Mimura S, Rao U, Yoshino S, Kato M, Tsukagoshi N
Department of Biological Mechanisms and Functions, Graduate School of Bioagricultural Sciences, Nagoya University, Japan.
Microbiol Res. 1999 Jan;153(4):369-76. doi: 10.1016/S0944-5013(99)80052-4.
Regulation of the Chaetomium gracile xylanase A gene (cgxA) was investigated using Aspergillus nidulans as an intermediate host. Deletion of a 185 bp DNA fragment from its promoter region led to higher levels of the cgxA gene expression, indicating that the 185 bp DNA fragment contains an element involved in repression of the gene. A nuclear extract was assayed for proteins which bind to the 185 bp DNA fragment. A protein designated AnRP bound sequence specifically to the DNA fragment. The minimum sequence required for AnRP binding, 5'TTGACAAAT-3', was determined by means of gel mobility shift assays with various double-stranded oligonucleotides. Furthermore, this sequence repressed the expression of the cgxA gene when inserted at the 5' end of the cgxA gene on pXAH, which was deleted for the repressive element from the promoter region.
利用构巢曲霉作为中间宿主研究了纤细毛壳木聚糖酶A基因(cgxA)的调控。从其启动子区域缺失一个185 bp的DNA片段导致cgxA基因表达水平升高,表明该185 bp的DNA片段包含一个参与基因抑制的元件。对一种核提取物进行了与185 bp DNA片段结合的蛋白质检测。一种名为AnRP的蛋白质与该DNA片段具有序列特异性结合。通过使用各种双链寡核苷酸进行凝胶迁移率变动分析,确定了AnRP结合所需的最小序列为5'TTGACAAAT-3'。此外,当该序列插入到pXAH上cgxA基因的5'端时,抑制了cgxA基因的表达,pXAH的启动子区域已缺失抑制元件。