Application of the Luminescent luxCDABE Gene for the Rapid Screening of Antibacterial Substances Targeting Pseudomonas aeruginosa

发光luxCDABE基因在快速筛选靶向铜绿假单胞菌的抗菌物质中的应用

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Abstract

Pseudomonas aeruginosa (P. aeruginosa) is a typical Gram-negative bacterium that can cause the spoilage of catered food products. Using a luminescent reporter gene (luxCDABE), this study sought to construct a cell-based biosensor (PAO1-CE) to rapidly screen antibacterial substances against P. aeruginosa. A total of six antibiotics belonging to five categories were used as the model test substances. The results of the bioluminescence detection method were verified using traditional antibacterial research assessments. The correlation coefficient of the regression equation fitting the data generated using this method was greater than 0.98, supporting the credibility of this approach. Additionally, the EC(50) of each of the antibiotics assessed in this study was lower than the 1/2 MIC determined by conventional means. All six of the antibiotics caused varying degrees of damage to the cell membrane and cell wall of P. aeruginosa. Importantly, this novel method helped shorten the time necessary for active-compound detection and could be used for high-throughput detection, which would also help improve the detection efficiency. The application of this method towards the discovery of novel antibacterial compounds targeting P. aeruginosa holds substantial promise for greatly improving the efficiency of compound discovery.

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