miR-192进化分析、靶基因预测及组织表达分析

Evolutionary Analysis,Target Gene Prediction and Tissue Expression Analysis of miR-192

【目的】研究miR-192的生物进化进程和潜在靶基因功能,以及miR-192在大围子猪(脂肪型猪)和大约克夏猪(瘦肉型猪)脂肪组织中的差异表达。【方法】利用miBase数据库获取miR-192成熟序列,并利用Ensembl数据库定位miR-192在不同物种基因组的分布特征;采用Mega 11.0软件程序构建miR-192系统进化树;通过实时荧光定量PCR检测miR-192在大围子猪和大约克夏猪背部脂肪组织中的表达情况,并比较表达差异;利用不同在线网站预测猪miR-192靶基因,并对靶基因进行GO功能和KEGG通路富集分析。【结果】miR-192在生物进化过程中具有高度保守性;实时荧光定量PCR结果显示,miR-192在大围子猪背部脂肪组织的表达量极显著高于大约克夏猪(P<0.01);GO功能和KEGG通路富集分析结果表明,miR-192靶基因主要起转录前调节和结合的作用,且主要在染色质和细胞核中发挥作用,多数基因富集在与脂肪代谢相关的信号通路;对3个网站不同物种预测结果取交集共获得猪miR-192保守靶基因5个。【结论】miR-192在生物进化过程中具有高度保守性,可能通过靶向调节靶基因的表达,从而在猪脂肪组织分化中起作用,为进一步探讨miR-192在脂肪组织分化中的调控机制提供理论基础。

【Objective】The purpose of this study was to investigate the biological evolutionary process and potential target gene function of miR-192,and the expression differences of miR-192 in adipose tissue of Daweizi pigs(obese-type)and Yorkshire(lean-type).【Method】miR-192 mature sequences were obtained using the miBase database,and the distribution characteristics of miR-192 on the genomes of different species was located using the Ensembl database.The phylogenetic tree of miR-192 was constructed using the Mega 11.0.The expression of miR-192 in the dorsal adipose tissue of Daweizi pigs and Yorkshire were analysised by quantitative Real-time PCR,and the expression differences were compared.Multiple target gene prediction sites were used to predict the target genes that might bind to pig miR-192,and GO function and KEGG enrichment analysis were performed on the target genes.【Result】miR-192 maintained a high degree of conservation during biological evolution.Quantitative Real-time PCR results showed that the expression of miR-192 in the dorsal adipose tissue of Daweizi pig was extremely significantly higher than that of Yorkshire(P<0.01).The results of GO function and KEGG pathway enrichment showed that the target gene of miR-192 mainly play the role of pre-transcriptional regulation and binding,and mainly function in chromatin and nucleus,and most genes were enriched in signaling pathways related to adipose metabolism.Five conserved target genes of pig miR-192 were obtained from the intersection of prediction results of different species on three websites.【Conclusion】miR-192 was highly conserved in biological evolution and might regulate the expression of target genes by targeting,and played an important role in adipose tissue differentiation of pigs,which provided a theoretical basis for further research on the regulatory mechanism of miR-192 in adipose tissue differentiation.