Miksch G, Bettenworth F, Friehs K, Flaschel E, Saalbach A, Nattkemper T W
Lehrstuhl für Fermentationstechnik, Technische Fakultät, Universität Bielefeld, Germany.
Appl Microbiol Biotechnol. 2006 Mar;70(2):229-36. doi: 10.1007/s00253-005-0060-4. Epub 2005 Jul 13.
To develop a rapid reporter system for the screening of stationary-phase promoters in Escherichia coli, the expression pattern of the green fluorescent protein (GFP) during bacterial cultivation was compared with that of the commonly used beta-galactosidase. Using GFP with enhanced fluorescence, the expression pattern of both reporter systems GFP and beta-galactosidase were similar and showed a typical induction of gene activity of the reporter genes, i.e. increase of expression at the transition from exponential to stationary phase. The expression was affected by the culture medium, i.e. in contrast to the complex medium (LB medium), the stationary-phase specific induction was only observed in synthetic medium (M9) when amino acids were added, whereas there was generally no induction in MOPS medium. To develop a rapid screening method on agar plates for stationary-phase promoters, a photographic approach was used, continued with computational image treatment. A screening method is presented which enables an on-line monitoring of gene activity.
为开发一种用于筛选大肠杆菌中稳定期启动子的快速报告系统，将细菌培养过程中绿色荧光蛋白（GFP）的表达模式与常用的β-半乳糖苷酶的表达模式进行了比较。使用具有增强荧光的GFP，GFP和β-半乳糖苷酶这两种报告系统的表达模式相似，并显示出报告基因典型的基因活性诱导，即在从指数期到稳定期过渡时表达增加。表达受培养基影响，即与复杂培养基（LB培养基）相比，仅在添加氨基酸的合成培养基（M9）中观察到稳定期特异性诱导，而在MOPS培养基中通常没有诱导。为开发一种在琼脂平板上快速筛选稳定期启动子的方法，采用了摄影方法，并继续进行计算图像处理。提出了一种能够在线监测基因活性的筛选方法。
