Reverchon S, Nasser W, Robert-Baudouy J
Laboratoire de Génétique Moléculaire des Microorganismes, Institut National des Sciences Appliquées, Villeurbanne, France.
Mol Microbiol. 1991 Sep;5(9):2203-16. doi: 10.1111/j.1365-2958.1991.tb02150.x.
Erwinia chrysanthemi is a phytopathogenic enterobacterium able to degrade the pectic fraction of plant cell walls. The kdgR negative regulatory gene controls all the genes involved in pectin catabolism, including the pel genes encoding pectate lyases. The E. chrysanthemi kdgR regulatory gene was subcloned in Escherichia coli where it was shown to be functional, since it repressed the expression of a pelE::uidA fusion. The nucleotide sequence of kdgR contained an open reading frame of 918bp preceded by classical transcriptional initiation signals. KdgR shows similarity to two other regulatory proteins, namely GylR, encoding an activator protein of the glycerol operon in Streptomyces coelicolor, and IclR, encoding a repressor of the acetate operon in Salmonella typhimurium and in Escherichia coli. Previously, comparison of regulatory regions of several genes controlled by kdgR revealed the existence of a conserved region which was proposed as a KdgR-binding site. The 25 bp oligonucleotide AAAAAAGAAACATTGTTTCATTTGT corresponding to this consensus was substituted to the lac operator, at the beginning of transcription of the lacZ gene. This construct functioned as an operator for binding of the KdgR protein in vivo.
菊欧文氏菌是一种能够降解植物细胞壁果胶部分的植物病原性肠杆菌。kdgR负调控基因控制着所有参与果胶分解代谢的基因,包括编码果胶酸裂解酶的pel基因。菊欧文氏菌的kdgR调控基因被亚克隆到大肠杆菌中,结果显示它具有功能,因为它抑制了pelE::uidA融合基因的表达。kdgR的核苷酸序列包含一个918bp的开放阅读框,其前面是典型的转录起始信号。KdgR与另外两种调控蛋白相似,即编码天蓝色链霉菌中甘油操纵子激活蛋白的GylR,以及编码鼠伤寒沙门氏菌和大肠杆菌中乙酸盐操纵子阻遏蛋白的IclR。此前,对kdgR控制的几个基因的调控区域进行比较,发现存在一个保守区域,该区域被认为是KdgR结合位点。与该共有序列对应的25bp寡核苷酸AAAAAAGAAACATTGTTTCATTTGT在lacZ基因转录开始时被替换到lac操纵子上。该构建体在体内起到了KdgR蛋白结合的操纵子的作用。