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Author:
Date:
2016-01-20
[Abstract] We previously reported when a portion of the Requiem (REQ/DPF2) messenger ribonucleic acid (mRNA) 3’ untranslated region (3’UTR), referred to as G8, was overexpressed in K562 cells, β-globin expression was induced, suggesting that the 3’UTR of REQ mRNA plays a physiological role (Kim et al., 2014). To identify trans-acting factors that bind to the REQ 3’UTR, we describe the RNA ligand based cDNA expression library screening method. This protocol could be adapted to detect specific RNA-protein interactions. Following this method, we identified six positive clones in the initial round of screening and four pure clones after sib-screening. This protocol was originally published in Kim et al. (2014).
[摘要] 我们先前报道当在K562细胞中过表达Requiem（REQ/DPF2）信使核糖核酸（mRNA）3'非翻译区（3'UTR），称为G8的一部分时，诱导β-球蛋白表达， REQ mRNA的3'UTR起生理作用（Kim等人，2014）。 为了鉴定结合REQ 3'UTR的反式作用因子，我们描述了基于RNA配体的cDNA表达文库筛选方法。 这个协议可以适应于检测特定的RNA-蛋白相互作用。 按照这种方法，我们在初始轮筛选中鉴定了6个阳性克隆，在同胞筛选后鉴定了4个纯克隆。 该协议最初发表于Kim等人（2014）。
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