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编码水生拉恩氏菌果聚糖蔗糖酶的lsrA基因生长阶段依赖性表达的分子特征分析

Molecular characterization of the growth phase-dependent expression of the lsrA gene, encoding levansucrase of Rahnella aquatilis.

作者信息

Seo Jeong-Woo, Jang Ki-Hyo, Kang Soon Ah, Song Ki-Bang, Jang Eun Kyung, Park Buem-Seek, Kim Chul Ho, Rhee Sang-Ki

机构信息

Korea Research Institute of Bioscience and Biotechnology (KRIBB), 51 Oun-dong, Yusong, Daejon 305-333, Korea.

出版信息

J Bacteriol. 2002 Nov;184(21):5862-70. doi: 10.1128/JB.184.21.5862-5870.2002.

Abstract

Expression of the lsrA gene from Rahnella aquatilis, encoding levansucrase, is tightly regulated by the growth phase of the host cell; low-level expression was observed in the early phase of cell growth, but expression was significantly stimulated in the late phase. Northern blot analysis revealed that regulation occurred at the level of transcription. The promoter region was identified by primer extension analysis. Two opposite genetic elements that participate in the regulation of lsrA expression were identified upstream of the lsrA gene: the lsrS gene and the lsrR region. The lsrS gene encodes a protein consisting of 70 amino acid residues (M(r), 8,075), which positively activated lsrA expression approximately 20-fold in a growth phase-dependent fashion. The cis-acting lsrR region, which repressed lsrA expression about 10-fold, was further narrowed to two DNA regions by deletion analysis. The concerted action of two opposite regulatory functions resulted in the growth phase-dependent activation of gene expression in Escherichia coli independent of the stationary sigma factor sigma(S).

摘要

编码果聚糖蔗糖酶的水生拉恩菌lsrA基因的表达受宿主细胞生长阶段的严格调控;在细胞生长早期观察到低水平表达,但在后期表达受到显著刺激。Northern印迹分析表明,调控发生在转录水平。通过引物延伸分析鉴定了启动子区域。在lsrA基因上游鉴定出两个参与lsrA表达调控的相反遗传元件:lsrS基因和lsrR区域。lsrS基因编码一种由70个氨基酸残基组成的蛋白质(分子量为8075),它以生长阶段依赖的方式使lsrA表达正向激活约20倍。通过缺失分析,将抑制lsrA表达约10倍的顺式作用lsrR区域进一步缩小到两个DNA区域。两种相反调控功能的协同作用导致大肠杆菌中基因表达的生长阶段依赖性激活,且不依赖于静止期σ因子σ(S)。

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