Preparation and Application of Polyclonal Antibodies Recognizing CP Protein in Pepper Mottle Virus

WANG Xin; YE Qian; LYU Xiaoyuan; TIAN Peijie; ZHANG Songbai; ZHANG Yu; ZHANG Deyong; LUO Xiangwen

doi:10.19303/j.issn.1008-0384.2022.012.011

Volume 37 Issue 12

Dec. 2022

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Article Navigation > Fujian Journal of Agricultural Sciences > 2022 > 37(12): 1595-1600

WANG X, YE Q, LYU X Y, et al. Preparation and Application of Polyclonal Antibodies Recognizing CP Protein in Pepper Mottle Virus [J]. Fujian Journal of Agricultural Sciences，2022，37（12）：1595−1600 doi: 10.19303/j.issn.1008-0384.2022.012.011

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WANG X, YE Q, LYU X Y, et al. Preparation and Application of Polyclonal Antibodies Recognizing CP Protein in Pepper Mottle Virus [J]. Fujian Journal of Agricultural Sciences，2022，37（12）：1595−1600 doi: 10.19303/j.issn.1008-0384.2022.012.011

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Preparation and Application of Polyclonal Antibodies Recognizing CP Protein in Pepper Mottle Virus

doi: 10.19303/j.issn.1008-0384.2022.012.011

WANG Xin^1
,,
YE Qian²,
LYU Xiaoyuan³,
TIAN Peijie²,
ZHANG Songbai^{1, 2},
ZHANG Yu²,
ZHANG Deyong^{1, 2},
LUO Xiangwen^{1, 2
,
,}

1.
Longping Branch, Biology College, Hunan University, Changsha, Hunan　410125, China
2.
Plant Protection Institute, Hunan Academy of Agricultural Sciences, Changsha, Hunan　410125, China
3.
Technical Center of Changsha Customs, Changsha, Hunan　410300, China

Received Date: 2022-09-01
Rev Recd Date: 2022-09-16

Available Online: 2022-12-28

Publish Date: 2022-03-28

Abstract

Abstract

Objective A specific, rapid method based on the polyclonal antibody prepared using purified recombinant CP protein for detecting a typical Potyvirus, pepper mottle virus (PepMoV), on Capsicum annuum L. was developed to assess the distribution, occurrence ratio, and pathogenicity of the disease in China. Methods The 822 bp of CP was amplified by specific RT-PCR using the total RNA of PepMoV-infected chili peppers. It was cloned into prokaryotic expressing plasmid pET28α and expressed in E. coli DH5α. The recombinant CP protein was purified by Ni-NTA chromatography and used as the antigen to prepare the polyclonal antibodies to be verified by ID-ELISA and western blotting. Results The purified recombinant CP protein was approximately 37 kDa, and the prepared polyclonal antibody verified to specifically recognize PepMoV CP protein. The ID-ELISA method detected 20.00% PepMoV infection on field chili pepper specimens in Hunan and 43.33% in Guizhou. Conclusion The established specific and rapid detection method based on the polyclonal antibody against CP protein of PepMoV was applied to survey the disease spreading on chili peppers in China. It provided a tool for further studies on the CP protein.
- Pepper mottle virus,
- CP gene,
- polyclonal antibody,
- ID-ELISA,
- Rapid detection

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References(22)

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