Inducible gene expression from the plastid genome by a synthetic riboswitch
Inducible gene expression from the plastid genome by a synthetic riboswitch
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DOI：
10.1073/pnas.0914423107
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发表时间：
2010-04-06
影响因子：
11.1
通讯作者：
Bock, Ralph
中科院分区：
文献类型：
--
作者：
Verhounig, Andreas;Karcher, Daniel;Bock, Ralph
核糖开关是一种天然的RNA传感器，可以调节基因的表达以响应配体的结合。核糖开关在原核生物和真核生物中已被发现，但在细胞器(线粒体和叶绿体)中未知。在这里，我们测试了为叶绿体设计核糖开关的可能性，这是一种在很大程度上依赖于基因表达的翻译控制的遗传系统。为此，我们使用了细菌核糖开关，并在硅胶中对其进行了修饰，以满足叶绿体中翻译调控的要求。然后，通过稳定转化烟草叶绿体基因组，在体内测试了这些工程开关的功能。我们报告了一种合成的核糖开关的鉴定，它作为外源应用的配体茶碱响应于其在质体中基因表达的有效翻译调节而发挥作用。这种核糖开关为叶绿体基因组工程提供了一种新的工具，促进了叶绿体基因和转基因的严格调控和诱导表达，因此在功能基因组学和生物技术中有广泛的应用。
Riboswitches are natural RNA sensors that regulate gene expression in response to ligand binding. Riboswitches have been identified in prokaryotes and eukaryotes but are unknown in organelles (mitochondria and plastids). Here we have tested the possibility to engineer riboswitches for plastids (chloroplasts), a genetic system that largely relies on translational control of gene expression. To this end, we have used bacterial riboswitches and modified them in silico to meet the requirements of translational regulation in plastids. These engineered switches were then tested for functionality in vivo by stable transformation of the tobacco chloroplast genome. We report the identification of a synthetic riboswitch that functions as an efficient translational regulator of gene expression in plastids in response to its exogenously applied ligand theophylline. This riboswitch provides a novel tool for plastid genome engineering that facilitates the tightly regulated inducible expression of chloroplast genes and transgenes and thus has wide applications in functional genomics and biotechnology.