模板超卷由酵母 GAL4 蛋白和菌体 T7 RNA 聚合酶的仿真体进行
E A Ostrander1, P Benedetti, J C Wang
1Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, MA 02138.
概括
研究人员通过将酵母 GAL4 DNA 结合域与菌体 T7 RNA 聚合酶融合而创建了一个仿真酶. 这种工程蛋白在转录过程中专门超级卷结含有T7促进子序列的DNA.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 遗传学 是一个遗传学.
背景情况:
- 菌体T7RNA聚合酶是转录的关键酶.
- 酵母 GAL4 蛋白质有一个 DNA 结合域,可以与 DNA 结合.
研究的目的:
- 创建一个结合T7RNA聚合酶和GAL4DNA结合域的嵌合酶.
- 为了研究这种工程酶的DNA超卷能力.
主要方法:
- 酵母 GAL4 DNA 结合域与细菌 T7 RNA 聚合酶的基因融合.
- 在体外和体外转录试验中使用化学酶进行了转录试验.
- 通过化学聚合酶诱导的DNA超级卷的分析.
主要成果:
- 嵌合体酶保留了两个父蛋白的特性.
- GAL4结合促进了DNA的定,导致超的DNA循环形成.
- 超级卷发生在转录过程中,取决于T7促进子序列.
结论:
- 嵌合体聚合酶特别超级卷曲含有T7促进体的DNA段.
- 这种工程酶作为一种新的工具,用于在体外和体内的向DNA操纵.
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