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Small RNA analysis in Petunia hybrida identifies unusual tissue-specific expression patterns of conserved miRNAs and of a 24mer RNA

机译:矮牵牛中的小RNA分析鉴定了保守的miRNA和24mer RNA的异常组织特异性表达模式

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摘要

Two pools of small RNAs were cloned from inflorescences of Petunia hybrida using a 5′-ligation dependent and a 5′-ligation independent approach. The two libraries were integrated into a public website that allows the screening of individual sequences against 359,769 unique clones. The library contains 15 clones with 100% identity and 53 clones with one mismatch to miRNAs described for other plant species. For two conserved miRNAs, miR159 and miR390, we find clear differences in tissue-specific distribution, compared with other species. This shows that evolutionary conservation of miRNA sequences does not necessarily include a conservation of the miRNA expression profile. Almost 60% of all clones in the database are 24-nucleotide clones. In accordance with the role of 24mers in marking repetitive regions, we find them distributed across retroviral and transposable element sequences but other 24mers map to promoter regions and to different transcript regions. For one target region we observe tissue-specific variation of matching 24mers, which demonstrates that, as for 21mers, 24mer concentrations are not necessarily identical in different tissues. Asymmetric distribution of a putative novel miRNA in the two libraries suggests that the cloning method can be selective for the representation of certain small RNAs in a collection.
机译:使用5'-连接依赖性和5'-连接依赖性方法从矮牵牛的花序中克隆了两个小RNA池。这两个库被整合到一个公共网站中,该网站允许针对359,769个独特克隆筛选单个序列。该文库包含15个具有100%同一性的克隆和53个与其他植物物种中描述的miRNA不匹配的克隆。对于两个保守的miRNA,miR159和miR390,我们发现与其他物种相比,组织特异性分布存在明显差异。这表明miRNA序列的进化保守性不一定包括miRNA表达谱的保守性。数据库中几乎所有克隆中的60%是24个核苷酸的克隆。根据24mers在标记重复区域中的作用,我们发现它们分布在逆转录病毒和转座因子序列中,而其他24mers则映射到启动子区域和不同的转录区域。对于一个靶区域,我们观察到匹配的24mer的组织特异性变异,这表明,对于21mers,24mer浓度在不同组织中不一定相同。推定的新型miRNA在两个文库中的不对称分布表明,克隆方法可以选择性地代表集合中某些小RNA的表达。

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