中国兽医科学2025，Vol.55Issue(1)：33-38,6.DOI:10.16656/j.issn.1673-4696.2024.0218
猪圆环病毒4型Cap蛋白多克隆抗体的制备
Preparation of polyclonal antibody of porcine circovirus type 4 Cap protein
摘要
Abstract
Using the synthesized full-length genomic DNA of PCV4 as a template,the genes PCV4 Cap and dCap were amplified and cloned into the prokaryotic expression vector pET-28a to obtain recombinant plasmids pET-28a-PCV4-Cap and pET-28a-PCV4-dCap.After digestion and sequencing,the recombinant plasmids were transformed into E.coli BL21 competent cells.After induction by IPTG,the fusion protein His-PCV4-dCap of nearly 28 kDa was successfully expressed and purified by Ni-NTA resin affinity chro-matography.BALB/c mice were immunized three times by subcutaneous multi-point injection to prepare spe-cific polyclonal antibody against PCV4 Cap protein.Western-blotting and indirect immunofluorescence(IFA)results showed that the polyclonal antibody exhibited high reactivity with prokaryotic and eu-karyotic expression of PCV4 Cap.The polyclonal antibody against PCV4 Cap protein prepared in this study lays a foundation for establishing a rapid epidemiological investigation of PCV4 infection,and studying the function of PCV4 Cap and revealing the mechanism of PCV4 pathogenicity.关键词
猪圆环病毒4型/衣壳蛋白/原核表达/多克隆抗体Key words
porcine circovirus type 4/capsid protein/prokaryotic expression/polyclonal antibody分类
畜牧业引用本文复制引用
戴贝凝,周贝怡,周琳怡,蓝小媛,王如嘉,张倩倩,周建卫..猪圆环病毒4型Cap蛋白多克隆抗体的制备[J].中国兽医科学,2025,55(1):33-38,6.基金项目
"十四五"重点研发项目课题(2022YFD1800303) （2022YFD1800303）
江苏省基础研究计划(自然科学基金)(BK20210807) （自然科学基金）
江苏高校优势学科建设工程资助项目(PAPD) （PAPD）
扬州大学大学生科创基金项目(X20220681_2022) （X20220681_2022）
