现代畜牧兽医Issue(3)：26-31,6.DOI:10.20154/j.cnki.issn1672-9692.2025.03.006
发热伴血小板减少综合征病毒核蛋白的原核表达及多克隆抗体制备
Protein expression and antibody preparation of the nucleoprotein of sever fever with thrombocytopenia syndrome virus
摘要
Abstract
This study aimed to express recombinant severe fever with thrombocytopenia syndrome virus(SFTSV)nucleoprotein(NP)and immunize mice to prepare polyclonal antibodies,laying the foundation for the development of SFTSV diagnostic reagents.The NP gene of SFTSV was amplified and cloned into the pET-28a prokaryotic expression plasmid.Protein expression and purification were performed under optimized conditions.Mice were immunized to generate specific antibodies,and the antibody titer was evaluated by ELISA.The specificity of the antibodies was further analyzed using Western blot and immunofluorescence assay(IFA).Results showed that the 738 bp SFTSV-NP gene was successfully amplified and ligated into the pET-28a plasmid,as confirmed by colony PCR and sequencing.By optimizing expression conditions,maximum protein yield(27 kDa)was achieved at 28℃with 0.1 mmol/L IPTG induction.Large-scale expression and purification were subsequently performed.ELISA confirmed that the antibody titer met experimental requirements,while Western blot and IFA demonstrated excellent specificity.This study successfully purified SFTSV-NP protein and prepared polyclonal antibodies,providing a basis for further research on SFTSV surveillance and functional studies.关键词
发热伴血小板减少综合征病毒(SFTSV)/核蛋白/原核表达/多克隆抗体Key words
Severe fever with thrombocytopenia syndrome virus(SFTSV)/Nucleoprotein/Prokaryotic expression/Polyclonal antibodies分类
农业科技引用本文复制引用
陆桦葆,程子文,胡瀛方,王俊竹,张晓茹,孙荟迪,王大为,杨宝玲..发热伴血小板减少综合征病毒核蛋白的原核表达及多克隆抗体制备[J].现代畜牧兽医,2025,(3):26-31,6.基金项目
锦州医科大学大学生创新创业训练计划项目(X202210160050、X202210160022) （X202210160050、X202210160022）
辽宁省教育厅基本科研项目(JYTQN2023433、JYTQN2023435) （JYTQN2023433、JYTQN2023435）
锦州医科大学-博泽青年科技人才托举项目(JYQT202303、JYBZQT2109) （JYQT202303、JYBZQT2109）
辽宁省自然科学基金博士科研启动基金计划项目(2022-BS-325、2022-BS-323) （2022-BS-325、2022-BS-323）
