Liu Qikun, Axtell Michael J
Department of Biology, Pennsylvania State University, University Park, PA, 16802, USA.
Methods Mol Biol. 2015;1284:287-303. doi: 10.1007/978-1-4939-2444-8_14.
microRNA (miRNA) mediated repression of target genes plays essential roles in a variety of functions in plants. An easy-to-use method that can effectively validate functional miRNA-target interactions in plants thus is of particular interest. Here, we describe an Agrobacterium tumefaciens-mediated in vivo assay utilizing a dual-luciferase reporter system. With this method, the strength of miRNA-mediated target repression can be quantified at both the mRNA (via qRT-PCR) and protein (via dual-luciferase assay) levels quickly and accurately.
微小RNA（miRNA）介导的靶基因抑制在植物的多种功能中起着至关重要的作用。因此，一种能够有效验证植物中功能性miRNA-靶标相互作用的简便方法备受关注。在此，我们描述了一种利用双荧光素酶报告系统的根癌农杆菌介导的体内分析方法。通过这种方法，可以在mRNA（通过qRT-PCR）和蛋白质（通过双荧光素酶分析）水平上快速准确地定量miRNA介导的靶标抑制强度。
