Gaffney T D, da Costa e Silva O, Yamada T, Kosuge T
Department of Plant Pathology, University of California, Davis 95616.
J Bacteriol. 1990 Oct;172(10):5593-601. doi: 10.1128/jb.172.10.5593-5601.1990.
Expression of the indoleacetic acid (iaa) operon, which contributes to the virulence of the phytopathogenic bacterium Pseudomonas syringae subsp. savastanoi, was monitored by using broad-host-range lacZ reporter gene plasmids. A combination of translational (gene) fusions and transcriptional (operon) fusions of P. syringae subsp. savastanoi sequences to lacZ allowed localization of the iaa operon promoter. RNA recovered from P. syringae subsp. savastanoi strains was mapped with iaa operon-specific probes to precisely locate the transcription initiation site. When transcripts from an iaaM::lacZ fusion in Escherichia coli were analyzed, an identical transcription initiation site was observed. The DNA sequence of the iaa operon promoter closely resembled the consensus E. coli promoter sequence. We detected an active, constitutive level of indoleacetic acid biosynthetic gene expression during bacterial growth under a variety of conditions in the absence of host plant influence.
利用广宿主范围的lacZ报告基因质粒监测了吲哚乙酸(iaa)操纵子的表达,该操纵子有助于植物致病细菌丁香假单胞菌 savastanoi 亚种的毒力。丁香假单胞菌 savastanoi 亚种序列与lacZ的翻译(基因)融合和转录(操纵子)融合相结合,使得iaa操纵子启动子得以定位。从丁香假单胞菌 savastanoi 亚种菌株中回收的RNA用iaa操纵子特异性探针进行定位,以精确确定转录起始位点。当分析大肠杆菌中iaaM::lacZ融合的转录本时,观察到了相同的转录起始位点。iaa操纵子启动子的DNA序列与大肠杆菌启动子共有序列非常相似。我们发现在没有宿主植物影响的各种条件下细菌生长过程中,吲哚乙酸生物合成基因表达处于活跃的组成型水平。