Abstract:
Synechococcus elongatus PCC 11801 is a fast-growing cyanobacterium, exhibiting high tolerance to environmental stresses. We have earlier characterized its genome and analysed its transcriptome and proteome. However, to deploy it as a potential cell factory, it is necessary to expand its synthetic biology toolbox, including promoter elements and ribosome binding sites (RBSs). Here, based on the global transcriptome analysis, 48 native promoters of the genes with high transcript count were characterized using a fluorescent reporter system. The promoters PcpcB, PpsbA1, and P11770 exhibited consistently high fluorescence under all the cultivation conditions. Similarly, from the genome data and proteome analysis, 534 operons were identified. Fifteen intergenic regions exhibiting higher protein expression from the downstream gene were systematically characterized for identifying RBSs, using an operon construct comprising fluorescent protein genes eyfp and mTurq under PcpcB (PcpcB:eyfp:RBS:mTurq:TrrnB). Overall, the work presents promoter and RBS sequence libraries, with varying strengths, to expedite bioengineering of PCC 11801.
摘要:
长毛蓝藻PCC11801是一种快速生长的蓝细菌,对环境压力表现出高耐受性。我们较早地对其基因组进行了表征,并分析了其转录组和蛋白质组。然而,将其部署为潜在的细胞工厂,有必要扩展其合成生物学工具箱,包括启动子元件和核糖体结合位点(RBS)。这里,基于全球转录组分析,使用荧光报告系统表征具有高转录物计数的基因的48个天然启动子。启动子PcpcB,PpsbA1和P11770在所有培养条件下都表现出一致的高荧光。同样,从基因组数据和蛋白质组分析,鉴定出534个操纵子。系统表征了15个来自下游基因的较高蛋白质表达的基因间区域,以鉴定RBS,在PcpcB下使用包含荧光蛋白基因eyfp和mTurq的操纵子构建体(PcpcB:eyfp:RBS:mTurq:TrrnB)。总的来说,该工作提出了启动子和RBS序列库,不同的力量,加快PCC11801的生物工程。