Target Site Prediction and Validation of BLV-miR-B1-5p Targeting Bovine MUC1
Bovine leukemia virus(BLV)can persistently infect the mammary epithelial cells of dairy cows downstream through the blood and lymphatic systems,which can lead to a decrease in cellular antimicrobial capacity.BLV-encoded miRNAs(BLV-miRNAs)are functional components necessary for BLV replication and pathogenesis.In order to explore whether BLV-miR-B1-5p encoded by BLV participates in inhibiting the antibacterial defense barrier of breast epithelial cells by targeting bovine mucin 1(MUC1)gene,in this study,target sites were first predicted by the software of StarMir,and then verified by cell transfection,dual luciferase report experiments,qRT-PCR and Western Blot.The predicted target sites showed that BLV-miR-B1-5p had a complete binding site with MUC1 3′UTR,with a degree of freedom of-35.6 kcal·mol-1.The results of dual luciferase report experiment showed that BLV-miR-B1-5p could significantly reduce the fluorescence value of MUC1-3UTR-WT(P＜0.001);And the results of miRNA cell transfection,qRT-PCR,and Western Blot in vitro validation showed that BLV-miR-B1-5p can inhibit the expression of MUC1 mRNA and protein in bovine mammary epithelial cells.Based on the above results,BLV-miR-B1-5p can target and inhibit bovine MUC1,and this conclusion provides data support for BLV promote pathogen adhesion on breast epithelial cells.
bovine leukemia virusMUC1mammary epithelial cells of dairy cowsmiRNA
万方数据
维普
