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Company: Sigma-Aldrich
Catalog#: 613304
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Detecting the Interaction of Double-stranded RNA Binding Protein, Viral Protein and Primary miRNA Transcript by Co-immunoprecipitation in planta
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2018-05-05
[Abstract]  MicroRNAs (miRNAs) play important roles in plant growth, development, and response to infection by microbes. Double-stranded RNA binding protein 1 (DRB1) facilitates the processing of primary miRNA transcripts into mature miRNAs. Recently, we found that NS3 protein encoded by rice stripe virus (RSV) associates with DRB1 and promotes miRNA biogenesis during RSV infection (Zheng et al., 2017). RNA co-immunoprecipitation (RIP) method was applied to identity association patterns among DRB1, NS3, and miRNA transcript. [摘要]  微小RNA(miRNA)在植物生长,发育和微生物感染反应中发挥重要作用。 双链RNA结合蛋白1(DRB1)有助于将初级miRNA转录物加工成成熟的miRNA。 最近,我们发现水稻条纹病毒(RSV)编码的NS3蛋白与DRB1相关并促进RSV感染期间的miRNA生物合成(Zheng等人,2017)。 RNA共免疫沉淀(RIP)方法被用于鉴定DRB1,NS3和miRNA转录物之间的关联模式。

【背景】在双链RNA(dsRNA)结合蛋白HYPONASTIC LEAVES1(DRB1 / HYL1)的帮助下,通过RNA酶III酶DICER-LIKE1(DCL1)从其初级转录物(pri-miRNA) 锌指蛋白SERRATE(SE)。 水稻条纹病毒(RSV)感染广泛地干扰miRNA积累。 我们发现RSV编码的非结构蛋白3(NS3)通过与水稻中的DRB1相互作用下调pri-miRNAs来促进miRNA积累(Zheng等人,2017)。 为了揭示NS3如何增强pri-miRNA的加工,我们使用免疫共沉淀(Co-IP)来说明NS3,DRB1和pri-miRNA体内的关系。 该协议有助于了解两种蛋白质和一种RNA转录本之间的关联模式。

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