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全球运行测序测量Saccharomyces cerevisiae中的新生转录

Michael J O'Brien¹, Katherine Gurdziel², Athar Ansari¹

¹Department of Biological Science, Wayne State University, Detroit, MI 48202, USA.

Methods (San Diego, Calif.)

|June 25, 2023

概括

全球运行测序 (GRO-seq) 现在提供了一种强大的方法,用于绘制初芽酵母的新生转录. 这种技术精确地识别了整个基因组中的活跃基因和转录动态.

科学领域:

分子生物学分子生物学
基因组学就是基因组学.
酵母遗传学酵母遗传学

背景情况:

全球运行测序 (GRO-seq) 是用于全基因组新生转录分析的验证方法.
之前的GRO-seq应用已经在各种模型生物中取得了成功,包括人类,植物和.
缺乏针对芽生长酵母 (Saccharomyces cerevisiae) 的特定协议.

研究的目的:

为芽酵母 (Saccharomyces cerevisiae) 适应和描述一个全球运行测序 (GRO-seq) 协议.
为了使这一重要的模型生物体中新生的转录能够进行全基因组分析.
为细致研究酵母转录调节提供一个工具.

主要方法:

开发了一个针对Saccharomyces cerevisiae量身定制的GRO-seq协议.
涉及透活跃生长的酵母细胞,以允许现场转录.
包括化UTP,随后是标记新生的RNA的亲和性净化,cDNA库的准备,深度测序和基因组映射.

主要成果:

该协议成功地在整个酵母基因组中映射了转录活性RNA聚合酶的位置.
能够在特定条件下识别所有活跃转录的基因.
提供关于RNA聚合酶延长率,终结效率和促进子方向性的全基因组数据.

结论:

这种适应的GRO-seq协议是有效的研究全球转录在芽酵母.
该方法对于识别短暂,不稳定的转录有价值.
促进了对转录动态的全面分析,包括Saccharomyces cerevisiae中的基因活性,延长和终止.

关键词:

新生的RNARNA是什么意思的RNA聚合酶.测序测序是指测序的时间.转录开始运行转录的定向性转录的定向性转录启动的开始.转录终止转录终止

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