绵羊miR-150靶基因预测及生物信息学分析

doi:10.16431/j.cnki.1671-7236.2021.07.007

摘要/Abstract

摘要： 为探究miR-150在绵羊卵巢中的调控机制，本研究利用生物信息学方法对miR-150进行靶基因预测与分析。通过miRBase数据库获取miR-150成熟序列，进行序列比对和保守性分析；利用Ensemble数据库搜索miR-150前体序列，并利用PROMO在线软件对其上游2 000 bp的启动子区进行转录因子结合位点预测；通过TargetScan和miRwalk在线软件预测并取靶基因的交集，采用DAVID和KOBAS在线软件对预测靶基因进行GO功能和KEGG通路富集分析；利用Networkanalyst在线软件对靶基因进行功能性蛋白的互作分析，绘制miR-150靶基因参与的调控网络；利用YM500V2在线软件分析miR-150靶基因在不同组织和疾病中的差异表达水平。结果显示，miR-150成熟序列在不同物种间具有高度保守性；miR-150启动子区存在p53、ID1、FOXO4等转录因子结合位点。GO功能分析结果显示，主要富集在RNA聚合酶Ⅱ启动子转录的正调控等生物过程，锌离子结合、DNA结合、转录DNA模板等分子功能，以及核、核质、胞质、胞质核周区域等细胞组分。KEGG通路富集结果显示，主要富集在癌症通路、癌症中蛋白聚糖、人乳头瘤病毒感染、乳腺癌、调节干细胞多能性等信号通路。miR-150靶基因互作分析发现，靶基因PIK3R1、PIK3CB、CTNNB1与其他蛋白存在较多靶向关系，参与了雌激素信号通路、细胞衰老、磷脂酰肌醇信号系统、TGF-β等通路。根据YM500V2在线软件分析miR-150在不同组织和疾病中的表达发现，在血液中表达水平最高，在卵巢、胰腺、淋巴和皮肤中表达水平相对较高，在脑和肾上腺皮质中表达水平很低或不表达。通过对绵羊miR-150靶基因的生物信息学分析，为其功能和调控机制研究提供参考依据，为绵羊miR-150在卵巢发育中的调控机制奠定基础。

关键词: 绵羊; miR-150; 靶基因; 生物信息学

Abstract: In order to explore the regulatory mechanism of miR-150 in ovaries of sheep,the target gene prediction and analysis was performed with bioinformatic method.Mature miR-150 sequences were obtained from miRBase database for sequence alignment and conservative analysis.The Ensemble database was used to search the miR-150 precursor sequence,and the online software PROMO was used to predict the transcription factor binding site of miR-150.The target genes of intersection were predicted by TargetScan and miRwalk,and the Go function and KEGG pathway enrichment analysis of the predicted target genes were performed by DAVID and KOBAS online software.The functional protein interactions was analyzed with Networkanayst online tools.YM500V2 software was used to analyze the differential expression levels of miR-150 target genes in different tissues and diseases.The results showed that the miR-150 mature sequences between species were highly conservative.There were many transcription factor binding sites such as p53,ID1 and FOXO4 in the promoter region of miR-150.GO function enrichment analysis results showed that the predicted target genes of miR-150 mainly enriched in the positive regulation of RNA polymerase Ⅱ promoter transcription for biological process,zinc ions binding,DNA binding and transcription of DNA template for molecular functions,nuclear,nuclear and cytoplasmic and perinuclear area of cytoplasm for cellular components.KEGG pathway enrichment analysis results showed that target genes were significantly enriched in cancer pathways,proteoglycan in cancer,human papillomavirus infection,breast cancer,regulatory stem cell pluripotency and other signaling pathways.miR-150 target gene interaction analysis results revealed that target genes PIK3R1,PIK3CB and CTNNB1 had more targeted relationships with other proteins,and involved in estrogen signaling pathway,cell senescence,phosphatidylinositol signaling system,TGF-beta and other pathways.The expression of miR-150 in different tissues and diseases was analyzed by YM500V2 online software,it was found that the expression level of miR-150 was the highest in blood,relatively high in ovary,pancreas,lymph and skin,and very low or no expression level in brain and adrenal cortex.These results would provide theoretical reference for investigating the function and regulatory mechanism of miR-150,and further laid the foundation for the regulatory mechanism of miR-150 in the ovarian development of sheep.

Key words: sheep; miR-150; target gene; bioinformatics

中图分类号:

付强, 岳巧娴, 宋鹏琰, 周荣艳. 绵羊miR-150靶基因预测及生物信息学分析[J]. 中国畜牧兽医, 2021, 48(7): 2333-2341.

FU Qiang, YUE Qiaoxian, SONG Pengyan, ZHOU Rongyan. Target Gene Prediction and Bioinformatics Analysis of miR-150 in Sheep[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(7): 2333-2341.

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