Regulation of Gene Expression in Diverse Cyanobacterial Species by Using Theophylline-Responsive Riboswitches
Regulation of Gene Expression in Diverse Cyanobacterial Species by Using Theophylline-Responsive Riboswitches
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DOI：
10.1128/aem.01697-14
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发表时间：
2014-11-01
影响因子：
4.4
通讯作者：
Golden, James W.
中科院分区：
文献类型：
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作者：
Ma, Amy T.;Schmidt, Calvin M.;Golden, James W.
蓝细菌是目前正在开发为生物生产平台的光合细菌。它们从光中获取能量，从大气中的二氧化碳中获取碳，有些物种可以固定大气中的氮。开发蓝细菌用于生物燃料和其他生物产品的可再生生产的优势之一是它们易于基因操作，促进生物工程和合成生物学。为了扩展目前可用的遗传工具包，我们证明了合成茶碱响应核糖开关在四种不同蓝藻物种（包括最近分离的两种）中有效调控基因表达的效用。我们评估了一组六个核糖开关，它们驱动细长聚球藻 PCC 7942、Leptolyngbya sp 中黄色荧光蛋白报告基因的表达。菌株 BL0902，鱼腥藻属。菌株 PCC 7120 和集胞藻属 sp。菌株 WHSyn。我们证明核糖开关可以提供优于常用的 lacI(q)-P-trc 启动子系统的异丙基-β-D-硫代半乳糖苷诱导的基因表达调节。我们还表明，有毒蛋白 SacB 的表达可以得到有效调节，证明了核糖开关调节不利于生物量积累的蛋白质的效用。总而言之，这项工作的结果证明了核糖开关在多种蓝细菌的基因工程和合成生物学背景下的实用性和易用性，这将促进藻类生物技术的发展。
Cyanobacteria are photosynthetic bacteria that are currently being developed as biological production platforms. They derive energy from light and carbon from atmospheric carbon dioxide, and some species can fix atmospheric nitrogen. One advantage of developing cyanobacteria for renewable production of biofuels and other biological products is that they are amenable to genetic manipulation, facilitating bioengineering and synthetic biology. To expand the currently available genetic toolkit, we have demonstrated the utility of synthetic theophylline-responsive riboswitches for effective regulation of gene expression in four diverse species of cyanobacteria, including two recent isolates. We evaluated a set of six riboswitches driving the expression of a yellow fluorescent protein reporter in Synechococcus elongatus PCC 7942, Leptolyngbya sp. strain BL0902, Anabaena sp. strain PCC 7120, and Synechocystis sp. strain WHSyn. We demonstrated that riboswitches can offer regulation of gene expression superior to that of the commonly used isopropyl-beta-D-thiogalactopyranoside induction of a lacI(q)-P-trc promoter system. We also showed that expression of the toxic protein SacB can be effectively regulated, demonstrating utility for riboswitch regulation of proteins that are detrimental to biomass accumulation. Taken together, the results of this work demonstrate the utility and ease of use of riboswitches in the context of genetic engineering and synthetic biology in diverse cyanobacteria, which will facilitate the development of algal biotechnology.