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Journal of Integrative Agriculture  2023, Vol. 22 Issue (6): 1945-1949    DOI: 10.1016/j.jia.2023.04.012
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Identification of two novel linear epitopes on the p30 protein of African swine fever virus

YU Si-hui1*, SHAN Zhao-meng1*, YANG Jing-jing1, LIU Yi-ning1, WU Chang-de1, ZHANG Zhen-jiang2, ZHU Yuan-mao2, MENG Bo1, ZHAN Jia-xing1, WEN Xue-xia1#, ZHANG Ying1#

1 Key Laboratory of Livestock Infectious Diseases of Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110866, P.R.China

2 State Key Laboratory of Veterinary Biotechnology, National High Containment Facilities for Animal Diseases Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, P.R.China

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摘要  

非洲猪瘟(ASF)的持续流行严重威胁全球养猪业的健康发展。该病目前缺乏可应用于临床的疫苗和药物,因此,早期的精准检测对于ASF的有效防控至关重要。p30蛋白在病毒感染早期表达,免疫原性较好且诱导产生抗体水平较高,这些特性使其成为较为理想的非洲猪瘟病毒ASFV)检测靶点之一。本研究以原核表达的p30蛋白为免疫原,制备了两株可稳定分泌针对ASFV p30蛋白的单克隆抗体(mAb细胞株,分别命名为4F27F3利用Western blotting和间接免疫荧光实验鉴定两株单克隆抗体均能识别病毒感染。亚型鉴定结果显示,4F27F3的重链分别为IgG1IgG2b,轻链均为κ链。随后,通过原核表达一系列截短p30蛋白鉴定4F27F3识别的抗原表位。5轮截短和Western blotting实验鉴定,最终确定4F27F3识别的最短抗原表位分别为118SFETL12276EHQAQEEWNMILHV89。在此基础上,进一步分析两个抗原表位在不同毒株p30蛋白序列中的保守性。序列比对结果表明,GenBank数据库中所有p30蛋白序列均包含118SFETL122 ,而另一抗原表位76EHQAQEEWNMILHV89则在非洲以外地区分离的毒株中保守。本研究制备的两株单克隆抗体及鉴定的抗原表位可为检测方法的开发提供材料和基础,同时也可为病毒致病机理和免疫机制等基础研究提供工具。



Received: 06 January 2023   Online: 15 April 2023   Accepted: 15 March 2023
Fund: 

This study was supported by the National Natural Science Foundation of China (31902258), the Scientific Research Staring Foundation for the Doctors from the Science and Technology Department of Liaoning Province, China (2019-BS-204), and the Major Project of Science and Technology of Liaoning Province (2020JH1/10200003).

About author:  #Correspondence WEN Xue-xia, Tel: +86-24-88487156, E-mail: wenxuexia123@syau.edu.cn; ZHANG Ying, Tel: +86-24-88487156, E-mail: yingzhang18@syau.edu.cn * These authors contributed equally to this study.

Cite this article: 

YU Si-hui, SHAN Zhao-meng, YANG Jing-jing, LIU Yi-ning, WU Chang-de, ZHANG Zhen-jiang, ZHU Yuan-mao, MENG Bo, ZHAN Jia-xing, WEN Xue-xia, ZHANG Ying. 2023. Identification of two novel linear epitopes on the p30 protein of African swine fever virus. Journal of Integrative Agriculture, 22(6): 1945-1949.

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