.csircentral.net/623/