| Identification of RNA-binding Proteins by RNA Ligand-based cDNA Expression Library Screening
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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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| In situ Hybridization in Zebrafish Embryos
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Author:
Date:
2012-02-05
[Abstract] In situ hybridization is routinely used to examine the gene expression level and location of embryos. This protocol is modified from the Thisse protocol and is a detailed description of the in situ hybridization procedures in zebrafish embryos.
[摘要] 原位杂交常规用于检查胚胎的基因表达水平和位置。 该协议是从Thisse协议修改的，并且是斑马鱼胚胎中原位杂交程序的详细描述。
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| Preparation of Torula Yeast RNA for Hybe Solutions
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Author:
Date:
2012-02-05
[Abstract] In situ hybridization in zebrafish embryos frequently suffers from high background signal. Torular yeast RNA is often added to reduce the non-specific binding that leads to the high background signal. Without appropriate preparation, torular yeast RNA can also add background to the staining. This method is an easy and quick way to clean torular RNA for Hybe solution used in in situ hybridization.
[摘要] 原位杂交常规用于检查胚胎的基因表达水平和位置。 该协议是从Thisse协议修改的，并且是斑马鱼胚胎中原位杂交程序的详细描述。
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