动物医学进展2025，Vol.46Issue(10)：49-55,7.
山羊痘病毒融合蛋白P32-L1R的原核表达及多克隆抗体制备
Prokaryotic Expression of Fusion Protein P32-L1R in Goatpox Virus and Preparation of Polyclonal Antibody
摘要
Abstract
The aim was to perform bioinformatics analysis and prokaryotic expression of the truncated fu-sion structural protein P32-L1R of the Goatpox virus(GTPV)and to prepare polyclonal antibodies.Analy-sis of the P32-L1R gene using multiple bioinformatics tools,identified by SDS-PAGE and Western blot.A Polyclonal antibody was prepared by immunizing BALB/c mice with purified P32-L1R protein.The serum titer and immunoreactivity of the polyclonal antibody were detected by indirect ELISA and Western blot.The results showed that P32-L1R encoded 458 amino acids,and the protein size was about 51 kDa.P32-L1R is a basic protein,and its molecular formula is C2283H3614N594O707S17,which belongs to hydrophilic pro-tein with good stability.P32-L1R has no signal peptide region or transmembrane domain,with 16 antigenic determinants,57 phosphorylation sites,and 3 N-glycosylation sites.The secondary structure of the P32-L1R protein contained α-helix,β-turn,extended chain,and random coil.pCold I-P32-L1R was transformed into E.coli and induced to express the recombinant protein P32-L1R,which mainly existed in the form of inclusion bodies.Immunization of BALB/c mice produced polyclonal antibodies with a potency of 1∶819200.This study can be used as a reference for establishing GTPV detection methods and for the develop-ment of GTPV subunit vaccines.关键词
山羊痘病毒/P32-L1R基因/原核表达/多克隆抗体Key words
Goatpox virus/P32-L1R gene/Prokaryotic expression/Polyclonal antibody分类
农业科技引用本文复制引用
王永璇,胡茜,胡鹏飞,瞿正文,朱二鹏,王乃秀,文明,肖琨,程振涛..山羊痘病毒融合蛋白P32-L1R的原核表达及多克隆抗体制备[J].动物医学进展,2025,46(10):49-55,7.基金项目
贵州省科技计划项目(黔科合成果[2023]一般077) （黔科合成果[2023]一般077）
贵州省科技计划项目(黔科合平台人才[2021]5646号) （黔科合平台人才[2021]5646号）
