畜牧与饲料科学2026，Vol.47Issue(3)：23-29,7.DOI:10.12160/j.issn.1672-5190.2026.03.003
bta-miR-19b靶向BCL3的验证及其在牛初乳免疫调控中的功能分析
Validation of bta-miR-19b Targeting BCL3 and Its Functional Analysis in Bovine Colostrum Immune Regulation
摘要
Abstract
[Objective]To validate the targeting relationship between bta-miR-19b and the B-cell CLL/lymphoma 3(BCL3)gene and to elucidate the molecular function and regulatory mechanism by which bta-miR-19b mediates immune regulation in bovine colostrum.[Methods]The intersection of TargetScan and miRanda predictions was used to screen the immune-related target genes of bta-miR-19b,and the minimum free energy(MFE)of binding between bta-miR-19b and immune-related candidate genes IL7R,EGFR,TNF,BCL3,NME7,MAPK8 and MAP3K1 was predicted using the RNAhybrid website.The wild-type(WT)and mutant(MT)fragments were synthesized using the predicted binding site of the 3'UTR of the BCL3 gene as template to construct pmirGLO luciferase reporter vectors.The bta-miR-19b mimics(chemically synthesized double-stranded RNA molecules that mimic endogenous miRNA functions)and NC control(NC-mimics)were synthesized and co-transfected with the reporter vector into 293T cells for target validation.The biological processes and key pathways of bta-miR-19b-BCL3 axis in the immune regulation of bovine colostrum were screened by bioinformatics methods.[Results]The binding free energy of bta-miR-19b with BCL3 was the lowest,suggesting that there was a potential binding of BCL3.The results of dual luciferase reporter gene assay confirmed that bta-miR-19b could specifically bind to the 3'UTR region of the BCL3 gene.The bta-miR-19b-BCL3 regulatory axis was enriched in 42 GO terms,of which 24 were immune-related biological processes.The KEGG pathways involving the bta-miR-19b-BCL3 regulatory axis included tumor necrosis factor TNF signaling pathway and C-type lectin receptor signaling pathway.[Conclusion]bta-miR-19b can target the 3'UTR of BCL3.Given that BCL3 is a regulatory factor of the NF-κB signaling pathway,it is speculated that the bta-miR-19b-BCL3-NF-κB-TNF regulatory axis may be one of the core molecular mechanisms for bovine colostrum to exert immune function,and the specific mechanism needs to be validated by functional experiments.关键词
bta-miR-19b/BCL3/双荧光素酶报告基因实验/牛初乳/免疫调控Key words
bta-miR-19b/BCL3/double luciferase reporter gene assay/bovine colostrum/immune regulation分类
农业科技引用本文复制引用
孔繁霞,王高峰,赵濛,吴美灵,王乐乐..bta-miR-19b靶向BCL3的验证及其在牛初乳免疫调控中的功能分析[J].畜牧与饲料科学,2026,47(3):23-29,7.基金项目
内蒙古自然科学基金项目(2025LHMS03065) （2025LHMS03065）
