海南大学学报（自然科学版）2025，Vol.43Issue(2)：114-120,7.DOI:10.15886/j.cnki.hdxbzkb.2024050601
卵形鲳鲹干扰素调节因子IRF3的原核表达和多克隆抗体的制备
Prokaryotic expression and polyclonal antibody preparation of Trachinotus ovatus interferon regulatory factor IRF3
摘要
Abstract
Interferon regulatory factor 3(IRF3)regulates the expression of interferon(IFN),and which mediates the host defense against pathogen infection.In order to obtain the recombinant protein of Trachinotus ovatus IRF3(TroIRF3)and its polyclonal antibody,in the report,the pET32a(+)/Escherichia coli BL21(DE3)expression system was used for the inducible expression of TroIRF3.Under the conditions,0.1 mmol·L-1 IPTG,24 h,20℃,the soluble recombinant protein rTroIRF3 was obtained.The molecular mass of the rTroIRF3 protein was 51 kDa.The rTroIRF3 protein was purified by nickel affinity chromatography.The purified fusion protein rTroIRF3 was used as an antigen to immunize mice for the preparation of the polyclonal antibody.ELISA and western blotting were performed.The results indicated that the total titer of the polyclonal antibody of rTroIRF3 was 1∶25 600,the polyclonal antibody could bind specifically to the recombinant protein,and which can be used as a good experimental material for the subsequent mechanism analysis of TroIRF3 during pathogen infection.关键词
卵形鲳鲹/IRF3/原核表达/多克隆抗体Key words
Trachinotus ovatus/IRF3/prokaryotic expression/polyclonal antibody分类
农业科技引用本文复制引用
吴莹,周永灿,孙云..卵形鲳鲹干扰素调节因子IRF3的原核表达和多克隆抗体的制备[J].海南大学学报（自然科学版）,2025,43(2):114-120,7.基金项目
国家自然科学基金地区项目(42066007) （42066007）
