中国动物传染病学报2026，Vol.34Issue(3)：149-154,6.
非洲猪瘟病毒B318L蛋白的多克隆抗体制备及功能初探
Preparation of Polyclonal Antibodies to ASFV B318L Protein and Preliminary Function Analysis
摘要
Abstract
In this study,the ASFV genome was used as a template for high-fidelity enzyme-mediated PCR amplification followed by enzymatic cleavage and ligation to construct a prokaryotic expression plasmid pET-30a-B318L-C.After sequencing validation,the plasmid was transformed into competent BL21(DE3)cells,resulting in successful expression of a 18 kDa ASFV B318L truncated protein predominantly present in inclusion bodies after induction with IPTG.The target protein was purified to high purity via magnetic bead-based affinity chromatography.The purified protein was subsequently used to immunize mice to obtain polyclonal antibodies.Results of IFA and WB demonstrated that polyclonal antibodies present in mouse sera were able to recognize the expression of B318L protein in ASFV-infected cells,indicating satisfactory antigenicity and specificity.Furthermore,eukaryotic transfection and click chemistry assays revealed that ASFV pB318L increased the prenylation level of host protein RhoA.This work provided valuable biological materials and theory for elucidating the functions of African swine fever virus proteins.关键词
非洲猪瘟病毒/B318L/原核表达/多克隆抗体Key words
African swine fever virus/B318L/prokaryotic expression/polyclonal antibody分类
农业科技引用本文复制引用
周州,曾凡亮,胡松洋,李智晟,谢振华,刘英楠,陈鸿军..非洲猪瘟病毒B318L蛋白的多克隆抗体制备及功能初探[J].中国动物传染病学报,2026,34(3):149-154,6.基金项目
中国农业科学院青年创新专项(Y2024QC26) （Y2024QC26）
国家自然科学基金面上项目(32170161) （32170161）
