中国兽医科学2026，Vol.56Issue(4)：539-545,7.DOI:10.16656/j.issn.1673-4696.2026.0067
猪源核糖体RNA加工蛋白1B的多克隆抗体制备及与核糖体和核仁共定位研究
Preparation of polyclonal antibodies against porcine RRP1B and investigation of its co-localization with ribosomes and nucleoli
摘要
Abstract
This study aimed to obtain polyclonal antibodies against porcine ribosome RNA process-ing protein 1B(RRP1B)and investigate its co-localization with ribosomes and nucleoli.The full-length RRP1B gene was amplified from IPEC-J2 using RT-PCR and cloned into the eukaryotic expression vector pcDNA3.1.The recombinant plasmid pcDNA3.1(+)-RRP1B-His was constructed.After sequencing verifica-tion,the plasmid was transfected into HEK-293T and IPEC-J2.Expression of RRP1B was detected by indirect immunofluorescence assay(IFA)and Western-blot.The recombinant His-RRP1B was purified by nickel affinity chromatography and used as an immunogen to generate polyclonal antibodies in BALB/c mice via subcutaneous immunization.Antibody titers were determined by Western-blot and IFA to evaluate immunogenicity.Meanwhile,the prepared polyclonal antibody was applied to investigate the co-localization of RRP1B with ribosomes and nucleoli.The results showed that the recombinant plasmid pcDNA3.1(+)-RRP1B-His was successfully expressed in both HEK-293T and IPEC-J2.Immunization with the recombinant RRP1B induced high-level antibody production in mice,with titers reaching above 1∶2 000.Immunofluorescence co-localization assays revealed that RRP1B was localized in the nucleoli,with no significant co-loca-lization observed with ribosomes.This study lays a foundation for further investigation on the biological functions of RRP1B.关键词
RRP1B/多克隆抗体/核糖体/核仁/共定位Key words
RRP1B/polyclonal antibody/ribosome/nucleolus/co-localization分类
农业科技引用本文复制引用
薛晓峰,徐双媛,牛自艺,李彬,李基棕,索朗斯珠..猪源核糖体RNA加工蛋白1B的多克隆抗体制备及与核糖体和核仁共定位研究[J].中国兽医科学,2026,56(4):539-545,7.基金项目
国家自然科学基金项目(32473119) （32473119）
江苏省自然科学基金项目(BK20221432) （BK20221432）
