Kelly Ciarán L, Taylor George M, Hitchcock Andrew, Torres-Méndez Antonio, Heap John T
Imperial College Centre for Synthetic Biology, Department of Life Sciences , Imperial College London , South Kensington Campus , London , SW7 2AZ , U.K.
ACS Synth Biol. 2018 Apr 20;7(4):1056-1066. doi: 10.1021/acssynbio.7b00435. Epub 2018 Apr 2.
Cyanobacteria are important for fundamental studies of photosynthesis and have great biotechnological potential. In order to better study and fully exploit these organisms, the limited repertoire of genetic tools and parts must be expanded. A small number of inducible promoters have been used in cyanobacteria, allowing dynamic external control of gene expression through the addition of specific inducer molecules. However, the inducible promoters used to date suffer from various drawbacks including toxicity of inducers, leaky expression in the absence of inducer and inducer photolability, the latter being particularly relevant to cyanobacteria, which, as photoautotrophs, are grown under light. Here we introduce the rhamnose-inducible rhaBAD promoter of Escherichia coli into the model freshwater cyanobacterium Synechocystis sp. PCC 6803 and demonstrate it has superior properties to previously reported cyanobacterial inducible promoter systems, such as a non-toxic, photostable, non-metabolizable inducer, a linear response to inducer concentration and crucially no basal transcription in the absence of inducer.
蓝细菌对于光合作用的基础研究很重要,并且具有巨大的生物技术潜力。为了更好地研究和充分利用这些生物,必须扩大有限的遗传工具和元件库。少数诱导型启动子已在蓝细菌中使用,通过添加特定的诱导分子实现对基因表达的动态外部控制。然而,迄今为止使用的诱导型启动子存在各种缺点,包括诱导剂的毒性、在没有诱导剂时的渗漏表达以及诱导剂的光不稳定,后者对于作为光合自养生物在光照下生长的蓝细菌尤为重要。在这里,我们将大肠杆菌的鼠李糖诱导型rhaBAD启动子引入模式淡水蓝细菌聚球藻属PCC 6803中,并证明它具有优于先前报道的蓝细菌诱导型启动子系统的特性,例如无毒、光稳定、不可代谢的诱导剂、对诱导剂浓度呈线性响应,并且至关重要的是在没有诱导剂时没有基础转录。