Lee Sung Kuk, Keasling Jay D
Department of Chemical Engineering, University of California, Berkeley, 94720, USA.
Gene. 2006 Aug 1;377:6-11. doi: 10.1016/j.gene.2006.02.013. Epub 2006 Apr 17.
The expression and regulatory properties of a propionate-regulated overexpression system (Salmonella enterica prpBCDE promoter (P(prpB)) and positive regulator (prpR)) were evaluated in wild-type S. enterica serovar Typhimurium TR6583 and prpB(-) or prpD(-) versions of this strain and compared with the arabinose-regulated T7 expression system. The wild-type strain showed low expression in the absence of propionate and high expression in the presence of propionate under all growth conditions. In 96-well plates and culture tubes, the wild-type strain exhibited a long delay before full induction; the time delay was significantly shorter in shake flasks. The prpD(-) strain exhibited low expression in the presence of glucose, highly regulatable expression over a wide range of propionate concentrations, and, in contrast to the wild-type strain, fast induction to full expression under all growth conditions. In contrast, the prpB(-) strain showed very high background expression in both culture tubes and shake flasks.
在野生型肠炎沙门氏菌鼠伤寒血清型TR6583及其prpB(-)或prpD(-)版本菌株中评估了丙酸酯调节的过表达系统(肠炎沙门氏菌prpBCDE启动子(P(prpB))和正调控因子(prpR))的表达及调控特性,并与阿拉伯糖调节的T7表达系统进行了比较。野生型菌株在所有生长条件下,在无丙酸酯时表达水平低,在有丙酸酯时表达水平高。在96孔板和培养管中,野生型菌株在完全诱导前有很长的延迟期;在摇瓶中,延迟时间明显更短。prpD(-)菌株在有葡萄糖存在时表达水平低,在广泛的丙酸酯浓度范围内具有高度可调节的表达,并且与野生型菌株不同,在所有生长条件下都能快速诱导至完全表达。相比之下,prpB(-)菌株在培养管和摇瓶中均表现出非常高的背景表达。