Steigedal Magnus, Valla Svein
Department of Biotechnology, Norwegian University of Science and Technology (NTNU), N-7491 Trondheim, Norway.
Metab Eng. 2008 Mar;10(2):121-9. doi: 10.1016/j.ymben.2007.08.002. Epub 2007 Sep 6.
Over the last 10 years there has been an extremely fast development in the global characterization of bacteria at the genome, transcriptome, proteome and metabolome levels. To further explore and apply these complex data sets there is now a need for new biological tools that can be used to test or verify hypotheses generated on the basis of all the new information. Here, we report the integration of an expression cassette based on the Acinetobacter sp. chnB promoter and its cognate positive regulator chnR gene into a replicon derived from the broad-host-range plasmid RK2. Cyclohexanone was found to be the most efficient inducer of this system in Escherichia coli, using firefly luciferase as a reporter. To explore the potential of the system in another species, we show that the system can be used in combination with another similar expression cassette (Pm/xylS) to control the monomer composition of the industrially widely used exopolysaccharide alginate, produced by Pseudomonas fluorescens.
在过去10年里,细菌在基因组、转录组、蛋白质组和代谢组水平上的全球特征研究取得了极其快速的发展。为了进一步探索和应用这些复杂的数据集,现在需要新的生物学工具,用于检验或验证基于所有这些新信息所产生的假设。在此,我们报告了将基于不动杆菌属sp. chnB启动子及其同源正调控基因chnR的表达盒整合到源自广宿主范围质粒RK2的复制子中。以萤火虫荧光素酶作为报告基因,发现环己酮是该系统在大肠杆菌中最有效的诱导剂。为了探索该系统在另一个物种中的潜力,我们表明该系统可与另一个类似的表达盒(Pm/xylS)联合使用,以控制荧光假单胞菌产生的、工业上广泛使用的胞外多糖藻酸盐的单体组成。