Modular Riboswitch Toolsets for Synthetic Genetic Control in Diverse Bacterial Species

详细信息    查看全文

• 作者：Christopher J. Robinson ; Helen A. Vincent ; Ming-Cheng Wu ; Phillip T. Lowe ; Mark S. Dunstan ; David Leys ; Jason Micklefield
• 刊名：Journal of the American Chemical Society
• 出版年：2014
• 出版时间：July 30, 2014
• 年：2014
• 卷：136
• 期：30
• 页码：10615-10624
• 全文大小：550K
• ISSN：1520-5126

文摘

Ligand-dependent control of gene expression is essential for gene functional analysis, target validation, protein production, and metabolic engineering. However, the expression tools currently available are difficult to transfer between species and exhibit limited mechanistic diversity. Here we demonstrate how the modular architecture of purine riboswitches can be exploited to develop orthogonal and chimeric switches that are transferable across diverse bacterial species, modulating either transcription or translation, to provide tunable activation or repression of target gene expression, in response to synthetic non-natural effector molecules. Our novel riboswitch鈥搇igand pairings are shown to regulate physiologically important genes required for bacterial motility in Escherichia coli and cell morphology in Bacillus subtilis. These findings are relevant for future gene function studies and antimicrobial target validation, while providing new modular and orthogonal regulatory components for deployment in synthetic biology regimes.