Preparation and Application of Polyclonal Antibody of Chilli Veinal Mottle Virus Capsid

JIANG Jun; LI Xiquan; DAI Fei; ZHANG Fuqiang; LU Zhou; WANG Hancheng; SHI Caihua; FANG Shouguo; ZHANG Songbai; PAN Shouhui

doi:10.19303/j.issn.1008-0384.2024.02.009

Volume 39 Issue 2

Feb. 2024

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Article Navigation > Fujian Journal of Agricultural Sciences > 2024 > 39(2): 193-198

JIANG J, LI X Q, DAI F, et al. Preparation and Application of Polyclonal Antibody of Chilli Veinal Mottle Virus Capsid [J]. Fujian Journal of Agricultural Sciences，2024，39（2）：193−198 doi: 10.19303/j.issn.1008-0384.2024.02.009

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JIANG J, LI X Q, DAI F, et al. Preparation and Application of Polyclonal Antibody of Chilli Veinal Mottle Virus Capsid [J]. Fujian Journal of Agricultural Sciences，2024，39（2）：193−198 doi: 10.19303/j.issn.1008-0384.2024.02.009

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Preparation and Application of Polyclonal Antibody of Chilli Veinal Mottle Virus Capsid

doi: 10.19303/j.issn.1008-0384.2024.02.009

1.
MARA Key Laboratory of Sustainable Crop Production in the Middle Reaches of the Yangtze River (Co-construction by Ministry and Province)/Yangtze University, Jingzhou, Hubei　434025, China
2.
Anshun Branch, Guizhou Tobacco Company, Anshun, Guizhou　561000, China
3.
Guizhou Academy of Tobacco Science, Guiyang, Guizhou　550081, China

Received Date: 2023-04-14
Rev Recd Date: 2023-11-09

Available Online: 2024-03-28

Publish Date: 2024-02-28

Abstract

Abstract

Objective A highly specific polyclonal antibody of the capsid of chilli veinal mottle virus (ChiVMV) was prepared for field detection and quarantine inspection on infected Solanaceae plants. Methods The full-length (861 bp) and partial fragment (396 bp) of the gene from a ChiVMV GZ-Tabacco isolate were amplified using RT-PCR, recombined into the prokaryotic expression vector pET28a, and transformed into E. coil BL21 for induced expression. After chromatographic isolation and dialysis purification, the products were used to immunize Dahl rabbits for the antibody preparation. Potency and specificity of the candidate antibodies were evaluated by ELISA and western blot. Results Two polyclonal antibodies, namely antiCP^1-287aa and antiCP^1-132aa with the titers of 1∶6400 and 1∶12800, respectively, were obtained. Both positively detected the antigen as shown by western blot. But the indirect ELISA on the field strains revealed that antiCP^1-132aa specifically detected ChiVMV not potato virus Y (PVY), whereas antiCP^1-287aa failed to differentiate the two. Conclusion The identified polyclonal antibody, antiCP^1-132aa was highly specific in detecting ChiVMV in field tests. It could become a useful tool for further studies on the infectious virus of chilli peppers.
- Chilli veinal mottle virus,
- capsid,
- prokaryotic expression,
- chromatographic purification,
- polyclonal antibody

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References(24)

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