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Isolation and identification of miRNAs in Jatropha curcas

机译:麻疯树中miRNA的分离与鉴定

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MicroRNAs (miRNAs) are small noncoding RNAs that play crucial regulatory roles by targeting mRNAs for silencing. To identify miRNAs in Jatropha curcas L, a bioenergy crop, cDNA clones from two small RNA libraries of leaves and seeds were sequenced and analyzed using bioinformatic tools. Fifty-two putative miRNAs were found from the two libraries, among them six were identical to known miRNAs and 46 were novel. Differential expression patterns of 15 miRNAs in root, stem, leave, fruit and seed were detected using quantitative real-time PCR. Ten miRNAs were highly expressed in fruit or seed, implying that they may be involved in seed development or fatty acids synthesis in seed. Moreover, 28 targets of the isolated miRNAs were predicted from a jatropha cDNA library database. The miRNA target genes were predicted to encode a broad range of proteins. Sixteen targets had clear BLASTX hits to the Uniprot database and were associated with genes belonging to the three major gene ontology categories of biological process, cellular component, and molecular function. Four targets were identified for JcumiR004. By silencing JcumiR004 primary miRNA, expressions of the four target genes were up-regulated and oil composition were modulated significantly, indicating diverse functions of JcumiR004.
机译:MicroRNA(miRNA)是小的非编码RNA,通过靶向沉默的mRNA发挥关键的调节作用。为了鉴定麻疯树麻风树(一种生物能源作物)中的miRNA,对来自叶子和种子的两个小RNA库的cDNA克隆进行了测序,并使用生物信息学工具进行了分析。从两个文库中发现了52个推定的miRNA,其中六个与已知的miRNA相同,而46个是新颖的。使用实时定量PCR检测了15种miRNA在根,茎,叶,果实和种子中的差异表达模式。十个miRNA在果实或种子中高表达,这暗示它们可能参与种子的发育或种子中的脂肪酸合成。此外,从麻风树属cDNA文库数据库预测了分离的miRNA的28个靶标。预测miRNA靶基因可编码多种蛋白质。 16个靶标在Uniprot数据库中具有明确的BLASTX命中率,并且与属于生物过程,细胞成分和分子功能的三个主要基因本体论类别的基因相关联。为JcumiR004确定了四个目标。通过沉默JcumiR004初级miRNA,四个靶基因的表达被上调并且油的成分被显着调节,表明JcumiR004的功能多样。

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