Preparation of Antisera Against Four Proteins Encoded by Cotton Leaf Curl Multan Virus

LIN Dan; LIN Wenzhong; SHANG Pengxiang; LIN Wenwu; DING Xinlun; WU Zujian; ZHANG Jie

doi:10.19303/j.issn.1008-0384.2021.11.014

Volume 36 Issue 11

Nov. 2021

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Article Navigation > Fujian Journal of Agricultural Sciences > 2021 > 36(11): 1359-1364

LIN D, LIN W Z, SHANG P X, et al. Preparation of Antisera Against Four Proteins Encoded by Cotton Leaf Curl Multan Virus [J]. Fujian Journal of Agricultural Sciences，2021，36（11）：1359−1364 doi: 10.19303/j.issn.1008-0384.2021.11.014

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LIN D, LIN W Z, SHANG P X, et al. Preparation of Antisera Against Four Proteins Encoded by Cotton Leaf Curl Multan Virus [J]. Fujian Journal of Agricultural Sciences，2021，36（11）：1359−1364 doi: 10.19303/j.issn.1008-0384.2021.11.014

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Preparation of Antisera Against Four Proteins Encoded by Cotton Leaf Curl Multan Virus

doi: 10.19303/j.issn.1008-0384.2021.11.014

Institute of Plant Virology, Fujian Agriculture and Forestry University/State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops, Fuzhou, Fujian　350002, China

Received Date: 2021-08-16
Rev Recd Date: 2021-09-28

Available Online: 2021-12-30

Publish Date: 2021-11-28

Abstract

Abstract

Objective Antisera of V1, V2, C1, and C4 proteins encoded by the invasive cotton leaf curl Multan virus (CLCuMuV) were prepared for quarantine at entry ports and study on the pathogenic mechanism of the virus. Method Using a prokaryotic expression technology and IPTG induction, V1, V2, C1, and C4 were expressed and the specificity analyzed by western blot. Separately, New Zealand white rabbits were immunized with V1, V2, and C1, while mice with C4 to prepare the antisera. Specificity of the antisera was subsequently verified by western blot. Result The target gene fragments of V1 (768 bp), V2 (363 bp), C1 (1 089 bp), and C4 (300 bp) were amplified by PCR and connected to the respective prokaryotic expression vectors. The SDS-PAGE and western blot showed that the 4 proteins were successfully expressed at 34, 16, 44, and 13 kDa locations with high specificity on the respective antiserum. Conclusion The antisera of CLCuMuV V1, V2, C1, and C4 were successfully prepared for quarantine inspection and study on the pathogenic mechanism of the invasive virus.
- cotton,
- cotton leaf curl Multan virus,
- prokaryotic expression,
- western blot,
- antiserum

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References(26)

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