National Center for Molecular Genetics and Breeding of Animal, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, No. 2 Yuanmingyuan West Road, Haidian District, Beijing 100193, People's Republic of China.
Mol Biol Rep. 2011 Jun;38(5):3161-71. doi: 10.1007/s11033-010-9987-3. Epub 2010 Feb 7.
Ovis aries is one of the most important agricultural livestock for meat production, and also is an ideal model organism for biological and comparative genomics studies. Many miRNAs have been reported for their important roles in developmental processes in various animals, but there is limited information about O. aries miRNAs. In this study, combining a computational method based on expressed sequence tag (EST) analysis with experimental identification based on small RNA cDNA library, we identified 31 miRNAs belong to 24 families in sheep, 2 of which were novel miRNAs which had never been previously identified in any species. Especially, we cloned 12 miRNAs from the sheep skeletal muscle, which were good candidate miRNAs to be studied about the miRNA-dependant regulated process of muscle development, and we identified four pairs of miRNA/miRNA and one pair of miRNA-3p/miRNA-5p from sheep EST sequences. Expression analysis indicated that some miRNAs were expressed in a specific tissue, and the pair of miRNA-3p/miRNA-5p and one pair of miRNA/miRNA had a similar relative expression pattern in some tissues, respectively. Further, we predicted 120 potential target genes of 31 oar-miRNAs on the 3'UTR of O. aries genes. Gene ontology analysis showed that most of these genes took part in the cellular process and metabolic process. Our results enriched the O. aries miRNA database and provided useful information for investigating biological functions of miRNAs and miRNA in sheep.
绵羊是重要的肉用农业家畜之一,也是生物学和比较基因组学研究的理想模式生物。许多 miRNA 在各种动物的发育过程中发挥着重要作用,已有相关报道,但关于绵羊 miRNAs 的信息却十分有限。本研究采用基于表达序列标签(EST)分析的计算方法和基于小 RNA cDNA 文库的实验鉴定方法,在绵羊中鉴定出了 31 个属于 24 个家族的 miRNA,其中 2 个是新的 miRNA,在任何物种中都从未被鉴定过。特别是,我们从绵羊骨骼肌中克隆了 12 个 miRNA,它们是研究 miRNA 调控肌肉发育过程的候选 miRNA,我们从绵羊 EST 序列中鉴定出了 4 对 miRNA/miRNA 和 1 对 miRNA-3p/miRNA-5p。表达分析表明,一些 miRNA 在特定组织中表达,并且 miRNA-3p/miRNA-5p 对和 miRNA/miRNA 对在某些组织中具有相似的相对表达模式。此外,我们预测了 31 个 oar-miRNAs 在 O. aries 基因 3'UTR 上的 120 个潜在靶基因。GO 分析表明,这些基因大多参与细胞过程和代谢过程。我们的研究结果丰富了绵羊 miRNA 数据库,为研究 miRNA 在绵羊中的生物学功能提供了有用的信息。