Department of Life Science, National Taiwan University, Taipei 10617, Taiwan.
Institute of Plant Biology, National Taiwan University, Taipei 10617, Taiwan.
ACS Synth Biol. 2023 Apr 21;12(4):1320-1330. doi: 10.1021/acssynbio.3c00066. Epub 2023 Mar 30.
As the demand for sustainable energy has increased, photoautotrophic cyanobacteria have become a popular platform for developing tools in synthetic biology. Although genetic tools are generally available for several model cyanobacteria, such tools have not yet been developed for many other strains potentially suitable for industrial applications. Additionally, most inducible promoters in cyanobacteria are controlled by chemical compounds, but adding chemicals into growth media on an industrial scale is neither cost-effective nor environmentally friendly. Although using light-controlled promoters is an alternative approach, only a cyanobacterial expression system inducible by green light has so far been described and employed for such applications. In this study, we have established a conjugation-based technique to express a reporter gene () in the nonmodel cyanobacterium, PCC 9212. We also identified a promoter specifically activated by far-red light from the Far-Red Light Photoacclimation gene cluster of sp. JSC-1. This promoter, , was successfully used to drive expression. is tightly regulated by light quality (i.e., wavelength) and leads to an approximately 30-fold increase in EYFP production when cells were exposed to far-red light. The induction level was controlled by the far-red light intensity, and induction stopped when cells were returned to visible light. This system has the potential for further applications in cyanobacteria by providing an additional choice of light wavelength to control gene expression. Collectively, this study developed a functional gene-expression system for PCC 9212 that can be regulated by exposing cells to far-red light.
随着对可持续能源的需求增加,光自养蓝藻已成为开发合成生物学工具的热门平台。尽管有几种模式蓝藻可使用一般的遗传工具,但许多其他可能适用于工业应用的菌株尚未开发出这些工具。此外,蓝藻中的大多数诱导型启动子受化学化合物控制,但在工业规模上将化学物质添加到生长培养基中既不经济也不环保。虽然使用光控启动子是一种替代方法,但迄今为止,仅描述并应用了一种可被绿光诱导的蓝藻表达系统来实现这种应用。在这项研究中,我们建立了一种基于接合的技术,可在非模式蓝藻 PCC 9212 中表达报告基因()。我们还鉴定了一个特定的启动子,该启动子由 sp. JSC-1 的远红光光适应基因簇中的远红光激活。这个启动子,,成功地用于驱动表达。受光质(即波长)的严格调控,当细胞暴露于远红光时光诱导可使 EYFP 的产量增加约 30 倍。诱导水平受远红光强度的控制,当细胞返回可见光时诱导停止。该系统通过提供控制基因表达的另一种波长的选择,为蓝藻的进一步应用提供了潜力。总之,本研究为 PCC 9212 开发了一种可通过暴露细胞于远红光来调节的功能基因表达系统。