佛山科学技术学院学报（自然科学版）2026，Vol.44Issue(1)：1-7,7.
猪圆环病毒2型去核定位信号Cap蛋白的原核表达及多克隆抗体制备
Prokaryotic expression of Capsid Protein without nuclear localization signal of Porcine circovirus 2 and preparation for its polyclonal antibody
摘要
Abstract
To generate polyclonal antibodies targeting the Cap protein of Porcine Circovirus type 2(PCV2),a Cap gene fragment without the nuclear localization signal(NLS)was amplified using the pMD18-T-ORF2 plasmid as a template.This fragment was subsequently cloned into the prokaryotic expression vector pET-28a(+)to construct the recombinant plasmid pET-28a-N-Cap.The recombinant plasmid was then transformed into Escherichia coli Rosetta(DE3)competent cells.After induction with IPTG,SDS-PAGE analysis revealed that the expressed recombinant protein was predominantly present in an insoluble form.The target protein was purified via nickel-affinity chromatography and used as an antigen to immunize rabbits for the production of rabbit-derived polyclonal antibodies.The resulting antibody titer reached up to 1∶204,800,as determined by serological testing.Furthermore,indirect immunofluorescence assay(IFA)and Western blot analysis confirmed that the obtained polyclonal antibodies specifically recognized PCV2.These findings indicate that the NLS-deleted Cap protein expressed in a prokaryotic system retains strong immunogenicity,and the acquired polyclonal antibodies can be utilized in assays related to the Cap protein.关键词
猪圆环病毒 2型/衣壳蛋白/原核表达/核定位信号/多克隆抗体Key words
Porcine circovirus type 2/Capsid protein/prokaryotic expression/nuclear localization signal/polyclonal antibody分类
农业科技引用本文复制引用
陈丽莎,冯幸瑶,唐子锐,胡凯茵,郭锦玥..猪圆环病毒2型去核定位信号Cap蛋白的原核表达及多克隆抗体制备[J].佛山科学技术学院学报（自然科学版）,2026,44(1):1-7,7.基金项目
广东省基础与应用基础研究基金项目(2022A1515140057) （2022A1515140057）
