来自酵母的DNA结合蛋白的净化和克隆,该蛋白结合于声器和激活元件
Cell
|December 4, 1987
概括
研究人员净化了RAP1 (抑制剂激活蛋白1),这是一种参与酵母基因调节的DNA结合蛋白. 它的基因破坏是致命的,RAP1作为转录调节器,影响基因激活和抑制.
科学领域:
- 分子生物学分子生物学
- 酵母遗传学 酵母遗传学
- 转录条例 转录条例 转录条例
背景情况:
- 一种DNA结合蛋白,RAP1 (以前称为SBF-E),与酵母中的调节性DNA位点相互作用.
- RAP1没有被已知的静音位点调节器编码.
- 关于RAP1在基因调节中的确切功能尚未完全阐明.
研究的目的:
- 为了净化RAP1蛋白并分离其基因.
- 研究RAP1在转录调节中的作用.
- 为了确定RAP1是否可以在基因激活和抑制中发挥作用.
主要方法:
- 用于蛋白质净化的DNA亲和色谱.
- 使用抗体进行基因隔离的Lambda gt11基因组图书馆查.
- 基因干扰以评估致命性和体内功能.
- 在体外结合测试以确定DNA点.
- 在体内测试以评估调节部位的功能替代.
主要成果:
- 净化了RAP1并将其基因分离出来.
- 在酵母中,RAP1基因的破坏是致命的.
- RAP1与MATα和核糖体蛋白基因的上游激活部位 (UAS) 结合.
- 在体内,与UAS结合的RAP1位点可以功能性地取代沉声器结合的位点.
结论:
- RAP1 是酵母中一个必不可少的基因.
- RAP1是一种多功能转录调节器.
- RAP1的功能 (激活或抑制) 取决于其结合点的上下文.
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