中国兽医科学2025，Vol.55Issue(3)：314-320,7.DOI:10.16656/j.issn.1673-4696.2025.0040
猪细小病毒NS1基因编码蛋白的原核表达及多克隆抗体的制备
Prokaryotic expression of protein encoded by porcine parvovirus NS1 gene and preparation of polyclonal antibody
摘要
Abstract
The aim of this study was to prepare a polyclonal antibody to PPV-NS1 and to perform double validation with porcine parvovirus(PPV)and eukaryotic plasmid pcDNA3.1(+)-NS1-3 ×Flag.Firstly,the non-structural protein encoding gene NS1 of PPV was cloned into the prokaryotic expression vector pETDuet,and the prokaryotic plasmid pET-PPV-NS1 was constructed.Then,the prokaryotic plasmid pET-PPV-NS1 was transformed into competent E.coli BL21 cells,induced by 1 mmol/L IPTG,and the NS1 expression was detected by SDS-PAGE and Western-blot,and the results showed that the NS1 gene was well expressed in competent cells.Secondly,the purified NS1 protein was injected into rabbits using a multi-point injection method,and polyclonal antibodies against the recombinant NS1 protein were prepared.Then,the eukaryotic plasmid pcDNA3.1(+)-NS1-3XFlag was transfected into HEK 293T cells.At the same time,protein samples were collected after PPV infected ST cells,and the NS1 polyclonal antibody prepared above was used for validation.The results showed that the prepared NS1 polyclonal antibody had good reactivity and specificity.关键词
猪细小病毒/NS1蛋白/多克隆抗体/原核表达Key words
porcine parvovirus/NS1 protein/polyclonal antibodies/prokaryotic expression分类
农业科技引用本文复制引用
帅文娜,李兆龙,高飞,郭子强,李佳乐,罗梦,李丽薇,周艳君,姜一峰,童武,童光志..猪细小病毒NS1基因编码蛋白的原核表达及多克隆抗体的制备[J].中国兽医科学,2025,55(3):314-320,7.基金项目
国家重点研发计划项目(2022YFD1800800,2021YFD1801401) （2022YFD1800800,2021YFD1801401）
上海市自然科学基金项目(21ZR1476900) （21ZR1476900）
国家自然科学基金项目(32373050,32072861) （32373050,32072861）
中国农业科学院科技创新工程项目(CAAS-CSLPDCP-202402) （CAAS-CSLPDCP-202402）
上海市兽医生物技术重点实验室开放基金项目(BD1500010) （BD1500010）
