中国兽医科学2025，Vol.55Issue(7)：913-918,6.DOI:10.16656/j.issn.1673-4696.2025.0129
非洲猪瘟病毒pH108R蛋白的原核表达及其多克隆抗体的制备
Prokaryotic expression of African swine fever virus pH108R protein and preparation of its polyclonal antibodies
摘要
Abstract
The aim of this study was to prepare polyclonal antibodies against the H108R protein(pH108R)of African swine fever virus(ASFV).Initially,the prokaryotic expression plasmid pE-22-H108RΔTR for the recombinant pH108R protein was constructed by deleting the transmembrane region(TR)of pH108R.The successful construction of this plasmid was confirmed through enzymatic analysis.Sub-sequently,the plasmid was transformed into E.coli BL21(DE3)competent cells,and pH108R was expressed fo-llowing IPTG induction.SDS-PAGE analysis revealed that the protein was expressed as inclusion bodies,which was purified using nickel column affinity chromatography and used to immunize guinea pigs for the preparation of polyclonal antibodies.Results from immunoblotting,indirect immunofluorescence,and co-immunoprecipitation assays demonstrated that the prepared polyclonal antibodies exhibited good specificity and could recognize pH108R expressed in ASFV-infected porcine alveolar macrophages.This study provides an effective detection tool for the in-depth investigation of the biological function of ASFV pH108R.关键词
非洲猪瘟病毒/原核表达/pH108R蛋白/多克隆抗体Key words
African swine fever virus/prokaryotic expression/pH108Rprotein/polyclonal antibody分类
农业科技引用本文复制引用
王梦旖,任静静,武正前,王志艺,吴俊煌,甘露露,李丹,尚佑军,郑海学..非洲猪瘟病毒pH108R蛋白的原核表达及其多克隆抗体的制备[J].中国兽医科学,2025,55(7):913-918,6.基金项目
"十四五"国家重点研发计划项目(2021YFD1800100) （2021YFD1800100）
甘肃省重大专项(21ZD3NA001-5,22ZD6NA001) （21ZD3NA001-5,22ZD6NA001）
甘肃省重大人才项目(212068657016) （212068657016）
甘肃省陇原青年英才项目(20230102) （20230102）
