中国动物传染病学报2026，Vol.34Issue(2)：115-124,10.
IL15多克隆抗体的制备及其在猪繁殖与呼吸综合征病毒活载体中的表达
Preparation of Polyclonal Antibodies Against Interleukin-15 Protein and Its Expression in Porcine Reproductive and Respiratory Syndrome Virus Live Vector
摘要
Abstract
This study aimed to prepare polyclonal antibodies against IL15 and validate their use with the plasmid pCDNA3.1-IL15 and the recombinant virus rPRRSV-IL15.Firstly,the plasmid pET-IL15 was constructed for prokaryotic expression with IPTG induction.The expressed recombinant protein was confirmed in SDS-PAGE to determine its reactivity and antigenicity.The purified recombinant IL15 protein was used to immunize rabbits to generate polyclonal antibodies.Meanwhile,the IL15 gene was inserted into the infectious clone of the highly pathogenic porcine reproductive and respiratory syndrome virus(PRRSV)strain vHuN4-F112 using reverse genetics techniques,resulting in the rescue of the recombinant virus rPRRSV-IL15,which stably expresses the IL15 protein.Subsequently,the plasmid pCDNA3.1-IL15 expressing IL15 protein was transfected into HEK 293T cells,and rPRRSV-IL15 infected MARC-145 cells and PAMs.The prepared antibodies were used to validate the IL15 expression in the transfected cells and rPRRSV-IL15-infected cells.The results demonstrated that the prepared polyclonal antibodies against IL15 showed good reactivity and specificity,capable of recognizing and binding to IL15 protein in the cells transfected with pCDNA3.1-IL15 and infected with rPRRSV-IL15.This study laid an important foundation for further investigating the biological functions of IL15 protein and its role in immune regulation.The recombinant virus rPRRSV-IL15 also represented an attempt to develop a novel immunoenhancing PRRSV attenuated vaccine.关键词
猪繁殖与呼吸综合征病毒/病毒活载体/IL15蛋白/多克隆抗体Key words
Porcine reproductive and respiratory syndrome virus/live viral vector/IL15 protein/polyclonal antibody分类
农业科技引用本文复制引用
郭子强,童光志,高飞,李佳乐,罗梦,唐银保,帅文娜,李丽薇,周艳君,童武,姜一峰..IL15多克隆抗体的制备及其在猪繁殖与呼吸综合征病毒活载体中的表达[J].中国动物传染病学报,2026,34(2):115-124,10.基金项目
上海市自然基金(21ZR1476900) （21ZR1476900）
国家重点研发计划(2022YFD1800804,2021YFD1801401) （2022YFD1800804,2021YFD1801401）
国家自然科学基金(32373050) （32373050）
中国农业科学院科技创新工程(CAAS-CSLPDCP-202402) （CAAS-CSLPDCP-202402）
上海市兽医生物技术重点实验室开放基金(BD1500010) （BD1500010）
