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Solexa 深度测序技术鉴定和分析奶山羊(Capra hircus)乳腺中的 microRNA。

Identification and characterization of microRNA in the dairy goat (Capra hircus) mammary gland by Solexa deep-sequencing technology.

机构信息

Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, College of Animal Science and Veterinary Medicine, Shandong Agricultural University, 61 Daizong Avenue, Taian 271018, China.

出版信息

Mol Biol Rep. 2012 Oct;39(10):9361-71. doi: 10.1007/s11033-012-1779-5. Epub 2012 Jul 5.

Abstract

microRNAs (miRNAs) perform critical roles in various biological and metabolic processes by regulating gene expression at the post-transcriptional level. To investigate the functional roles of miRNAs in the lactating mammary gland of Capra hircus, a library was constructed from the lactating mammary glands of Laoshan dairy goats (C. hircus) during early lactation. The miRNA expression profiles were systematically screened, and miRNAs were identified and characterized using Solexa deep-sequencing technology and bioinformatics. As a result, a total of 18,031,615 clean reads were obtained representing 305,711 unique sRNAs. A total of 12,086,616 sRNAs representing 3,701 unique sRNAs matched the known Bos taurus miRNA precursors in miRBase 17.0, and 300 known miRNAs and 15 miRNA were discovered. In addition, 131 novel miRNAs sequences were also obtained, and 147,703 putative targets were predicted. GO and KEGG pathway analysis showed that the majority of targets were involved in cellular processes and metabolic pathways. The 290 known miRNAs, 14 miRNA and 38 novel miRNAs were validated by sequencing a second library that was constructed from the same tissues as the first library. Our study provided the first large-scale identification and characterization of miRNAs in the mammary gland tissue of the dairy goat. The results indicate that the regulation of miRNA-mediated gene expression occurs during early lactation in dairy goats. This study significantly enriches the C. hircus miRNA repertoire and provides a reference for the elucidation of complex miRNA-mediated regulatory networks for gene expression in the physiology and developmental progression of the lactating mammary gland.

摘要

微小 RNA(miRNAs)通过在转录后水平调节基因表达,在各种生物和代谢过程中发挥关键作用。为了研究 miRNAs 在 Capra hircus 泌乳乳腺中的功能作用,本研究从崂山奶山羊(C. hircus)哺乳期早期的泌乳乳腺中构建了一个文库。使用 Solexa 深度测序技术和生物信息学系统地筛选了 miRNA 表达谱,鉴定和描述了 miRNAs。结果共获得了 18,031,615 条清洁读数,代表 305,711 个独特的 sRNAs。总共 12,086,616 个 sRNAs 代表 3,701 个独特的 sRNAs 与 miRBase 17.0 中的已知 Bos taurus miRNA 前体匹配,发现了 300 个已知 miRNA 和 15 个 miRNA。此外,还获得了 131 个新的 miRNA 序列,预测了 147,703 个潜在靶标。GO 和 KEGG 通路分析表明,大多数靶标参与细胞过程和代谢途径。通过对与第一个文库相同组织构建的第二个文库进行测序,验证了 290 个已知 miRNA、14 个 miRNA 和 38 个新 miRNA。本研究首次大规模鉴定和描述了奶山羊乳腺组织中的 miRNAs。结果表明,miRNA 介导的基因表达调控发生在奶山羊哺乳期早期。该研究显著丰富了 C. hircus miRNA 库,为阐明泌乳乳腺生理和发育过程中 miRNA 介导的基因表达调控复杂网络提供了参考。

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