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.

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Abstract

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.

Item Type:	Article
Uncontrolled Keywords:	CVB detection . Indexing . Polyclonal antisera . Expression . Diagnostics
Subjects:	Plant sciences Plant viruses
Divisions:	UNSPECIFIED
Depositing User:	Dr. Aparna Maitra Pati
Date Deposited:	30 Dec 2011 03:04
Last Modified:	30 Dec 2011 03:04
URI:	http://ihbt.csircentral.net/id/eprint/623

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