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基于高通量测序的亚麻 microRNA 基因及其靶基因的鉴定与分析:亚麻 miRNA 基因的鉴定。

Genome-wide identification and characterization of microRNA genes and their targets in flax (Linum usitatissimum): Characterization of flax miRNA genes.

机构信息

Biochemical Sciences Division, CSIR-National Chemical Laboratory, Pune, 411008, India.

出版信息

Planta. 2013 Apr;237(4):1149-61. doi: 10.1007/s00425-012-1833-5. Epub 2013 Jan 6.

DOI:10.1007/s00425-012-1833-5
PMID:23291876
Abstract

MicroRNAs (miRNAs) are small (20-24 nucleotide long) endogenous regulatory RNAs that play important roles in plant growth and development. They regulate gene expression at the post-transcriptional level by translational repression or target degradation and gene silencing. In this study, we identified 116 conserved miRNAs belonging to 23 families from the flax (Linum usitatissimum L.) genome using a computational approach. The precursor miRNAs varied in length; while most of the mature miRNAs were 21 nucleotide long, intergenic and showed conserved signatures of RNA polymerase II transcripts in their upstream regions. Promoter region analysis of the flax miRNA genes indicated prevalence of MYB transcription factor binding sites. Four miRNA gene clusters containing members of three phylogenetic groups were identified. Further, 142 target genes were predicted for these miRNAs and most of these represent transcriptional regulators. The miRNA encoding genes were expressed in diverse tissues as determined by digital expression analysis as well as real-time PCR. The expression of fourteen miRNAs and nine target genes was independently validated using the quantitative reverse transcription PCR (qRT-PCR). This study suggests that a large number of conserved plant miRNAs are also found in flax and these may play important roles in growth and development of flax.

摘要

微 RNA(miRNA)是小的(20-24 个核苷酸长)内源性调节 RNA,在植物生长和发育中发挥重要作用。它们通过翻译抑制或靶降解和基因沉默来调节转录后水平的基因表达。在这项研究中,我们使用计算方法从亚麻(Linum usitatissimum L.)基因组中鉴定出 116 个属于 23 个家族的保守 miRNA。前体 miRNA 的长度不同;大多数成熟 miRNA 长 21 个核苷酸,位于基因间区,并在其上游区域显示 RNA 聚合酶 II 转录物的保守特征。亚麻 miRNA 基因的启动子区域分析表明 MYB 转录因子结合位点的普遍性。鉴定出四个包含三个系统发育群成员的 miRNA 基因簇。此外,为这些 miRNA 预测了 142 个靶基因,其中大多数代表转录调节剂。通过数字表达分析和实时 PCR 确定了 miRNA 编码基因在不同组织中的表达。使用定量逆转录 PCR(qRT-PCR)独立验证了 14 个 miRNA 和 9 个靶基因的表达。本研究表明,大量保守的植物 miRNA 也存在于亚麻中,这些 miRNA 可能在亚麻的生长和发育中发挥重要作用。

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