Preparation and Application of Polyclonal Antibodies Recognizing the Capsid Protein of Chilli Veinal Mottle Virus

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

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JIANG J, LI X Q, DAI F, et al. Preparation and Application of Polyclonal Antibodies Recognizing the Capsid Protein of Chilli Veinal Mottle Virus [J]. Fujian Journal of Agricultural Sciences，2024，39（2）：1−6

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JIANG J, LI X Q, DAI F, et al. Preparation and Application of Polyclonal Antibodies Recognizing the Capsid Protein of Chilli Veinal Mottle Virus [J]. Fujian Journal of Agricultural Sciences，2024，39（2）：1−6

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Preparation and Application of Polyclonal Antibodies Recognizing the Capsid Protein of Chilli Veinal Mottle Virus

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 of 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

Abstract

Abstract

Aim Chilli veinal mottle virus (ChiVMV) is one of the most harmful viruses in the production of solanaceae palnts and a subject of quarantine inspections at ports. The aim of this study is to develop polyclonal antibodies against ChiVMV coat proteins with a high level of specificity, which can then be utilized for the field detection of infected plants. Methods The full-length (861 bp) and partial fragment (396 bp) of the coat protein gene from the ChiVMV GZ-tabacco isolate were amplified using RT-PCR, recombined into the prokaryotic expression vector pET28a, and subsequently transformed into E.coil BL21 for induced expression. After chromatographic isolation and dialysis purification of the expression products, they were used to immunize Dahl rabbits for the preparation of polyclonal antibodies. The antibody potency and specificity were assessed through enzyme-linked immunosorbent assay (ELISA) and Western blot.. Results Two polyclonal antibodies against coat proteins, namely antiCP^1-287aa and antiCP^1-132aa, were successfully prepared with antibody titer of 1:6400 and 1:12800, respectively. Both antibodies, antiCP^1-287aa and antiCP^1-132aa, effectively detected the antigen by Western blot. In indirect ELISA tests of field strains, it was observed that the ChiVMV strain was specifically detectable by antiCP^1-132aa, whereas the Potato virus Y （PVY） strain was not. However, antiCP^1-287aa could not differentiate between these two strains. Conclusion The polyclonal antibody, antiCP^1-132aa , exhibits high specificity, making it suitable for detecting ChiVMV-infected strains in the field. Additionally, it lays the foundation for the functional study of the coat protein of ChiVMV.
- Chilli veinal mottle virus,
- Coat protein,
- Prokaryotic expression,
- Chromatographic purification,
- Polyclonal antibody

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References(24)

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