Fachbereich Biologie, Synthetische Biologie, Technische Universität Darmstadt, Schnittspahnstr. 10, 64287 Darmstadt, Germany.
Microbiology (Reading). 2013 Jul;159(Pt 7):1416-1422. doi: 10.1099/mic.0.067322-0. Epub 2013 May 15.
We have demonstrated the portability of theophylline-dependent synthetic riboswitches for the conditional control of gene expression in Streptomyces coelicolor. The riboswitches mediate dose-dependent, up to 260-fold activation of reporter gene expression. Riboswitch regulation is a simple method requiring a sequence of only ~85 nt to be inserted between a transcriptional start site and the start codon; no additional auxiliary factors are necessary. The promoters galP2, ermEp1 and SF14 worked well in concert with the riboswitches. They allowed theophylline-dependent expression of not only the heterologous β-glucuronidase reporter gene but also dagA, an endogenous agarase gene. The successful combination of all tested promoters with the riboswitches underlines the orthogonality of riboswitch regulation. We anticipate that any additional natural or synthetic promoters can be combined with the riboswitch.
我们已经证明了茶碱依赖性合成核糖体开关在链霉菌中条件控制基因表达的可转移性。这些核糖体开关介导了依赖于剂量的、高达 260 倍的报告基因表达激活。核糖体开关调控是一种简单的方法,只需要在转录起始位点和起始密码子之间插入大约 85 个核苷酸的序列,不需要额外的辅助因子。启动子 galP2、ermEp1 和 SF14 与核糖体开关协同作用良好。它们不仅允许茶碱依赖性表达异源β-葡萄糖醛酸酶报告基因,还允许内源性琼脂酶基因 dagA 的表达。所有测试的启动子与核糖体开关的成功组合突出了核糖体开关调控的正交性。我们预计任何额外的天然或合成启动子都可以与核糖体开关结合。