Cloning,Tissue Expression and Bioinformatics Analysis of miR-222 Precursor Sequence in Hu Sheep
To explore the role of miR-222 in the growth,development and reproductive function in this experiment,the precursor sequence of miR-222 in Hu sheep was cloned through polymerase chain reaction.Real-time fluorescent quan-titative technology was used to detect the expression pattern of miR-222 in the hypothalamus,pituitary,and ovary of Hu sheep.DNAStar was used to conduct a conservative analysis of the mature sequence of miR-222.MEGA 11 was used to establish a gene evolutionary tree and analyze the phylogenetic relationship of miRNA-222 among various species.miR-PathD8,TargetScan,and StarBase software were used to predict the potential target genes of miR-222 in Hu sheep,and GO functional enrichment analysis and KEGG pathway enrichment analysis were conducted on the target genes.The re-sults indicated that the precursor sequence of miR-222 in Hu sheep was successfully obtained.The relative expression level of miR-222 in the ovary of Hu sheep was significantly higher than that in the hypothalamus and pituitary.The ma-ture sequence of miR-222 in Hu sheep was the most closely related to that of macaques,exhibiting a high degree of conservation among different species.Prediction results of the target genes of miR-222 in Hu sheep revealed that some of the potential target genes related to reproduction included PIK3R1,IGF1,GNAI2 and THBS1.The enriched signaling pathways included the PI3K-AKT and Wnt signaling pathways,which both played crucial roles in the regulation of ani-mal reproduction.Therefore,miR-222 may play a crucial regulatory role in the ovarian development of Hu sheep.
Hu sheepmiRNAprecursor sequencebioinformatics analysistarget genes
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