基因选和小分子的选择基于合成的核糖开关,激活蛋白质翻译
Shawn K Desai1, Justin P Gallivan
1Department of Chemistry and Center for Fundamental and Applied Molecular Evolution, Emory University, 1515 Dickey Drive, Atlanta, Georgia 30322, USA.
Journal of the American Chemical Society
|October 14, 2004
概括
合成的 рибо开关能够对大肠杆菌中的小分子进行敏感的遗传选择. 这一突破使得从突变库中发现了具有新型连接体特异性的新核突变开关.
科学领域:
- 合成生物学 合成生物学
- 分子生物学分子生物学
- 生物化学 生物化学
背景情况:
- 基因选择对生物分子选具有强大作用,但对非内源性小分子在体内具有挑战性.
- 现有的方法通常依赖于难以设计的小分子蛋白相互作用,并且可以通过宿主生物的特性 (如酵母的缓慢生长) 来限制.
研究的目的:
- 为了证明合成 рибо开关在体内基因查和选择小分子中的实用性.大肠杆菌.
- 为了证明在体外选择的aptamer特性转化为体内遗传选择.
- 探索发现具有新的连接体特异性的新型核糖开关的潜力.
主要方法:
- 工程合成 рибо开关,激活蛋白质翻译,以响应特定的小分子.
- 利用大肠杆菌作为宿主系统进行体内查和选择.
- 从大型突变库中进行了选择实验,以扩大具有所需的核糖开关特异性的细胞.
主要成果:
- 合成 рибо开关成功实现了敏感的基因选和对大肠杆菌中小分子的选择.
- 在体外选择的体的分子歧视是直接适用于体内遗传选择系统.
- 证明了具有特定 рибо开关连接体特异性的细胞的选择性放大,来自具有相关特异性的细胞的百万倍过量.
结论:
- 合成的 рибо开关是有效的工具,用于E. coli中小分子的体内遗传选择.
- 该系统允许发现具有量身定制的连接体特异性的新型核糖开关.
- 该方法通过遗传选择促进了分子相互作用和酶工程的探索.
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