铜绿假单胞菌铁摄取调节子Fur诱导表达突变株构建及表型分析

铁摄取调节子 (Ferric uptake regulator,Fur) 是细菌控制细胞内铁平衡的一类重要的调节子。铜绿假单胞菌Pseudomonas aeruginosa的fur为必需基因,不能直接敲除。文中通过构建诱导型缺失突变株Δfur/attB::PBAD-fur,来研究该基因对铜绿假单胞菌的生长、生物被膜形成、运动能力和抗氧应激能力等方面的影响。结果表明,当Fur低表达时,铜绿假单胞菌在高铁和低铁环境中出现了生长阻滞的现象;低表达Fur的铜绿假单胞菌抵抗H2O2的能力降低,形成生物被膜的能力减弱,游动、颤动 (Twitching) 和丛集 (Swarming) 运动能力也出现了减弱的现象。Fur的表达直接影响铜绿假单胞菌荧光嗜铁素的产量。在铜绿假单胞菌体内表达来自格瑞菲斯瓦尔德磁螺菌Fur超级家族中的蛋白,可以部分恢复铜绿假单胞菌荧光嗜铁素的产量。由此说明,Fur对铜绿假单胞菌的生长、生物被膜形成、抗氧应激能力和运动能力方面都起着至关重要的作用。本研究为铜绿假单胞菌的防治提供理论指导。     

Effects of localised,low-voltage pulsed electric fields on the development and inhibition of Pseudomonas aeruginosa biofilms

Abstract

This work describes the use of low-voltage (0.5 – 5 V) pulsed electric fields to prevent Pseudomonas aeruginosa biofilm development. Interdigitated electrodes (IDEs) with 29-μm spacing between 22-μm-wide electrodes, were used as a platform where the effect of localised, high-strength electric fields could be tested. Alternating current, square-wave pulses were applied to the IDEs in 1 sec intervals. A two-level, three-variable factorial design experiment was used to detect the effects of applied voltage, frequency, and pulse duty ratio (i.e. percentage of pulsing time over one cycle) on the inhibition of biofilm formation. The observations indicated that a pulse configuration of 1% duty ratio, 5 V, and 200 Hz frequency reduced the area of the electrodes covered by biofilm by 50%. In general, the application of low-duty ratio pulses had a positive effect on preventing biofouling. Comparatively, frequency and applied voltage were observed to have less influence on biofouling.