牛乳外泌体中miRNA的测序与分析

Sequencing and Analysis of MicroRNAs in Bovine Milk Exosomes

采用密度梯度离心法提取牛乳外泌体,利用Illumina测序技术对外泌体中非编码小RNA(sRNA)进行测序,探索牛乳microRNA(miRNA)的表达谱。对原始序列进行质量控制,共获得3 899 629条纯净sRNA序列,长度集中于28 nt,与数据库比对后鉴定到61种已知miRNA,346种新miRNA。基因本体论富集结果表明,牛乳外泌体miRNA在细胞过程、单一生物体过程、代谢过程等生物过程发挥作用;主要构成细胞、细胞器等组成;主要参与结合、催化活性、转运蛋白活性等分子功能。京都基因与基因组百科全书通路富集结果表明,已知miRNA与新miRNA靶基因均显著富集在百日咳(ko05133)、趋化因子信号通路(ko04062)、内吞作用(ko04144)、溶酶体(ko04142)等通路,牛乳外泌体miRNA在特定信号通路中发挥重要作用。

In this study,density-gradient centrifugation was used to extract bovine milk exosomes,and the small non-coding RNA(sRNA)in the exosomes were sequenced by Illumina sequencing technology to explore the expression profile of bovine milk microRNAs(miRNAs).Through quality control of the original sequence,a total of 3899629 pure sRNA sequences were obtained,and their length was concentrated at 28 nt.By comparison with the database,61 known miRNAs and 346 novel miRNAs were identified.The results of gene ontology enrichment analysis showed that miRNAs from bovine milk exosomes played a critical role in various biological processes such as cell process,single organism process and metabolic process.It mainly consisted of cells and organelles and was mainly involved in molecular functions such as binding,catalytic activity and transporter activity.The results of Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis showed that the known miRNAs and the new miRNA target genes were significantly enriched in pertussis(ko05133),chemokine signal pathway(ko04062),endocytosis(ko04144),lysosome(ko04142)and other pathways,and bovine milk exosome miRNAs played an important role in specific signaling pathways.