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Microarray Analysis of Pseudomonas aeruginosa Quorum-Sensing Regulons: Effects of Growth Phase and Environment†

机译:铜绿假单胞菌群体敏感调节子的微阵列分析:生长期和环境的影响†

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摘要

Bacterial communication via quorum sensing (QS) has been reported to be important in the production of virulence factors, antibiotic sensitivity, and biofilm development. Two QS systems, known as the las and rhl systems, have been identified previously in the opportunistic pathogen Pseudomonas aeruginosa. High-density oligonucleotide microarrays for the P. aeruginosa PAO1 genome were used to investigate global gene expression patterns modulated by QS regulons. In the initial experiments we focused on identifying las and/or rhl QS-regulated genes using a QS signal generation-deficient mutant (PAO-JP2) that was cultured with and without added exogenous autoinducers [N-(3-oxododecanoyl) homoserine lactone and N-butyryl homoserine lactone]. Conservatively, 616 genes showed statistically significant differential expression (P ≤ 0.05) in response to the exogenous autoinducers and were classified as QS regulated. A total of 244 genes were identified as being QS regulated at the mid-logarithmic phase, and 450 genes were identified as being QS regulated at the early stationary phase. Most of the previously reported QS-promoted genes were confirmed, and a large number of additional QS-promoted genes were identified. Importantly, 222 genes were identified as being QS repressed. Environmental factors, such as medium composition and oxygen availability, eliminated detection of transcripts of many genes that were identified as being QS regulated.
机译:据报道,通过群体感应(QS)进行的细菌交流在产生毒力因子,抗生素敏感性和生物膜发育方面很重要。先前已经在机会病原体铜绿假单胞菌中发现了两个称为las和rhl系统的QS系统。用于铜绿假单胞菌PAO1基因组的高密度寡核苷酸微阵列用于研究QS调节子调节的全局基因表达模式。在最初的实验中,我们专注于使用QS信号产生缺陷型突变体(PAO-JP2)鉴定las和/或rhl QS调控的基因,该突变体在有和没有添加外源自体诱导剂[N-(3-氧十二烷酰基)高丝氨酸内酯和N-丁酰基高丝氨酸内酯]。保守地,有616个基因在对外源自体诱导剂的反应中表现出统计学上显着的差异表达(P≤0.05),并被归类为QS调控。总共244个基因在对数中期被鉴定为受QS调控,而450个基因在静止早期被鉴定为被QS调控。确认了大多数先前报道的QS促进基因,并鉴定了许多其他QS促进基因。重要的是,鉴定出222个基因被QS抑制。环境因素(例如培养基组成和氧气供应量)消除了对许多被鉴定为QS调控基因的转录本的检测。

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