A simplified vector system for visualization of localized RNAs in Schizosaccharomyces pombe
A simplified vector system for visualization of localized RNAs in Schizosaccharomyces pombe
复制标题
用于可视化裂殖酵母中局部 RNA 的简化载体系统
DOI：
10.1080/09168451.2016.1158633
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发表时间：
2016
期刊：
影响因子：
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通讯作者：
Nobuyuki Terouchi and Tokio Tani
中科院分区：
文献类型：
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作者：
Tomoko Takeuchi-Andoh;Sayaka Ohba;Yu Shinoda;Ayako Fuchita;Sachiko Hayashi;Emi Nishiyoshi;Nobuyuki Terouchi and Tokio Tani
RNA定位是细胞极化和分化所必需的重要事件。虽然目前已有多种方法用于检测定位的RNA，但对于裂殖酵母还没有开发出一种简化的检测系统。在本研究中，我们描述了一种新的用于局部RNA INS可视化的载体系统。使用U1a-Tag-GFP系统。构建了表达U1a标记RNA的pREP1-U1a-tag载体和表达U1a-GFP融合蛋白的pREP2-U1AGFP表达载体。由于U1a-GFP蛋白与U1a标记的RNA结合，因此在表达这两种标记的细胞中都能在U1a标记的RNA位置观察到荧光。利用该系统成功地检测了U3 snoRNA的核仁定位，并通过筛选定位的RNA发现了定位于细胞核DNA区域的新RNA。该系统的建立将加速定位RNAs INS的研究。庞贝。
RNA localization is an important event that is essential for the polarization and differentiation of a cell. Although several methods are currently used to detect localized RNAs, a simplified detection system has not yet been developed forSchizosaccharomyces pombe. In the present study, we describe a new vector system for the visualization of localized RNAs inS. pombeusing a U1A-tag-GFP system. A pREP1-U1A-tag vector plasmid to express U1A-tagged RNA and a pREP2-U1AGFP plasmid to produce a U1A-GFP fusion protein were constructed for this system. Since the U1A-GFP protein binds U1A-tagged RNA, fluorescence is observed at the location of U1A-tagged RNA in cells expressing both of these. The nucleolar localization of U3 snoRNA was successfully detected using this system, and a novel RNA localized at the DNA region of the nucleus was found by screening localized RNAs. This system will accelerate the study of localized RNAs inS. pombe.