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【目的】研究铜绿假单胞菌PAO1 PA2580基因的功能。【方法】构建了PA2580的敲除突变体及突变体互补体,通过最小抑制浓度测定、基因启动子活性检测、蛋白体外表达纯化等方法,对PA2580基因的功能进行了深入的研究。【结果】PA2580突变体对羧苄青霉素、氯霉素、环丙沙星的敏感性增强。PA2580基因的表达还受到不同种类的低于抑制浓度的抗生素的调节。PA2580蛋白产物以NADPH为电子供体,能够高效还原多种醌类物质。此外,PA2580突变体对过氧化氢敏感性增加,过氧化氢酶编码基因在PA2580突变体中的表达降低,表明PA2580与铜绿假单胞菌氧化压力耐受性相关。【结论】PA2580产物是NADPH-醌类的还原酶,其功能与铜绿假单胞菌对环境压力的耐受密切相关。
【Objective】 To study the function of PAO1 PA2580 gene in Pseudomonas aeruginosa. 【Method】 PA2580 knockout mutants and mutants were constructed. The function of PA2580 gene was studied in detail by the methods of minimal inhibitory concentration assay, detection of gene promoter activity and in vitro expression and purification of PA2580. 【Result】 PA2580 mutants were more sensitive to carbenicillin, chloramphenicol and ciprofloxacin. The expression of the PA2580 gene is also regulated by different kinds of antibiotics at concentrations below the inhibitory concentration. PA2580 protein products with NADPH as electron donor, can efficiently restore a variety of quinones. In addition, the sensitivity of PA2580 mutant to hydrogen peroxide was increased, and the expression of catalase gene in PA2580 mutant was decreased, indicating that PA2580 was associated with oxidative stress tolerance of Pseudomonas aeruginosa. 【Conclusion】 PA2580 product is a reductase of NADPH-quinones, and its function is closely related to the tolerance of Pseudomonas aeruginosa to environmental pressure.